Expression profile of lncRNA induced by DPP4 in human aortic smooth muscle cells and calcification gene prediction

Expression profile of lncRNA induced by DPP4 in human aortic smooth muscle cells and calcification gene prediction | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Expression profile of lncRNA induced by DPP4 in human aortic smooth muscle cells and calcification gene prediction Tongjie Xu, Hao Chen, Xiongfei Xu, Xiaolei Sun, Huqiang He, Hong Zeng, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-1656423/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Arterial media calcification caused by diabetes is an important cause of cardiovascular calcification. Dipeptidyl peptidase-4 (DPP4) is associated with diabetic arterial media calcification. While, long non-coding RNA(lncRNA) is closely related to the cardiovascular diseases and calcification caused by diabetes. Experiment have shown that LncRNA can regulate DPP4 activation to modulate senescence and lncRNA ENST00000540293 may associated with human periodontal ligament cells development. our previous studies showed that DPP4 can promote the calcification of human aortic smooth muscle cells (HASMCs) by activating the ERK/NF-kB pathway, but it is unknown whether it is related to the regulation of lncRNA. Calcification of HASMCs was induced by DPP4. There was a significant difference in the expression of lncRNAs and mRNAs between normal and calcified cells detected by gene chip technology. Based on the results of microarray detection, we found that lncRNA ENST00000540293 may be involved in vascular calcification induced by DPP4 through regulating target genes. Dipeptidyl peptidase 4 Vascular Calcification RNA Chip Long non-coding RNA Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 1. Introduction Vascular calcification is a common pathological phenomenon in coronary atherosclerotic heart disease, chronic kidney disease, diabetes and other diseases 1 , Its pathological basis is an expression of osteoblast-specific genes in smooth muscle cells 2 . Previous studies have shown that vascular calcification is a very complex pathological process, which is regulated by many factors. Recent studies have found that vascular calcification is an adjustable and active biological process, which is similar to bone formation and involves the expression of some bone formation related proteins, such as alkaline phosphatase (ALP), osteocalcin (OCN), Runx2 and BMP-2 3 . Vascular calcification is common in patients with diabetes, and the important pathological change of vascular complications is intima-media calcification. Medial calcification and cardiovascular disease caused by diabetes are one of the main causes of death. Vascular calcification is closely related to the high incidence and mortality of cardiovascular disease. Therefore, it is of great significance to study the treatment and intervention of diabetic vascular calcification. Dipeptidyl peptidase 4 (DPP4) is a highly glycosylated type II transmembrane protein composed of 766 amino acids, which belongs to the family of serine proteases. One of its catalytic substrates is incretin. The treatment of type 2 diabetes with DPP4 inhibitors (Letins) establishes the role of incretin in blood glucose control. Low molecular weight DPP4 inhibitors exert their hypoglycemic effect by promoting glucose-dependent insulinotropic polypeptide (GIP) and inhibiting the degradation of glucagon-like peptide 1 (GLP-1). DPP4 is widely expressed in a variety of cells, such as endothelial cells and immune cells. DPP4 also plays an important role in the regulation of cardiovascular function 4 . Meanwhile, DPP4 not only plays an important role in reducing blood sugar, but also has unique functions in other aspects 5 . Nina Wronkowitz et al demonstrated that soluble DPP4 induces inflammation and proliferation of human smooth muscle cells via protease-activated receptor 2 6 . Yu et al demonstrated that soluble DPP4 can induce oxidative stress in human umbilical vein endothelial cells through mannose 6-phosphate/insulin-like growth factor receptor II 7 . A.M.Lambeir et al found that the expression of DPP4 in visceral tissues of obese and diabetic patients was higher than normal people 8 . Studies have shown that DPP4 and its inhibitors are closely related to vascular calcification 9 . DPP4 may lead to vascular calcification 10 . While,DPP4 inhibitors may lowered the levels of fasting serum glucose, triglyceride (TG),TNF-α and decreased the calcium deposits 11 , and also attenuated vascular calcification and osteogenic trans-differentiation in vascular smooth muscle cells via multiple steps, such as: downregulation of PiT-1 expression and suppression of reactive oxygen species generation, phospho-PI3K/AKT, and the Wnt signaling pathway 12 .The above data suggest that diabetes and its concomitant diseases may be closely related to the regulation of DPP4. Long non-coding RNA (lncRNA) is a functional RNA molecule with a length of more than 200bp in non-coding RNA 13 . LncRNA is located in the cytoplasm or nucleus. They regulate gene expression through epigenetic, transcriptional and post-transcriptional regulation in the form of RNA, but lack the ability of protein coding. It has been found that lncRNA plays an important role in regulating cell growth, development, distribution and targeted differentiation 14 . Intergenic lncRNA(lincRNA) and antisense lncRNA are one of the hotspots in the current research. LincRNA ENST00000540293 is an intergenic noncoding RNA located on chromosome 11 and contains 1000 bases, and may associated with human periodontal ligament cells development 15 . Our RNA microarray results show that the lncRNA ENST00000540293 nearby gene is NM-002421, and the protein encoded by NM-002421 is matrix metalloproteinase 1 (MMP-1) ( Table 1 ) 16 . Previous studies have shown that matrix metalloproteinases are closely related to the occurrence and development of vascular calcification 17,18 . Our preliminary experiments in vitro experiments showed that DPP4 can promote the calcification of HASMCs by activating the ERK/NF-kB pathway, and the DPP4 inhibitors (sitagliptin) can inhibit ERK1/2 and NF-kB signaling pathway so that inhibits aortic vascular calcification in type 2 diabetic mice also confirmed in vivo. Although DPP4 is closely related to cardiovascular disease, the exact mechanism of DPP4 promoting HASMCs calcification is not completely clear 11 . Therefore, in this study, HASMCs were cultured with DPP4 to establish calcification model, so as to study the mechanism of DPP4 promoting osteogenic differentiation of vascular smooth muscle cells. 2. Materials And Methods 2.1 The culture of HASMCs The primary human aortic vascular smooth muscle cell (HASMCs,no.6110) and culture medium (no.1101)were obtained from Science Cell Research-Laboratory (USA). SMCM medium was composed of 500ml basic medium, 10ml fetal bovine serum (FBS, no.0010), 5ml smooth muscle cell growth factor (SMCGS, no.1152) and 5ml penicillin / streptomycin (Pmax S, no.0503).The 3rd to 8th generation cells were selected for the experiment and cultured in an incubator at 37℃with 5% humidified CO2. 2.2 Determination of intracellular calcium concentration The cell intracellular calcium concentration was determined using a spectrophotometer at 570 nm. Transferring 800ul of the buffer to another 1 ml cuvette and adding 100 ul of the reaction solution by pipette. They were evenly mixed and incubated at room temperature for 5 minutes. Taking 100 μl mixture liquid in 96-well plates with 3-6 multiple holes in each group. The standard curve was plotted with the optical density (OD) on the y axis and the standard calcium concentration (mmol/l) on the x axis and get the calcium concentration. 2.3 Western blotting The levels of the all target proteins was measured by the fluorescence imaging system Image Studio, and the expression level of protein was quantified by measuring the greyscale of bands and normalized to β-actin. Briefly,the concentration of protein was detected using a BCA kit (Beyotime Biotechnology, Shanghai,China). Then, Approximately 20μg of protein was loaded onto a 10% SDS-PAGE gel and blotted onto PVDF membrane.After blocking with 5% BSA, the membranes were incubated with primary antibodies at 4°C overnight. The membranes were washed thrice by TBST and incubated with secondary antibodies for 1 h at room temperature.Such as Osteoprotegerin (OPG), Bone Morphogenetic Protein 2 (BMP-2), Osteopontin (OPN), Runt Related Transcription Factor 2 (RUNX2), α Smooth Muscle actin (α-SMA), Matrix Metalloproteinase 1 (MMP-1) were detected by western blotting with their respective antibodies(anti-OPG,OPN,BMP-2 ,MMP-1 all from Abcam; anti-α-SMA from R&D Systems;anti-RUNX2 from Cell Signaling Technology). 2.4 Alizarin red staining for Calcification Detection Cell calcification nodules were detected by Alizarin red kit（Sigma USA)）.Control group and experimental group cells were inoculated on 12-well plates. Cleaning solution was added to each well to clean the growing cell surface, fixed solution was added and incubated at room temperature for 15 minutes, then staining solution was added and incubated at room temperature for 5 minutes or until orange-red was visible. The staining solution was removed and dried in air at room temperature for about 10 minutes. Then transparent solution was added and immediately observed under light microscope: the positive cells of calcium deposition were orange-red. 2.5 F-actin staining The HASMCs (2×10 5 cells/well) were inoculated on a 6-well plate overnight and then treated with the tested compound for 72 hours. Cells were washed with PBS and fixed with 4% Formaldehyde, Permeabilized in 0.2% Triton X-100 at room temperature for 5 minutes, then stained with DAPI (Beyotime,USA) and Rhodamine Phalloidin (1:200,Sigma,MO,USA) to mark the cyteblast and cytoskeleton respectively. Pictures were acquired by Photometrics CoolSNAPHQ2 CCD camera on the Olympus IX71-Applied Precision Delta Vision restoration microscope (Applied Precision, USA) and deconvolved using Delta Vision algorithms (Applied Precision, USA). 2.6 Detection of intracellular ALP activity For ALP activity measurement,the calcified HASMCs were solubilized with RIPA lysis buffer (Beyotime Biotechnology, Shanghai,China). After centrifugation, the supernatants were examined with the ALP activity kit (Beyotime Biotechnology,Shanghai,China) .The optical density (OD) value of each group was determined by 450nm wavelength. The standard curve was made and the corresponding ALP activity of each group was calculated. 2.7 RNA extraction and real-time quantitative polymerase chain reaction. Total RNA was extracted using TRI zol reagent (Invitrogen;Thermo Fisher Scientific, Inc). RNA quantity and quality were measured by Nano Drop ND-1000. RNA integrity was assessed by standard denaturing agarose gel electrophoresis. Based on the predicted consequences, we selected the target linRNA with the most expression change between the control group and DPP4 intervened group and designed PCR primers ( Table 2 ). RT‑qPCR was performed using SYBR Green Real‑Time PCR Master mix (Invitrogen;Thermo Fisher Scientific, Inc),and was run at 95˚C for 5 min, followed by 40 cycles at 95˚Cfor 10 sec, 65˚C for 20 sec and 72˚C for 30 sec. The characteristics of the linRNAs and mRNAs are presented in Table 1 and Table 3 . Data were normalized to β-actin and the relative level of gene expression was calculated using the 2 -ΔΔCt method. 2.8 Microarray Arraystar Human LncRNA Microarray V4.0 is designed for the global profiling of human lncRNAs and protein-coding transcripts, which is updated from the previous Microarray V3.0. About 40,173 lncRNAs and 20,730 coding transcripts can be detected by our third-generation lncRNA microarray. 2.9 RNA labeling and array hybridization Sample labeling and array hybridization were performed according to the Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technology) with minor modifications. Briefly, mRNA was purified from total RNA after removal of rRNA (mRNA-ONLY Eukaryotic mRNA Isolation Kit, Epicentre). Then, each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 3' bias utilizing a random priming method (Arraystar Flash RNA Labeling Kit, Arraystar). The labeled cRNAs were purified by RNeasy Mini Kit (Qiagen). The concentration and specific activity of the labeled cRNAs (Pmol Cy3/μg cRNA) were measured by NanoDrop ND-1000. 1 μg of each labeled cRNA was fragmented by adding 5 μl 10 × Blocking Agent and 1 μl of 25 × Fragmentation Buffer, then heated the mixture at 60°C for 30 min, finally 25 μl 2 × GE Hybridization buffer was added to dilute the labeled cRNA. 50 μl of hybridization solution was dispensed into the gasket slide and assembled to the LncRNA expression microarray slide. The slides were incubated for 17 hours at 65°C in an Agilent Hybridization Oven. The hybridized arrays were washed, fixed and scanned with using the Agilent DNA Microarray Scanner (part number G2505C). 2.10 LncRNA transient transfection. VSMCs were interference with 50 nm lncRNA ENST00000540293 Smart Silencer (siRNA ) and negative control (NC-siRNA) using Smart Silencer NC at a density of 4 x10 5 cells for 48 hours, according to the manufacturer's protocol 19 . The inhibitory efficiency was tested by real-time qPCR, and the effective siRNA was picked for the further experiments. The lncRNA ENST00000540293 siRNA and NC-siRNA were synthesized by RiboBioCo (Guangzhou, China). Statistical analysis. Data are presented as the mean ± standard deviation (n=3) and were analyzed using a Student's t-test. All statistical analysis was performed with SPSS 20.0 (IBM Corp, Armonk, NY, USA). P value < 0.05 was accepted as significant. 3. Results 1 .Effect of DPP4 on HASMCs calcification protein The 5rd to 7th generation HASMCs were cultured with 200ng/ml DPP4 for 7 days. The total cell protein was extracted and the expression of OPG, OPN, BMP-2 and RUNX2 was detected by western blotting. The expression of OPG, OPN, BMP-2 and RUNX2 protein in DPP4 group was significantly higher than control group ( Fig.1A ). The results confirm that DPP4 can induce HASMCs calcification. 2 .Calcification of HASMCs induced by DPP4 After cultured with 200ng/ml DPP4 for 14 days, the cells in the control group showed typical multi-layer smooth muscle cells, and the alizarin red staining was negative, while the orange red calcified nodules in the DPP4 group were significantly more than those in the control group.The alizarin red staining was strongly positive ( Fig.1Ban Fig.1D ),and calcium content was measured( Fig.1C) , which further confirmed that DPP4 could induce HASMCs calcification. 3.Expression profile of lncRNA and mRNA in HASMCs after DPP4 intervention After adding 200ng/ml DPP4 to HASMCs, the total RNA was extracted to detect its quality, concentration and integrity.Differentially expressed lncRNAs and mRNAs with statistical significance between the two groups were identified through P-value/FDR filtering. Differentially expressed lncRNAs and mRNAs between the two samples were identified through Fold Change filtering( Fig .2A and Fig .2B ). 4.DPP4 inhibits the expression of lncRNA ENST00000540293 and MMP-1 mRNA in HASMCs The differentially expressed lncRNAs or mRNAs between the two groups were screened by P-value/FDR, and further screened by Fold Change. The results suggest that DPP4 can inhibit the expression of lncRNA ENST00000540293 and MMP-1 mRNA ( Table 1 and Table3 ). To confirm the reliability and validity of the microarray data. LncRNAs and mRNAs chip screening results were verified by RT-qPCR. 5.Effect of inhibiting the expression of HASMCs lncRNA ENST00000540293 on calcification proteins. In order to clarify the relationship between inhibition of lncRNA ENST00000540293 expression and calcification-related protein expression, HASMCs was divided into control group, transfection control group(include Transfection Reagent only), siRNA control group and siRNA group and cultured for 48 hours. The expression of calcification-related proteins BMP-2, OPG, OPN and RUNX2 was detected by Western blot. The results showed that the expression of calcitonin in siRNA group was significantly higher than the other groups ( Fig.3 ). It was proved that inhibiting the expression of lncRNA ENST00000540293 could promote the expression of HASMCs calcitonin. 6. Morphological analysis and calcium content determination of calcification of HASMCs after inhibition of lncRNA ENST00000540293 expression. In order to further analyze the cell morphology, intracellular calcium content and the activity of alkaline phosphatase (ALP) after inhibiting the expression of lncRNA ENST00000540293 in HASMCs. HASMCs were cultured for 72 hours and stained with alizarin red. The intracellular calcium content, activity of ALP of each group was determined. The results showed : the area of calcified nodules and orange calcium nodules increased significantly in SiRNA group( Fig.4A and Fig.4B ),the activity of ALP and intracellular calcium content in SiRNA group increased significantly (***P<0.001)( Fig. 4C and Fig.4D ). Previous studies have proved that there is a positive correlation between intracellular ALP activity, intracellular calcium content and HASMCs calcification. The results showed that inhibition of epidermis of lncRNA ENST00000540293 could promote the calcification of HASMCs. 7.Inhibition of lncRNA ENST 00000540293 expression promotes phenotypic transformation of HASMCs. The mechanism of differentiation of HASMCs into osteoblast-like cells is similar to that of bone formation 2 0 .In order to further verify that inhibiting the expression of lncRNA ENST00000540293 can promote the differentiation of VSMCs into osteoblasts, we detected the expression of α-SMA by Western blot( Fig.5A ). The expression of α-SMA protein in siRNA group was significantly lower than other groups (**P <0.01). At the same time, the results of F-actin immunofluorescence staining were observed by laser confocal microscope( Fig.5B ). F-actin (red) fluorescence intensity, myofilament expression and aggregation in siRNA group were significantly decreased, further indicate that HASMCs had a tendency to transform into osteoblasts. In short, it can be speculated that after lncRNA ENST00000540293 silencing, the phenotype of HASMCs changes from contractile type to synthetic secretory type, and finally into osteoblast-like cells. 8. Silencing lncRNA ENST00000540293 promoted the expression of MMP-1 and ERK 1/2 in HASMCs. Studies have shown that DPP4 regulate the expression of MMP-1 protein through ERK1/2 signal pathway 2 1 .And lncRNA may play a role by regulating coding genes near transcription or other signal way 2 2 . Our previous gene chip microarray subarray analysis showed that there were many differentially expressed lncRNAs near mRNAs related to tissue development, in which (Nearby Gene), a gene attached to lncRNA ENST00000540293, was a protein encoded by NM-002421, which was matrix metalloproteinase-1 ( Table 1 ). It was found that after inhibiting the expression of lncRNA ENST00000540293, MMP-1 protein and ERK 1/2 phosphorylate in siRNA group was significantly higher than the other groups detected by Western blot( Fig.6A and Fig.6B ),and the difference was statistically significant (**p<0.01,***p<0.001). Discussion Vascular calcification is a pathological process in which HASMCs change from contractile type to synthetic secretory type and finally to osteoblast-like phenotype under various pathological actions, which mediates the abnormal deposition of calcium in the vascular wall. Vascular calcification exists widely in diabetes, atherosclerosis, hypertension, chronic renal failure and other diseases, and the degree of calcification is positively correlated with the severity of the disease 2 3 ,2 4 . Previous studies have shown that the occurrence and development of vascular calcification is closely related to cell degeneration and necrosis, and it is a passive and degenerative vascular terminal lesion process, while recent studies have found that vascular calcification is a complex biological process that is controllable, active, preventable and reversible similar to bone formation 2 5 . Changes in extracellular conditions and phenotypic transformation of vascular smooth muscle cells play a key role 2 6 . At the same time, vascular calcification is similar to intramembranous osteogenesis, which usually leads to an increase in vascular hardness, a decrease in vascular compliance, and an increase in the incidence and mortality of cardiovascular events, more common in diabetes, chronic kidney disease 2 7 ,2 8 ．DPP4 also known as T cell surface antigen CD26, containing 110kDa glycoprotein, is a serine protease widely expressed on the surface of all kinds of cells. As a new type of fat factor, it can decompose and metabolize glucagon-like peptide-1 (GLP-1) to regulate blood sugar, and also plays an important role in the pathogenesis of diabetes and cardiovascular disease 29 ,3 0 . At the same time, it has been found that DPP4 as an agonist of protease activated receptor 2 (PAR2), can directly activate the cascade of MAPK and NF-kB signals involved in PAR2, and induce the proliferation and inflammation of HASMCs 6 . Long non-coding RNA is a functional RNA transcribed by RNA polymerase II. It is located in the nucleus or cytoplasm. Due to the lack of the ability to encode proteins, it was once considered to have no biological function. At present, it is believed that it plays a similar role in the regulation of gene expression. Such as chromosome silencing, chromatin modification, transcriptional activation, transcriptional interference, nuclear transport and other important regulatory processes 3 1 -3 5 . Among them, lncRNA ENST00000540293 is differentially expressed in human dental follicle cells and periodontal ligament cells, and is closely related to the osteogenic differentiation of human periodontal ligament cells 15,3 6 . In this experiment, the calcification of HASMCs induced by DPP4 was used to screen out lncRNAs and mRNAs with obvious differences. The results showed that there were 25 differentially expressed inter-gene lncRNA (8 up-regulated and 17 down-regulated), and the most down-regulated inter-gene lncRNA was ENST00000540293, while lincRNA ENST00000540293 nearby gene was NM-002421, which encoded matrix metalloproteinase-1 (MMP-1). MMP-1 as the prototype of all interstitial collagenases, is also known as collagenase-1, fibroblast collagenase and interstitial collagenase 3 7 . It is closely related to the regulation of cell microenvironment, and can change cell function to regulate cell differentiation, such as osteogenic differentiation of human periodontal ligament cells and calcification of HASMCs 3 8 -4 1 . In order to further determine whether the calcification of HASMCs promoted by DPP4 is related to the regulation of lncRNA ENST00000540293. We constructed smart silencer transducers silencing lncRNA ENST00000540293 expression and infected HASMCs, to detect the changes of Runx2, BMP-2, OPG and OPN proteins. The expression of calcification proteins was significantly increased and alizarin red staining also showed a significant increase in calcified nodules after lncRNA ENST0000054029 silenced. The determination of intracellular calcium content and alkaline phosphatase activity showed the same results.Also,α-SMA which as the surface marker protein of vascular smooth muscle cells also decreased significantly after lncRNA ENST0000054029 gene knockdown.And the immunofluorescence results also showed that the fluorescence intensity of F-actin, myofilament expression and aggregation decreased significantly. It is suggested that DPP4 can induce the decrease of contractile phenotype of HASMCs, which further confirms the tendency of HASMCs to osteoblast. DPP4 is an enzyme that targets the incretin hormone GLP-1 for degradation,is also an important drug target of diabetes and new adipokines, which needs to be cleaved, exfoliated and released from the cell membrane to regulate the circulatory system 4 2 . MMP-1, MMP-2 and MMP-14 and MMP-9 are involved in the cleavage and shedding of DPP4,and the increased expression of MMP-1 can promote the shedding of DPP4 on the membrane of HASMCs under hypoxia 4 3 . DPP4 inhibitor Alogliptin reverses vascular remodelling by downregulating matrix metalloproteinase 1 expression through inhibition of the ERK1/2/NF-κB signal pathway 31 .The results suggest that DPP4 inhibits the expression of lncRNA ENST00000540293 and MMP-1mRNA. However, inhibiting the expression of lncRNA ENST00000540293 can promote the expression of MMP-1 protein and phosphorylation of ERK1/2 protein. Therefore, we speculate that DPP4 promotes HASMCs calcification by inhibiting lncRNA ENST00000540293, which may be related to the regulation of ERK1/2 signal pathway or MMP-1 protein. In this study, we are unable to investigate all mechanisms of DPP4 silencing lncRNA ENST00000540293 expression and activating ERK1/2 or MMP-1 and their mutual regulation. And Whether DPP4, MMP-1 and lncRNA ENST00000540293 can be used as key targets for the treatment of diabetic vascular calcification remains to be further studied. Declarations Acknowledgements Not applicable. Authors' contributions All authors contributed to the study conception and design. The first draft of the manuscript was written by Tongjien X; material preparation, data collection and analysis were performed by Hao C, Xiongfei X, Xiaolei S, Huqiang H, Hong Z, Sen S and Yong L was responsible for project administration, review, and editing. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript. Funding The study was supported by research grants from the National Natural Science Foundation of China (Fund No.81270358); the Science & Technology Department of Sichuan Province, Grant 2019JY0691. Availability of data and materials The data were obtained from the NCBI GEO database (Number :GSE189923).https://www.ncbi.nlm.nih.gov/geo/. Consent for publication. Not applicable. Competing interests The authors declare that there is no competing interests regarding the publication of this article. References Johnson, R. C., Leopold, J. A. & Loscalzo, J. Vascular calcification: pathobiological mechanisms and clinical implications. Circ Res 99 , 1044–1059, doi: 10.1161/01.RES.0000249379.55535.21 (2006). Demer, L. L. & Tintut, Y. Vascular calcification: pathobiology of a multifaceted disease. Circulation 117 , 2938–2948, doi: 10.1161/CIRCULATIONAHA.107.743161 (2008). Thompson, B. & Towler, D. A. Arterial calcification and bone physiology: role of the bone-vascular axis. Nat Rev Endocrinol 8 , 529–543, doi: 10.1038/nrendo.2012.36 (2012). Eddington, H. et al. Serum phosphate and mortality in patients with chronic kidney disease. Clin J Am Soc Nephrol 5 , 2251–2257, doi: 10.2215/CJN.00810110 (2010). Xu, J. et al. Renalase is a novel, soluble monoamine oxidase that regulates cardiac function and blood pressure. J Clin Invest 115 , 1275–1280 (2005). Wronkowitz, N. et al. Soluble DPP4 induces inflammation and proliferation of human smooth muscle cells via protease-activated receptor 2. Biochim Biophys Acta 1842 , 1613–1621, doi: 10.1016/j.bbadis.2014.06.004 (2014). Ishibashi, Y., Matsui, T., Maeda, S., Higashimoto, Y. & Yamagishi, S.-i. Advanced glycation end products evoke endothelial cell damage by stimulating soluble dipeptidyl peptidase-4 production and its interaction with mannose 6-phosphate/insulin-like growth factor II receptor. Cardiovascular diabetology 12 , 125, doi: 10.1186/1475-2840-12-125 (2013). Lambeir, A.-M., Durinx, C., Scharpé, S. & De Meester, I. Dipeptidyl-peptidase IV from bench to bedside: an update on structural properties, functions, and clinical aspects of the enzyme DPP IV. Crit Rev Clin Lab Sci 40 , 209–294 (2003). Lee, S. et al. Dipeptidyl peptidase-4 inhibition to prevent progression of calcific aortic stenosis. Heart (British Cardiac Society) 106 , 1824–1831, doi: 10.1136/heartjnl-2020-317024 (2020). Wang, Y. et al. Melatonin ameliorates aortic valve calcification via the regulation of circular RNA CircRIC3/miR-204-5p/DPP4 signaling in valvular interstitial cells. J Pineal Res 69 , e12666, doi: 10.1111/jpi.12666 (2020). Lin, C.-P. et al. Sitagliptin attenuates arterial calcification by downregulating oxidative stress-induced receptor for advanced glycation end products in LDLR knockout mice. Sci Rep 11 , 17851, doi: 10.1038/s41598-021-97361-w (2021). Choi, S.-Y. et al. Dipeptidyl peptidase-4 inhibitor gemigliptin protects against vascular calcification in an experimental chronic kidney disease and vascular smooth muscle cells. PloS one 12 , e0180393, doi: 10.1371/journal.pone.0180393 (2017). Kapranov, P. et al. RNA maps reveal new RNA classes and a possible function for pervasive transcription. Science (New York, N.Y.) 316 , 1484–1488 (2007). Guttman, M. & Rinn, J. L. Modular regulatory principles of large non-coding RNAs. Nature 482 , 339–346, doi: 10.1038/nature10887 (2012). Wu, L. et al. Comparison of long non–coding RNA expression profiles in human dental follicle cells and human periodontal ligament cells. Mol Med Rep 20 , 939–950, doi: 10.3892/mmr.2019.10308 (2019). Chen, W.-P., Yang, Z.-G., Hu, P.-F., Bao, J.-P. & Wu, L.-D. Acacetin inhibits expression of matrix metalloproteinases via a MAPK-dependent mechanism in fibroblast-like synoviocytes. J Cell Mol Med 19 , 1910–1915, doi: 10.1111/jcmm.12564 (2015). Purroy, A. et al. Matrix metalloproteinase-10 deficiency delays atherosclerosis progression and plaque calcification. Atherosclerosis 278 , 124–134, doi: 10.1016/j.atherosclerosis.2018.09.022 (2018). Provenzano, M. et al. The Association of Matrix Metalloproteinases with Chronic Kidney Disease and Peripheral Vascular Disease: A Light at the End of the Tunnel? Biomolecules 10 , doi: 10.3390/biom10010154 (2020). Wang, P., Xu, J., Wang, Y. & Cao, X. An interferon-independent lncRNA promotes viral replication by modulating cellular metabolism. Science (New York, N.Y.) 358 , 1051–1055, doi: 10.1126/science.aao0409 (2017). Giachelli, C. M. Vascular calcification mechanisms. J Am Soc Nephrol 15 , 2959–2964 (2004). Ta, N. N., Li, Y., Schuyler, C. A., Lopes-Virella, M. F. & Huang, Y. DPP-4 (CD26) inhibitor alogliptin inhibits TLR4-mediated ERK activation and ERK-dependent MMP-1 expression by U937 histiocytes. Atherosclerosis 213 , 429–435, doi: 10.1016/j.atherosclerosis.2010.08.064 (2010). Lv, J. et al. Long noncoding RNA H19-derived miR-675 aggravates restenosis by targeting PTEN. Biochem Biophys Res Commun 497 , 1154–1161, doi: 10.1016/j.bbrc.2017.01.011 (2018). Balderman, J. A. F. et al. Bone morphogenetic protein-2 decreases microRNA-30b and microRNA-30c to promote vascular smooth muscle cell calcification. Journal of the American Heart Association 1 , e003905, doi: 10.1161/JAHA.112.003905 (2012). Zhang, L. et al. H19 knockdown suppresses proliferation and induces apoptosis by regulating miR-148b/WNT/β-catenin in ox-LDL -stimulated vascular smooth muscle cells. J Biomed Sci 25 , 11, doi: 10.1186/s12929-018-0418-4 (2018). Mayer, O. [Matrix Gla protein as natural inhibitor of vascular calcification and potential treatment target]. Casopis lekaru ceskych 155 , 13–16 (2016). Touyz, R. M. et al. Vascular smooth muscle contraction in hypertension. Cardiovasc Res 114 , 529–539, doi: 10.1093/cvr/cvy023 (2018). Fragiadaki, M. & Mason, R. M. Epithelial-mesenchymal transition in renal fibrosis - evidence for and against. Int J Exp Pathol 92 , 143–150, doi: 10.1111/j.1365-2613.2011.00775.x (2011). Eddy, A. A. Overview of the cellular and molecular basis of kidney fibrosis. Kidney Int Suppl (2011) 4 , 2–8 (2014). Röhrborn, D., Wronkowitz, N. & Eckel, J. DPP4 in Diabetes. Frontiers in immunology 6 , 386, doi: 10.3389/fimmu.2015.00386 (2015). Fan, S.-H., Xiong, Q.-F., Wang, L., Zhang, L.-H. & Shi, Y.-W. Glucagon-like peptide 1 treatment reverses vascular remodelling by downregulating matrix metalloproteinase 1 expression through inhibition of the ERK1/2/NF-κB signalling pathway. Molecular and cellular endocrinology 518 , 111005, doi: 10.1016/j.mce.2020.111005 (2020). Cao, X., Yeo, G., Muotri, A. R., Kuwabara, T. & Gage, F. H. Noncoding RNAs in the mammalian central nervous system. Annual review of neuroscience 29 (2006). Mattick, J. S. & Makunin, I. V. Non-coding RNA. Human molecular genetics 15 Spec No 1 , R17-R29 (2006). Amaral, P. P. & Mattick, J. S. Noncoding RNA in development. Mammalian genome: official journal of the International Mammalian Genome Society 19 , 454–492, doi: 10.1007/s00335-008-9136-7 (2008). Costa, F. F. Non-coding RNAs, epigenetics and complexity. Gene 410 , doi: 10.1016/j.gene.2007.12.008 (2008). Ghildiyal, M. & Zamore, P. D. Small silencing RNAs: an expanding universe. Nature reviews. Genetics 10 , doi: 10.1038/nrg2504 (2009). Gu, X., Li, M., Jin, Y., Liu, D. & Wei, F. Identification and integrated analysis of differentially expressed lncRNAs and circRNAs reveal the potential ceRNA networks during PDLSC osteogenic differentiation. BMC Genet 18 , 100, doi: 10.1186/s12863-017-0569-4 (2017). Pardo, A. & Selman, M. MMP-1: the elder of the family. Int J Biochem Cell Biol 37 , 283–288 (2005). Hayami, T., Kapila, Y. L. & Kapila, S. MMP-1 (collagenase-1) and MMP-13 (collagenase-3) differentially regulate markers of osteoblastic differentiation in osteogenic cells. Matrix Biol 27 , 682–692, doi: 10.1016/j.matbio.2008.07.005 (2008). Lehrke, M. et al. MMP-1 serum levels predict coronary atherosclerosis in humans. Cardiovascular diabetology 8 , 50, doi: 10.1186/1475-2840-8-50 (2009). Kessenbrock, K., Wang, C.-Y. & Werb, Z. Matrix metalloproteinases in stem cell regulation and cancer. Matrix Biol 44–46 , 184–190, doi: 10.1016/j.matbio.2015.01.022 (2015). Ngai, D., Lino, M. & Bendeck, M. P. Cell-Matrix Interactions and Matricrine Signaling in the Pathogenesis of Vascular Calcification. Front Cardiovasc Med 5 , 174, doi: 10.3389/fcvm.2018.00174 (2018). Dong, C. et al. Anagliptin stimulates osteoblastic cell differentiation and mineralization. Biomed Pharmacother 129 , 109796, doi: 10.1016/j.biopha.2019.109796 (2020). Röhrborn, D., Eckel, J. & Sell, H. Shedding of dipeptidyl peptidase 4 is mediated by metalloproteases and up-regulated by hypoxia in human adipocytes and smooth muscle cells. FEBS Lett 588 , 3870–3877, doi: 10.1016/j.febslet.2014.08.029 (2014). Tables Table 1. The characteristics of the lincRNAs induced by DPP4. top 10 downregulated LincRNAs Seqname Chrom GeneSymbol Fold Change P-value RNA length Nearby Gene Symbol ENST00000540293 chr11 AP000619.5 5.4691462 0.018646883 1000 MMP1 uc001phf.3 chr11 AB231724 5.3684649 0.000377659 157 MMP1 uc002ywn.1 chr21 AL109792 4.0931103 0.002062516 2900 KCNJ6 T240875 Chr3 G055454 3.9343595 0.003993263 8371 LYZL4 T240875 Chr3 G055454 3.9343595 0.003993263 8371 VIPR1 T046285 Chr10 G010780 3.4726155 0.012499379 1603 CEP55 ENST00000412427 Chr1 RP11-380J14.1 3.3043558 0.002605688 4676 ATP13A2 ENST00000452326 Chr22 AC000067.1 3.2458684 0.012123994 287 C22orf29 ENST00000452326 Chr22 AC000067.1 3.2458684 0.012123994 287 CDC45 uc002xuq.1 Chr20 AK055386 2.9207398 0.008848319 3404 KCNB1 top 8 upregulated LincRNAs Seqname Chrom Gene Symbol Fold Change P-value RNA length Nearby Gene Symbol ENST00000450216 Chr22 CTA-150C2.13 4.1538977 0.033626171 678 APOBEC3B ENST00000450216 Chr22 CTA-150C2.13 4.1538977 0.033626171 678 APOBEC3A ENST00000607314 Chr8 RP11-1002K11.1 3.3245744 0.000171600 1835 NRG1 NR_003133 Chr1 GBP1P1 3.035663 0.017197101 1234 LRRC8B ENST00000520544 Chr8 RP11-760H22.2 2.2794756 0.006961939 490 COL14A1 ENST00000562082 Chr10 RP11-119F7.5 2.0858913 0.009993759 2295 STOX1 T365864 Chr9 G086505 2.0357136 0.033876979 1819 TRAF1 ENST00000608605 Chr3 RP11-804H8.6 2.0171033 0.006191725 2060 CISH Table 2 .The specific primers of LncRNA ENST00000540293,MMP-1 andβ-actin were designed. Gene name Primers sequence LncRNA ENST00000540293 Forward 5'-CTGGACCAGGTATCGGAGGAGATG-3' Reverse 5'-GGTAGCCACTCAGCAGTTGATCC-3' MMP-1 Forward 5'-AGATTCTACATGCGCACAAATC-3' Reverse 5'-CCTTTGAAAACCGGACTTCAT-3' β-actin Forward 5'-CCTGGCACCCAGCACAAT-3' Reverse 5'-GCCGATCCACACGGAGTA-3' Table 3. The characteristics of the mRNAs induced by DPP4 top 10 downregulated mRNAs Seqname Chrom RNA length Fold Change P-value Gene Symbol type NM_144634 chr3 916 7.7044098 0.001159771 LYZL4 protein_coding NM_198404 chr13 2133 5.2321373 0.003622246 KCTD4 protein_coding NM_003280 chr3 705 5.1611148 0.002562575 TNNC1 protein_coding NM_002240 chr21 2790 4.9792082 0.008725652 KCNJ6 protein_coding NM_005582 chr5 2725 4.8654311 0.003843377 CD180 protein_coding NM_003783 chr1 3548 4.7786871 0.001342022 B3GALT2 protein_coding NM_145060 chr18 2893 4.6475968 0.011197155 SKA1 protein_coding NM_021158 chr20 2554 4.5764374 0.011266314 TRIB3 protein_coding NM_002421 chr11 2081 4.2991079 0.028861407 MMP1 protein_coding NM_002849 chr12 3516 4.2792469 0.004702521 PTPRR protein_coding top 10 upregulated mRNAs Seqname Chrom RNA length Fold Change P-value Gene Symbol type NM_000064 chr19 5148 27.6095442 0.002442785 C3 protein_coding NM_001080400 chr19 6353 11.528119 0.001785257 PLIN4 protein_coding NM_000668 chr4 2707 11.3082146 0.015208381 ADH1B protein_coding NM_000014 chr12 4678 11.2869199 0.000195162 A2M protein_coding NM_000669 chr4 1769 11.1388959 0.00879313 ADH1C protein_coding NM_013244 chr12 1934 9.4263368 0.011624943 MGAT4C protein_coding NM_000587 chr5 4034 7.0028976 0.023068264 C7 protein_coding NM_004000 chr1 1486 6.8544351 0.020855768 CHI3L2 protein_coding NM_000041 chr19 1234 6.70272 0.000195164 APOE protein_coding NM_005525 chr1 1477 6.6150899 0.017043214 HSD11B1 protein_coding Additional Declarations No competing interests reported. Supplementary Files Supplementalfiguresandlegends.pdf SupplementaryDatasetFile1DifferentiallyExpressedLncRNAs.xls SupplementaryDatasetFile2DifferentiallyExpressedmRNAs.xls SupplementaryDatasetFile3LincRNAsassociatedcodinggenedatatable.xls Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-1656423","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":110462187,"identity":"fd0a0f03-f883-4239-b42d-b50b935c9279","order_by":0,"name":"Tongjie Xu","email":"","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Tongjie","middleName":"","lastName":"Xu","suffix":""},{"id":110462188,"identity":"28bfd8b7-7012-4d75-a6e5-7bb0cccb53d9","order_by":1,"name":"Hao Chen","email":"","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hao","middleName":"","lastName":"Chen","suffix":""},{"id":110462189,"identity":"0bd32c6e-2014-4bb2-94d1-532931ba7eba","order_by":2,"name":"Xiongfei Xu","email":"","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiongfei","middleName":"","lastName":"Xu","suffix":""},{"id":110462190,"identity":"0a75d399-1894-47ca-b06b-49cd3c25bfb4","order_by":3,"name":"Xiaolei Sun","email":"","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Xiaolei","middleName":"","lastName":"Sun","suffix":""},{"id":110462191,"identity":"a946da2d-af00-4795-8179-e4c89154ad51","order_by":4,"name":"Huqiang He","email":"","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Huqiang","middleName":"","lastName":"He","suffix":""},{"id":110462192,"identity":"8fa19a54-9621-4e4b-bbe1-30dbe59c7cc5","order_by":5,"name":"Hong Zeng","email":"","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Hong","middleName":"","lastName":"Zeng","suffix":""},{"id":110462193,"identity":"627c3217-7c01-4128-9490-5477ca1c255b","order_by":6,"name":"Sen Shi","email":"","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":false,"submittingAuthor":false,"prefix":"","firstName":"Sen","middleName":"","lastName":"Shi","suffix":""},{"id":110462194,"identity":"a48ab0f4-76b5-4d48-a670-6e987a9cf3e6","order_by":7,"name":"Yong Liu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAtklEQVRIiWNgGAWjYBACPhDB+I9Njo29/QBxWtggJJ8xH8+ZBJK0yCXOk3AwIFKLRPKxh194zNLbJBgSGH5UbCNGS1q6sYxEWm6bdOMBxp4zt4nRkmMmLWFwLLdN5kACM2Mb0VoS/qezSSQYEK9F8sMBtgQStPA8S5NmbGAzbAMG8kGi/MLPnnxM8mcDm7x8e/vBBz8qiNDCIJDAwMwDZR8gQj3ImgMMjD+IUzoKRsEoGAUjFQAAcoU0MmDINK8AAAAASUVORK5CYII=","orcid":"","institution":"The Affiliated Hospital of Southwest Medical University","correspondingAuthor":true,"submittingAuthor":false,"prefix":"","firstName":"Yong","middleName":"","lastName":"Liu","suffix":""}],"badges":[],"createdAt":"2022-05-14 14:44:07","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-1656423/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-1656423/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":22242903,"identity":"b3e8f0e9-5cea-4483-9c13-292abdea8164","added_by":"auto","created_at":"2022-06-03 22:21:55","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2531652,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(A)\u003c/strong\u003e The expression of calcification-related protein OPG, OPN, Runx2 and BMP-2 with intervention factor of DPP4(*P\u0026lt;0.05, **P\u0026lt;0.01, ***P\u0026lt;0.001). \u003cstrong\u003e(B)\u003c/strong\u003e and \u003cstrong\u003e(D)\u003c/strong\u003e \u003cstrong\u003e.\u003c/strong\u003eThe calcification of each group was examined by Alizarin red staining.Orange-red calcified nodules in the DPP4 group were significantly more than the control Group. \u003cstrong\u003e(C) \u003c/strong\u003eCalcium content was measured by Ca determination kit.The intracellular calcium content in DPP4 group increased significantly (***P\u0026lt;0.001).The data are presented as meanent in each group (n=3) of the two independent experiments.\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/b1ca69a6bebeadf27a7b3b20.png"},{"id":22242910,"identity":"6f228646-4680-480f-8fe6-9f76b7e827de","added_by":"auto","created_at":"2022-06-03 22:21:56","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":486658,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(A) \u003c/strong\u003eExpression profiles of lncRNAs and mRNAs in HASMCs after DPP4 intervention. Hierarchical clustered heat maps of differentially expressed lncRNAs and mRNAs. \u003cstrong\u003e(B) \u003c/strong\u003eVolcano plots were generated to visualize the differentially expressed lncRNAs and mRNAs. The X-axis indicates the log2 fold-change and the Y-axis represents the log10 P-values. The red and green points indicate the upregulated and downregulated lncRNAs and mRNAs with statistica significance, respectively.The data are presented as mean±SE in each group (n=3) of the three independent experiments.\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/d7af58d50853b318767148b8.png"},{"id":22242911,"identity":"51e50db1-8db6-4629-aed6-841a0a2ed0f3","added_by":"auto","created_at":"2022-06-03 22:21:57","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":505609,"visible":true,"origin":"","legend":"\u003cp\u003eThe level change of several calcification proteins after lncRNA ENST00000540293 knockdown in HASMCs. Among them, the levels of BMP-2\u003cstrong\u003e(A)\u003c/strong\u003e,OPG\u003cstrong\u003e(B)\u003c/strong\u003e,OPN\u003cstrong\u003e(C)\u003c/strong\u003e and RUNX2\u003cstrong\u003e(D)\u003c/strong\u003e statistically significantly increased after lncRNA ENST00000540293 was knocked down(**p\u0026lt;0.01,***p\u0026lt;0.001).The data are presented as meanent in each group (n=3) of four independent experiments.\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/ca1f2b62a649551d02151cb7.png"},{"id":22242912,"identity":"5ef8848e-d0ac-4d94-baea-d23f64a57456","added_by":"auto","created_at":"2022-06-03 22:21:57","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":5593781,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003e(A) \u003c/strong\u003eand\u003cstrong\u003e (B) \u003c/strong\u003eThe calcification of each group was examined by Alizarin red staining.Orange-red calcified nodules in the siRNA group were significantly more than other Groups.The data are presented as mean±SE in each group of the three independent experiments. \u003cstrong\u003e(C) and\u003c/strong\u003e \u003cstrong\u003e(D) T\u003c/strong\u003ehe activity of ALP and intracellular calcium content in siRNA group increased significantly (***P\u0026lt;0.001).The data are presented as meanent in each group (n=3) of three independent experiments.\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/708eb6d8180891f518d4e679.png"},{"id":22242907,"identity":"e7c121d7-8d70-49e7-9e06-d8835327daf6","added_by":"auto","created_at":"2022-06-03 22:21:56","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1929567,"visible":true,"origin":"","legend":"\u003cp\u003eThe expression of α-SMA protein in HASMCs was detected by Western blot. \u003cstrong\u003e(A)\u003c/strong\u003e shows the relative expression of α-SMA protein, and \u003cstrong\u003e(B) \u003c/strong\u003eshows the F-actin staining ratio of actin induced by cyclopeptide. In siRNA group, the expression of α-SMA protein decreased significantly (**P\u0026lt;0.01), while the expression of F-actin decreased and aggregation decreased. \u003c/p\u003e","description":"","filename":"Figure5.png","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/808859b4889bc49fb02bc582.png"},{"id":22242904,"identity":"773786ef-82b5-470b-b455-81bc642045c3","added_by":"auto","created_at":"2022-06-03 22:21:56","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":379856,"visible":true,"origin":"","legend":"\u003cp\u003eThe expression of MMP-1\u003cstrong\u003e(A)\u003c/strong\u003e and ERK1/2\u003cstrong\u003e(B)\u003c/strong\u003e in HASMCs was detected by Western blot. Compared with the other groups, MMP-1 protein and ERK1/2 phosphorylate in siRNA group increased significantly, and the difference was statistically significant.( **P＜0.01,***P＜0.001).\u003c/p\u003e","description":"","filename":"Figure6.png","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/59c757c2ed777b7bbbf30e12.png"},{"id":24176888,"identity":"0363a33c-4589-4ac8-9510-329bf7af5eb4","added_by":"auto","created_at":"2022-07-22 07:14:32","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2859712,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/78f9aa82-a709-47f2-8530-79c967535134.pdf"},{"id":22242905,"identity":"9163746f-8774-46c1-94c9-a0912dbc6d04","added_by":"auto","created_at":"2022-06-03 22:21:56","extension":"pdf","order_by":12,"title":"","display":"","copyAsset":false,"role":"supplement","size":1034789,"visible":true,"origin":"","legend":"","description":"","filename":"Supplementalfiguresandlegends.pdf","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/d333eaede6a218180ec81521.pdf"},{"id":22243279,"identity":"9f4a0b8c-1e13-4522-9543-da881c90d695","added_by":"auto","created_at":"2022-06-03 22:26:57","extension":"xls","order_by":13,"title":"","display":"","copyAsset":false,"role":"supplement","size":486400,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryDatasetFile1DifferentiallyExpressedLncRNAs.xls","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/3f102cb196f39a5c1b2fcaae.xls"},{"id":22242909,"identity":"bd2fcbd0-3675-46db-a3b6-95af51cf7540","added_by":"auto","created_at":"2022-06-03 22:21:56","extension":"xls","order_by":14,"title":"","display":"","copyAsset":false,"role":"supplement","size":605696,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryDatasetFile2DifferentiallyExpressedmRNAs.xls","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/ad9e72cdfcd1533f6e60b67b.xls"},{"id":22243278,"identity":"563d5f54-f661-4e4c-a037-d8a21260c869","added_by":"auto","created_at":"2022-06-03 22:26:56","extension":"xls","order_by":15,"title":"","display":"","copyAsset":false,"role":"supplement","size":39424,"visible":true,"origin":"","legend":"","description":"","filename":"SupplementaryDatasetFile3LincRNAsassociatedcodinggenedatatable.xls","url":"https://assets-eu.researchsquare.com/files/rs-1656423/v1/b90b71c836807865bc8c189e.xls"}],"financialInterests":"No competing interests reported.","formattedTitle":"Expression profile of lncRNA induced by DPP4 in human aortic smooth muscle cells and calcification gene prediction","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eVascular calcification is a common pathological phenomenon in coronary atherosclerotic heart disease, chronic kidney disease, diabetes and other diseases\u003csup\u003e1\u003c/sup\u003e,\u0026nbsp;Its pathological basis is an expression of osteoblast-specific genes in smooth muscle cells\u003csup\u003e2\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e Previous studies have shown that vascular calcification is a very complex pathological process, which is regulated by many factors.\u0026nbsp;Recent studies have found that vascular calcification is an adjustable and active biological process, which is similar to bone formation and involves the expression of some bone formation related proteins, such as alkaline phosphatase (ALP), osteocalcin (OCN), Runx2 and BMP-2\u003csup\u003e3\u003c/sup\u003e.\u0026nbsp;Vascular calcification is common in patients with diabetes, and the important pathological change of vascular complications is intima-media calcification. Medial calcification and cardiovascular disease caused by diabetes are one of the main causes of death. Vascular calcification is closely related to the high incidence and mortality of cardiovascular disease. Therefore, it is of great significance to study the treatment and intervention of diabetic vascular calcification.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eDipeptidyl peptidase 4 (DPP4) is a highly glycosylated type II transmembrane protein composed of 766 amino acids, which belongs to the family of serine proteases. One of its catalytic substrates is incretin. The treatment of type 2 diabetes with DPP4 inhibitors (Letins) establishes the role of incretin in blood glucose control.\u0026nbsp;Low molecular weight DPP4 inhibitors exert their hypoglycemic effect by promoting glucose-dependent insulinotropic polypeptide (GIP) and inhibiting the degradation of glucagon-like peptide 1 (GLP-1). DPP4 is widely expressed in a variety of cells, such as endothelial cells and immune cells. DPP4 also plays an important role in the regulation of cardiovascular function\u003csup\u003e4\u003c/sup\u003e.\u0026nbsp;Meanwhile, DPP4 not only plays an important role in reducing blood sugar, but also has unique functions in other aspects\u003csup\u003e5\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e Nina Wronkowitz et al demonstrated that soluble DPP4 induces inflammation and proliferation of human smooth muscle cells via protease-activated receptor 2\u003csup\u003e6\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e Yu et al demonstrated that soluble DPP4 can induce oxidative stress in human umbilical vein endothelial cells through mannose 6-phosphate/insulin-like growth factor receptor II\u003csup\u003e7\u003c/sup\u003e.\u0026nbsp;A.M.Lambeir et al found that the expression of DPP4 in visceral tissues of obese and diabetic patients was higher than normal people\u003csup\u003e8\u003c/sup\u003e. Studies have shown that DPP4 and its inhibitors are closely related to vascular calcification\u003csup\u003e9\u003c/sup\u003e. DPP4 may lead to vascular calcification\u003csup\u003e10\u003c/sup\u003e. While,DPP4 inhibitors may lowered the levels of fasting serum glucose, triglyceride (TG),TNF-\u0026alpha; and decreased the calcium deposits\u003csup\u003e11\u003c/sup\u003e, and also attenuated vascular calcification and osteogenic trans-differentiation in vascular smooth muscle cells via multiple steps, such as: downregulation of PiT-1 expression and suppression of reactive oxygen species generation, phospho-PI3K/AKT, and the Wnt signaling pathway\u003csup\u003e12\u003c/sup\u003e.The above data suggest that diabetes and its concomitant diseases may be closely related to the regulation of DPP4.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eLong non-coding RNA (lncRNA) is a functional RNA molecule with a length of more than 200bp in non-coding RNA\u003csup\u003e13\u003c/sup\u003e.\u0026nbsp;LncRNA is located in the cytoplasm or nucleus. They regulate gene expression through epigenetic, transcriptional and post-transcriptional regulation in the form of RNA, but lack the ability of protein coding. It has been found that lncRNA plays an important role in regulating cell growth, development, distribution and targeted differentiation\u003csup\u003e14\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e Intergenic lncRNA(lincRNA) and antisense lncRNA are one of the hotspots in the current research.\u0026nbsp;LincRNA ENST00000540293 is an intergenic noncoding RNA located on chromosome 11 and contains 1000 bases, and may associated with human periodontal ligament cells development\u003csup\u003e15\u003c/sup\u003e. Our RNA microarray results show that the lncRNA ENST00000540293 nearby gene is NM-002421, and the protein encoded by NM-002421 is matrix metalloproteinase 1 (MMP-1)\u003cstrong\u003e\u0026nbsp;(\u003c/strong\u003e\u003cstrong\u003eTable 1\u003c/strong\u003e\u003cstrong\u003e)\u003c/strong\u003e\u003cstrong\u003e\u003csup\u003e16\u003c/sup\u003e\u003c/strong\u003e.\u0026nbsp;Previous studies have shown that matrix metalloproteinases are closely related to the occurrence and development of vascular calcification\u003csup\u003e17,18\u003c/sup\u003e.\u0026nbsp;Our preliminary experiments in vitro experiments showed that DPP4 can promote the calcification of HASMCs by activating the ERK/NF-kB pathway, and the DPP4 inhibitors (sitagliptin) can inhibit ERK1/2 and NF-kB signaling pathway so that inhibits aortic vascular calcification in type 2 diabetic mice also confirmed in vivo. Although DPP4 is closely related to cardiovascular disease, the exact mechanism of DPP4 promoting HASMCs calcification is not completely clear\u003csup\u003e11\u003c/sup\u003e. Therefore, in this study, HASMCs were cultured with DPP4 to establish calcification model, so as to study the mechanism of DPP4 promoting osteogenic differentiation of vascular smooth muscle cells.\u0026nbsp;\u003c/p\u003e"},{"header":"2. Materials And Methods","content":"\u003cp\u003e\u003cstrong\u003e2.1 The culture of HASMCs\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe primary human aortic vascular smooth muscle cell (HASMCs,no.6110) and culture medium (no.1101)were obtained from Science Cell Research-Laboratory (USA). SMCM medium was composed of 500ml basic medium, 10ml fetal bovine serum (FBS, no.0010), 5ml smooth muscle cell growth factor (SMCGS, no.1152) and 5ml penicillin / streptomycin (Pmax S, no.0503).The 3rd to 8th generation cells were selected for the experiment and cultured in an incubator at 37℃with 5% humidified CO2.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.2 Determination of intracellular calcium concentration\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe cell intracellular calcium concentration was determined using a spectrophotometer at 570 nm. Transferring 800ul of the buffer to another 1 ml cuvette and adding 100 ul of the reaction solution by pipette. They were evenly mixed and incubated at room temperature for 5 minutes. Taking 100 \u0026mu;l mixture liquid in 96-well plates with 3-6 multiple holes in each group. The standard curve was plotted with the optical density (OD) on the y axis and the standard calcium concentration (mmol/l) on the x axis and get the calcium concentration.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.3 Western blotting\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe levels of the all target proteins was measured by the fluorescence imaging system Image Studio, and the expression level of protein was quantified by measuring the greyscale of bands and normalized to \u0026beta;-actin. Briefly,the concentration of protein was detected using a BCA kit (Beyotime Biotechnology, Shanghai,China). Then, Approximately 20\u0026mu;g\u0026nbsp;of protein was loaded onto a 10% SDS-PAGE gel and blotted onto PVDF membrane.After blocking with 5% BSA, the membranes were incubated with primary antibodies at\u0026nbsp;4\u0026deg;C\u0026nbsp;overnight. The membranes were washed thrice by TBST and incubated with secondary antibodies for 1 h at room temperature.Such as Osteoprotegerin (OPG), Bone Morphogenetic Protein 2 (BMP-2), Osteopontin (OPN), Runt Related Transcription Factor 2 (RUNX2), \u0026alpha; Smooth Muscle actin (\u0026alpha;-SMA), Matrix Metalloproteinase 1 (MMP-1) were detected by western blotting with their respective antibodies(anti-OPG,OPN,BMP-2 ,MMP-1 all from Abcam; anti-\u0026alpha;-SMA from R\u0026amp;D Systems;anti-RUNX2 from Cell Signaling Technology).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.4 Alizarin red staining for Calcification Detection\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eCell calcification nodules were detected by Alizarin red kit（Sigma USA)）.Control group and experimental group cells were inoculated on 12-well plates. Cleaning solution was added to each well to clean the growing cell surface, fixed solution was added and incubated at room temperature for 15 minutes, then staining solution was added and incubated at room temperature for 5 minutes or until orange-red was visible. The staining solution was removed and dried in air at room temperature for about 10 minutes. Then transparent solution was added and immediately observed under light microscope: the positive cells of calcium deposition were orange-red.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.5 F-actin staining\u0026nbsp; \u0026nbsp;\u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp; \u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe HASMCs (2\u0026times;10\u003csup\u003e5\u003c/sup\u003e cells/well) were inoculated on a 6-well plate overnight and then treated with the tested compound for 72 hours. Cells were washed with PBS and fixed with 4% Formaldehyde, Permeabilized in 0.2% Triton X-100 at room temperature for 5 minutes, then stained with DAPI (Beyotime,USA) and Rhodamine Phalloidin (1:200,Sigma,MO,USA) to mark the cyteblast and cytoskeleton respectively. Pictures were acquired by Photometrics CoolSNAPHQ2 CCD camera on the Olympus IX71-Applied Precision Delta Vision restoration microscope (Applied Precision, USA) and deconvolved using Delta Vision algorithms (Applied Precision, USA).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.6 Detection of intracellular ALP activity\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eFor ALP activity measurement,the calcified HASMCs were solubilized with RIPA lysis buffer (Beyotime Biotechnology, Shanghai,China). After centrifugation, the supernatants were examined with the ALP activity kit (Beyotime Biotechnology,Shanghai,China) .The optical density (OD) value of each group was determined by 450nm wavelength. The standard curve was made and the corresponding ALP activity of each group was calculated.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.7\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;RNA extraction and real-time quantitative polymerase chain reaction.\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eTotal RNA was extracted using TRI zol reagent (Invitrogen;Thermo Fisher Scientific, Inc).\u0026nbsp;RNA quantity and quality were measured by Nano Drop ND-1000. RNA integrity was assessed by standard denaturing agarose gel electrophoresis. Based on the predicted consequences, we selected the target linRNA with the most expression change between the control group and DPP4 intervened group and designed PCR primers (\u003cstrong\u003eTable 2\u003c/strong\u003e). RT‑qPCR was performed using SYBR Green Real‑Time PCR Master mix (Invitrogen;Thermo Fisher Scientific, Inc),and was run at 95˚C for 5 min, followed by 40 cycles at 95˚Cfor 10 sec, 65˚C for 20 sec and 72˚C for 30 sec. The characteristics of the linRNAs and mRNAs are presented in \u003cstrong\u003eTable 1 and Table 3\u003c/strong\u003e. Data were normalized to \u0026beta;-actin and the relative level of gene expression was calculated using the 2\u003csup\u003e-\u0026Delta;\u0026Delta;Ct\u0026nbsp;\u003c/sup\u003emethod.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.8 Microarray\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eArraystar Human LncRNA Microarray V4.0 is designed for the global profiling of human lncRNAs and protein-coding transcripts, which is updated from the previous Microarray V3.0. About 40,173 lncRNAs and 20,730 coding transcripts can be detected by our third-generation lncRNA microarray.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.9 RNA labeling and array hybridization\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSample labeling and array hybridization were performed according to the Agilent One-Color Microarray-Based Gene Expression Analysis protocol (Agilent Technology) with minor modifications. Briefly, mRNA was purified from total RNA after removal of rRNA (mRNA-ONLY Eukaryotic mRNA Isolation Kit, Epicentre). Then, each sample was amplified and transcribed into fluorescent cRNA along the entire length of the transcripts without 3\u0026apos; bias utilizing a random priming method (Arraystar Flash RNA Labeling Kit, Arraystar). The labeled cRNAs were purified by RNeasy Mini Kit (Qiagen). The concentration and specific activity of the labeled cRNAs (Pmol Cy3/\u0026mu;g cRNA) were measured by NanoDrop ND-1000. 1 \u0026mu;g of each labeled cRNA was fragmented by adding 5 \u0026mu;l 10 \u0026times; Blocking Agent and 1 \u0026mu;l of 25 \u0026times; Fragmentation Buffer, then heated the mixture at 60\u0026deg;C for 30 min, finally 25 \u0026mu;l 2 \u0026times; GE Hybridization buffer was added to dilute the labeled cRNA. 50 \u0026mu;l of hybridization solution was dispensed into the gasket slide and assembled to the LncRNA expression microarray slide. The slides were incubated for 17 hours at 65\u0026deg;C in an Agilent Hybridization Oven. The hybridized arrays were washed, fixed and scanned with using the Agilent DNA Microarray Scanner (part number G2505C).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.10 LncRNA transient transfection.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eVSMCs were interference with 50 nm lncRNA ENST00000540293 Smart Silencer (siRNA ) and negative control (NC-siRNA) using Smart Silencer NC at a density of 4 x10\u003csup\u003e5\u003c/sup\u003e cells \u0026nbsp; for 48 hours, according to the manufacturer\u0026apos;s protocol\u003csup\u003e19\u003c/sup\u003e. The inhibitory efficiency was tested by real-time qPCR, and the effective siRNA was picked for the further experiments.\u0026nbsp;The lncRNA ENST00000540293 siRNA and NC-siRNA were synthesized by RiboBioCo (Guangzhou, China).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eStatistical analysis.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData are presented as the mean \u0026plusmn; standard deviation (n=3) and were analyzed using a Student\u0026apos;s t-test. All statistical analysis was performed with SPSS 20.0 (IBM Corp, Armonk, NY, USA). P value \u0026lt; 0.05 was accepted as significant.\u003c/p\u003e"},{"header":"3. Results","content":"\u003cp\u003e\u003cstrong\u003e1\u003c/strong\u003e\u003cstrong\u003e.Effect of DPP4 on HASMCs calcification protein\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe 5rd to 7th generation HASMCs were cultured with 200ng/ml DPP4 for 7 days. The total cell protein was extracted and the expression of OPG, OPN, BMP-2 and RUNX2 was detected by western blotting. The expression of OPG, OPN, BMP-2 and RUNX2 protein in DPP4 group was significantly higher than control group (\u003cstrong\u003eFig.1A\u003c/strong\u003e).\u0026nbsp;The results confirm that DPP4 can induce HASMCs calcification.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2\u003c/strong\u003e\u003cstrong\u003e.Calcification of HASMCs induced by DPP4\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter cultured with 200ng/ml DPP4 for 14 days, the cells in the control group showed typical multi-layer smooth muscle cells, and the alizarin red staining was negative, while the orange red calcified nodules in the DPP4 group were significantly more than those in the control group.The alizarin red staining was strongly positive (\u003cstrong\u003eFig.1Ban Fig.1D\u003c/strong\u003e),and calcium content was measured(\u003cstrong\u003eFig.1C)\u003c/strong\u003e, which further confirmed that DPP4 could induce HASMCs calcification.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.Expression profile of lncRNA and mRNA in HASMCs after DPP4 intervention\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAfter adding 200ng/ml DPP4 to HASMCs, the total RNA was extracted to detect its quality, concentration and integrity.Differentially expressed lncRNAs and mRNAs with statistical significance between the two groups were identified through P-value/FDR filtering. Differentially expressed lncRNAs and mRNAs between the two samples were identified through Fold Change filtering(\u0026nbsp;\u003cstrong\u003eFig\u003c/strong\u003e\u003cstrong\u003e.2A and\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eFig\u003c/strong\u003e\u003cstrong\u003e.2B\u003c/strong\u003e).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e4.DPP4 inhibits the expression of lncRNA ENST00000540293 and MMP-1 mRNA in HASMCs\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe differentially expressed lncRNAs or mRNAs between the two groups were screened by P-value/FDR, and further screened by Fold Change. The results suggest that DPP4 can inhibit the expression of lncRNA ENST00000540293 and MMP-1 mRNA (\u003cstrong\u003eTable 1 and Table3 ).\u003c/strong\u003eTo confirm the reliability and validity of the microarray data. LncRNAs and\u0026nbsp;mRNAs\u0026nbsp;chip screening results were verified by RT-qPCR.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e5.Effect of inhibiting the expression of HASMCs lncRNA ENST00000540293 on calcification proteins.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp; In order to clarify the relationship between inhibition of lncRNA ENST00000540293 expression and calcification-related protein expression, HASMCs was divided into control group, transfection control group(include Transfection Reagent only), siRNA control group and siRNA group and cultured for 48 hours. The expression of calcification-related proteins BMP-2, OPG, OPN and RUNX2 was detected by Western blot. The results showed that the expression of calcitonin in siRNA group was significantly higher than the other groups (\u003cstrong\u003eFig.3\u003c/strong\u003e). It was proved that inhibiting the expression of lncRNA ENST00000540293 could promote the expression of HASMCs calcitonin.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e6. Morphological analysis and calcium content determination of calcification of HASMCs after inhibition of lncRNA ENST00000540293 expression.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eIn order to further analyze the cell morphology, intracellular calcium content and the activity of alkaline phosphatase (ALP) after inhibiting the expression of lncRNA ENST00000540293 in HASMCs.\u0026nbsp;HASMCs\u0026nbsp;were cultured for 72 hours and stained with alizarin red. The intracellular calcium content, activity of ALP of each group was determined. The results showed : the area of calcified nodules and orange calcium nodules increased significantly in SiRNA group(\u003cstrong\u003eFig.4A and Fig.4B\u003c/strong\u003e),the activity of ALP and intracellular calcium content in SiRNA group increased significantly (***P\u0026lt;0.001)(\u003cstrong\u003eFig. 4C and Fig.4D\u003c/strong\u003e). Previous studies have proved that there is a positive correlation between intracellular ALP activity, intracellular calcium content and\u0026nbsp;HASMCs\u0026nbsp;calcification. The results showed that inhibition of epidermis of lncRNA ENST00000540293 could promote the calcification of\u0026nbsp;HASMCs.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e7.Inhibition of lncRNA ENST 00000540293 expression promotes phenotypic transformation of HASMCs.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe mechanism of differentiation of HASMCs into osteoblast-like cells is similar to that of bone formation\u003csup\u003e2\u003c/sup\u003e\u003csup\u003e0\u003c/sup\u003e.In order to further verify that inhibiting the expression of lncRNA ENST00000540293 can promote the differentiation of VSMCs into osteoblasts, we detected the expression of \u0026alpha;-SMA by Western blot(\u003cstrong\u003eFig.5A\u003c/strong\u003e). The expression of \u0026alpha;-SMA protein in siRNA group was significantly lower than other groups (**P \u0026lt;0.01). At the same time, the results of F-actin immunofluorescence staining were observed by laser confocal microscope(\u003cstrong\u003eFig.5B\u003c/strong\u003e). F-actin (red) fluorescence intensity, myofilament expression and aggregation in siRNA group were significantly decreased, further indicate that HASMCs had a tendency to transform into osteoblasts. In short, it can be speculated that after lncRNA ENST00000540293 silencing, the phenotype of HASMCs changes from contractile type to synthetic secretory type, and finally into osteoblast-like cells.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e8. Silencing lncRNA ENST00000540293 promoted the expression of MMP-1 and\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eERK\u003csub\u003e1/2\u003c/sub\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003ein HASMCs.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eStudies have shown that DPP4 regulate the expression of MMP-1 protein through ERK1/2 signal pathway\u003csup\u003e2\u003c/sup\u003e\u003csup\u003e1\u003c/sup\u003e.And lncRNA may play a role by regulating coding genes near transcription or other signal way\u003csup\u003e2\u003c/sup\u003e\u003csup\u003e2\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e Our previous gene chip microarray subarray analysis showed that there were many differentially expressed lncRNAs near mRNAs related to tissue development, in which (Nearby Gene), a gene attached to lncRNA ENST00000540293, was a protein encoded by NM-002421, which was matrix metalloproteinase-1 (\u003cstrong\u003eTable 1\u003c/strong\u003e). It was found that after inhibiting the expression of lncRNA ENST00000540293, MMP-1 protein and ERK\u003csub\u003e1/2\u0026nbsp;\u003c/sub\u003ephosphorylate in siRNA group was significantly higher than the other groups detected by Western blot(\u003cstrong\u003eFig.6A and Fig.6B\u003c/strong\u003e),and the difference was statistically significant (**p\u0026lt;0.01,***p\u0026lt;0.001).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eVascular calcification is a pathological process in which HASMCs change from contractile type to synthetic secretory type and finally to osteoblast-like phenotype under various pathological actions, which mediates the abnormal deposition of calcium in the vascular wall. Vascular calcification exists widely in diabetes, atherosclerosis, hypertension, chronic renal failure and other diseases, and the degree of calcification is positively correlated with the severity of the disease\u003csup\u003e2\u003c/sup\u003e\u003csup\u003e3\u003c/sup\u003e\u003csup\u003e,2\u003c/sup\u003e\u003csup\u003e4\u003c/sup\u003e.\u0026nbsp;Previous studies have shown that the occurrence and development of vascular calcification is closely related to cell degeneration and necrosis, and it is a passive and degenerative vascular terminal lesion process, while recent studies have found that vascular calcification is a complex biological process that is controllable, active, preventable and reversible similar to bone formation\u003csup\u003e2\u003c/sup\u003e\u003csup\u003e5\u003c/sup\u003e. Changes in extracellular conditions and phenotypic transformation of vascular smooth muscle cells play a key role\u003csup\u003e2\u003c/sup\u003e\u003csup\u003e6\u003c/sup\u003e.\u0026nbsp;At the same time, vascular calcification is similar to intramembranous osteogenesis, which usually leads to an increase in vascular hardness, a decrease in vascular compliance, and an increase in the incidence and mortality of cardiovascular events, more common in diabetes, chronic kidney disease\u003csup\u003e2\u003c/sup\u003e\u003csup\u003e7\u003c/sup\u003e\u003csup\u003e,2\u003c/sup\u003e\u003csup\u003e8\u003c/sup\u003e．DPP4 also known as T cell surface antigen CD26, containing 110kDa glycoprotein, is a serine protease widely expressed on the surface of all kinds of cells. As a new type of fat factor, it can decompose and metabolize glucagon-like peptide-1 (GLP-1) to regulate blood sugar, and also plays an important role in the pathogenesis of diabetes and cardiovascular disease\u003csup\u003e29\u003c/sup\u003e\u003csup\u003e,3\u003c/sup\u003e\u003csup\u003e0\u003c/sup\u003e.\u0026nbsp;At the same time, it has been found that DPP4 as an agonist of protease activated receptor 2 (PAR2), can directly activate the cascade of MAPK and NF-kB signals involved in PAR2, and induce the proliferation and inflammation of\u0026nbsp;HASMCs\u003csup\u003e6\u003c/sup\u003e.\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp;Long non-coding RNA is a functional RNA transcribed by RNA polymerase II. It is located in the nucleus or cytoplasm. Due to the lack of the ability to encode proteins, it was once considered to have no biological function.\u0026nbsp;At present, it is believed that it plays a similar role in the regulation of gene expression. Such as chromosome silencing, chromatin modification, transcriptional activation, transcriptional interference, nuclear transport and other important regulatory processes\u003csup\u003e3\u003c/sup\u003e\u003csup\u003e1\u003c/sup\u003e\u003csup\u003e-3\u003c/sup\u003e\u003csup\u003e5\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e Among them, lncRNA ENST00000540293 is differentially expressed in human dental follicle cells and periodontal ligament cells, and is closely related to the osteogenic differentiation of human periodontal ligament cells\u003csup\u003e15,3\u003c/sup\u003e\u003csup\u003e6\u003c/sup\u003e.\u0026nbsp;In this experiment, the calcification of HASMCs induced by DPP4 was used to screen out lncRNAs and mRNAs with obvious differences. The results showed that there were 25 differentially expressed inter-gene lncRNA (8 up-regulated and 17 down-regulated), and the most down-regulated inter-gene lncRNA was ENST00000540293, while lincRNA ENST00000540293 nearby gene was NM-002421, which encoded matrix metalloproteinase-1 (MMP-1).\u0026nbsp;MMP-1 as the prototype of all interstitial collagenases, is also known as collagenase-1, fibroblast collagenase and interstitial collagenase\u003csup\u003e3\u003c/sup\u003e\u003csup\u003e7\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e It is closely related to the regulation of cell microenvironment, and can change cell function to regulate cell differentiation, such as osteogenic differentiation of human periodontal ligament cells and calcification of\u0026nbsp;HASMCs\u003csup\u003e3\u003c/sup\u003e\u003csup\u003e8\u003c/sup\u003e\u003csup\u003e-4\u003c/sup\u003e\u003csup\u003e1\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e\u0026nbsp; \u0026nbsp; \u0026nbsp;\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eIn order to further determine whether the calcification of\u0026nbsp;HASMCs\u0026nbsp;promoted by DPP4 is related to the regulation of lncRNA ENST00000540293. We constructed smart silencer transducers silencing lncRNA ENST00000540293 expression and infected\u0026nbsp;HASMCs, to detect the changes of Runx2, BMP-2, OPG and OPN proteins. The expression of calcification proteins was significantly increased and alizarin red staining also showed a significant increase in calcified nodules after lncRNA ENST0000054029 silenced. The determination of intracellular calcium content and alkaline phosphatase activity showed the same results.Also,\u0026alpha;-SMA which as the surface marker protein of vascular smooth muscle cells also decreased significantly after lncRNA ENST0000054029 gene knockdown.And the immunofluorescence results also showed that the fluorescence intensity of F-actin, myofilament expression and aggregation decreased significantly. It is suggested that DPP4 can induce the decrease of contractile phenotype of\u0026nbsp;HASMCs, which further confirms the tendency of\u0026nbsp;HASMCs\u0026nbsp;to osteoblast.\u003c/p\u003e\n\u003cp\u003eDPP4 is an enzyme that targets the incretin hormone GLP-1 for degradation,is also an important drug target of diabetes and new adipokines, which needs to be cleaved, exfoliated and released from the cell membrane to regulate the circulatory system\u003csup\u003e4\u003c/sup\u003e\u003csup\u003e2\u003c/sup\u003e. MMP-1, MMP-2 and MMP-14 and MMP-9 are involved in the cleavage and shedding of DPP4,and the increased expression of MMP-1 can promote the shedding of DPP4 on the membrane of HASMCs under hypoxia\u003csup\u003e4\u003c/sup\u003e\u003csup\u003e3\u003c/sup\u003e\u003cstrong\u003e.\u003c/strong\u003e DPP4 inhibitor Alogliptin reverses vascular remodelling by downregulating matrix metalloproteinase 1 expression through inhibition of the ERK1/2/NF-\u0026kappa;B signal pathway\u003csup\u003e31\u003c/sup\u003e.The results suggest that DPP4 inhibits the expression of lncRNA ENST00000540293 and MMP-1mRNA. However, inhibiting the expression of\u0026nbsp;lncRNA ENST00000540293 can promote the expression of MMP-1 protein and phosphorylation of ERK1/2 protein.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u0026nbsp; \u0026nbsp; Therefore, we speculate that DPP4 promotes HASMCs calcification by inhibiting lncRNA ENST00000540293, which may be related to the regulation of ERK1/2 signal pathway or MMP-1 protein. In this study, we are unable to investigate all mechanisms of DPP4 silencing lncRNA ENST00000540293 expression and activating ERK1/2 or MMP-1 and their mutual regulation. And Whether DPP4, MMP-1 and lncRNA ENST00000540293 can be used as key targets for the treatment of diabetic vascular calcification remains to be further studied.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eAll authors contributed to the study conception and design. The first draft of the manuscript was written by Tongjien X; material preparation, data collection and analysis were performed by Hao C, Xiongfei X, Xiaolei S, Huqiang H, Hong Z, Sen S and Yong L was responsible for project administration, review, and editing. All authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe study was supported by research grants from the National Natural Science Foundation of China (Fund No.81270358); the Science \u0026amp; Technology Department of Sichuan Province, Grant 2019JY0691. \u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and materials\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data were obtained from the NCBI GEO database (Number :GSE189923).https://www.ncbi.nlm.nih.gov/geo/.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors declare that there is no competing interests regarding the publication of this article.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eJohnson, R. C., Leopold, J. A. \u0026amp; Loscalzo, J. Vascular calcification: pathobiological mechanisms and clinical implications. Circ Res \u003cb\u003e99\u003c/b\u003e, 1044\u0026ndash;1059, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1161/01.RES.0000249379.55535.21\u003c/span\u003e\u003cspan address=\"10.1161/01.RES.0000249379.55535.21\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDemer, L. L. \u0026amp; Tintut, Y. Vascular calcification: pathobiology of a multifaceted disease. Circulation \u003cb\u003e117\u003c/b\u003e, 2938\u0026ndash;2948, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1161/CIRCULATIONAHA.107.743161\u003c/span\u003e\u003cspan address=\"10.1161/CIRCULATIONAHA.107.743161\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eThompson, B. \u0026amp; Towler, D. A. Arterial calcification and bone physiology: role of the bone-vascular axis. Nat Rev Endocrinol \u003cb\u003e8\u003c/b\u003e, 529\u0026ndash;543, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/nrendo.2012.36\u003c/span\u003e\u003cspan address=\"10.1038/nrendo.2012.36\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEddington, H. \u003cem\u003eet al.\u003c/em\u003e Serum phosphate and mortality in patients with chronic kidney disease. Clin J Am Soc Nephrol \u003cb\u003e5\u003c/b\u003e, 2251\u0026ndash;2257, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.2215/CJN.00810110\u003c/span\u003e\u003cspan address=\"10.2215/CJN.00810110\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eXu, J. \u003cem\u003eet al.\u003c/em\u003e Renalase is a novel, soluble monoamine oxidase that regulates cardiac function and blood pressure. J Clin Invest \u003cb\u003e115\u003c/b\u003e, 1275\u0026ndash;1280 (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWronkowitz, N. \u003cem\u003eet al.\u003c/em\u003e Soluble DPP4 induces inflammation and proliferation of human smooth muscle cells via protease-activated receptor 2. Biochim Biophys Acta \u003cb\u003e1842\u003c/b\u003e, 1613\u0026ndash;1621, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bbadis.2014.06.004\u003c/span\u003e\u003cspan address=\"10.1016/j.bbadis.2014.06.004\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eIshibashi, Y., Matsui, T., Maeda, S., Higashimoto, Y. \u0026amp; Yamagishi, S.-i. Advanced glycation end products evoke endothelial cell damage by stimulating soluble dipeptidyl peptidase-4 production and its interaction with mannose 6-phosphate/insulin-like growth factor II receptor. Cardiovascular diabetology \u003cb\u003e12\u003c/b\u003e, 125, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/1475-2840-12-125\u003c/span\u003e\u003cspan address=\"10.1186/1475-2840-12-125\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2013).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLambeir, A.-M., Durinx, C., Scharp\u0026eacute;, S. \u0026amp; De Meester, I. Dipeptidyl-peptidase IV from bench to bedside: an update on structural properties, functions, and clinical aspects of the enzyme DPP IV. Crit Rev Clin Lab Sci \u003cb\u003e40\u003c/b\u003e, 209\u0026ndash;294 (2003).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLee, S. \u003cem\u003eet al.\u003c/em\u003e Dipeptidyl peptidase-4 inhibition to prevent progression of calcific aortic stenosis. Heart (British Cardiac Society) \u003cb\u003e106\u003c/b\u003e, 1824\u0026ndash;1831, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1136/heartjnl-2020-317024\u003c/span\u003e\u003cspan address=\"10.1136/heartjnl-2020-317024\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, Y. \u003cem\u003eet al.\u003c/em\u003e Melatonin ameliorates aortic valve calcification via the regulation of circular RNA CircRIC3/miR-204-5p/DPP4 signaling in valvular interstitial cells. J Pineal Res \u003cb\u003e69\u003c/b\u003e, e12666, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jpi.12666\u003c/span\u003e\u003cspan address=\"10.1111/jpi.12666\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLin, C.-P. \u003cem\u003eet al.\u003c/em\u003e Sitagliptin attenuates arterial calcification by downregulating oxidative stress-induced receptor for advanced glycation end products in LDLR knockout mice. Sci Rep \u003cb\u003e11\u003c/b\u003e, 17851, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/s41598-021-97361-w\u003c/span\u003e\u003cspan address=\"10.1038/s41598-021-97361-w\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2021).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChoi, S.-Y. \u003cem\u003eet al.\u003c/em\u003e Dipeptidyl peptidase-4 inhibitor gemigliptin protects against vascular calcification in an experimental chronic kidney disease and vascular smooth muscle cells. PloS one \u003cb\u003e12\u003c/b\u003e, e0180393, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1371/journal.pone.0180393\u003c/span\u003e\u003cspan address=\"10.1371/journal.pone.0180393\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKapranov, P. \u003cem\u003eet al.\u003c/em\u003e RNA maps reveal new RNA classes and a possible function for pervasive transcription. Science (New York, N.Y.) \u003cb\u003e316\u003c/b\u003e, 1484\u0026ndash;1488 (2007).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGuttman, M. \u0026amp; Rinn, J. L. Modular regulatory principles of large non-coding RNAs. Nature \u003cb\u003e482\u003c/b\u003e, 339\u0026ndash;346, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/nature10887\u003c/span\u003e\u003cspan address=\"10.1038/nature10887\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWu, L. \u003cem\u003eet al.\u003c/em\u003e Comparison of long non\u0026ndash;coding RNA expression profiles in human dental follicle cells and human periodontal ligament cells. Mol Med Rep \u003cb\u003e20\u003c/b\u003e, 939\u0026ndash;950, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3892/mmr.2019.10308\u003c/span\u003e\u003cspan address=\"10.3892/mmr.2019.10308\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2019).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChen, W.-P., Yang, Z.-G., Hu, P.-F., Bao, J.-P. \u0026amp; Wu, L.-D. Acacetin inhibits expression of matrix metalloproteinases via a MAPK-dependent mechanism in fibroblast-like synoviocytes. J Cell Mol Med \u003cb\u003e19\u003c/b\u003e, 1910\u0026ndash;1915, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/jcmm.12564\u003c/span\u003e\u003cspan address=\"10.1111/jcmm.12564\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePurroy, A. \u003cem\u003eet al.\u003c/em\u003e Matrix metalloproteinase-10 deficiency delays atherosclerosis progression and plaque calcification. Atherosclerosis \u003cb\u003e278\u003c/b\u003e, 124\u0026ndash;134, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.atherosclerosis.2018.09.022\u003c/span\u003e\u003cspan address=\"10.1016/j.atherosclerosis.2018.09.022\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eProvenzano, M. \u003cem\u003eet al.\u003c/em\u003e The Association of Matrix Metalloproteinases with Chronic Kidney Disease and Peripheral Vascular Disease: A Light at the End of the Tunnel? \u003cem\u003eBiomolecules\u003c/em\u003e \u003cb\u003e10\u003c/b\u003e, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3390/biom10010154\u003c/span\u003e\u003cspan address=\"10.3390/biom10010154\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWang, P., Xu, J., Wang, Y. \u0026amp; Cao, X. An interferon-independent lncRNA promotes viral replication by modulating cellular metabolism. Science (New York, N.Y.) \u003cb\u003e358\u003c/b\u003e, 1051\u0026ndash;1055, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1126/science.aao0409\u003c/span\u003e\u003cspan address=\"10.1126/science.aao0409\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGiachelli, C. M. Vascular calcification mechanisms. J Am Soc Nephrol \u003cb\u003e15\u003c/b\u003e, 2959\u0026ndash;2964 (2004).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTa, N. N., Li, Y., Schuyler, C. A., Lopes-Virella, M. F. \u0026amp; Huang, Y. DPP-4 (CD26) inhibitor alogliptin inhibits TLR4-mediated ERK activation and ERK-dependent MMP-1 expression by U937 histiocytes. Atherosclerosis \u003cb\u003e213\u003c/b\u003e, 429\u0026ndash;435, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.atherosclerosis.2010.08.064\u003c/span\u003e\u003cspan address=\"10.1016/j.atherosclerosis.2010.08.064\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2010).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLv, J. \u003cem\u003eet al.\u003c/em\u003e Long noncoding RNA H19-derived miR-675 aggravates restenosis by targeting PTEN. Biochem Biophys Res Commun \u003cb\u003e497\u003c/b\u003e, 1154\u0026ndash;1161, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.bbrc.2017.01.011\u003c/span\u003e\u003cspan address=\"10.1016/j.bbrc.2017.01.011\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eBalderman, J. A. F. \u003cem\u003eet al.\u003c/em\u003e Bone morphogenetic protein-2 decreases microRNA-30b and microRNA-30c to promote vascular smooth muscle cell calcification. Journal of the American Heart Association \u003cb\u003e1\u003c/b\u003e, e003905, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1161/JAHA.112.003905\u003c/span\u003e\u003cspan address=\"10.1161/JAHA.112.003905\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2012).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eZhang, L. \u003cem\u003eet al.\u003c/em\u003e H19 knockdown suppresses proliferation and induces apoptosis by regulating miR-148b/WNT/β-catenin in ox-LDL -stimulated vascular smooth muscle cells. J Biomed Sci \u003cb\u003e25\u003c/b\u003e, 11, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s12929-018-0418-4\u003c/span\u003e\u003cspan address=\"10.1186/s12929-018-0418-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMayer, O. [Matrix Gla protein as natural inhibitor of vascular calcification and potential treatment target]. Casopis lekaru ceskych \u003cb\u003e155\u003c/b\u003e, 13\u0026ndash;16 (2016).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eTouyz, R. M. \u003cem\u003eet al.\u003c/em\u003e Vascular smooth muscle contraction in hypertension. Cardiovasc Res \u003cb\u003e114\u003c/b\u003e, 529\u0026ndash;539, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1093/cvr/cvy023\u003c/span\u003e\u003cspan address=\"10.1093/cvr/cvy023\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFragiadaki, M. \u0026amp; Mason, R. M. Epithelial-mesenchymal transition in renal fibrosis - evidence for and against. Int J Exp Pathol \u003cb\u003e92\u003c/b\u003e, 143\u0026ndash;150, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1111/j.1365-2613.2011.00775.x\u003c/span\u003e\u003cspan address=\"10.1111/j.1365-2613.2011.00775.x\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2011).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eEddy, A. A. Overview of the cellular and molecular basis of kidney fibrosis. Kidney Int Suppl (2011) \u003cb\u003e4\u003c/b\u003e, 2\u0026ndash;8 (2014).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR\u0026ouml;hrborn, D., Wronkowitz, N. \u0026amp; Eckel, J. DPP4 in Diabetes. Frontiers in immunology \u003cb\u003e6\u003c/b\u003e, 386, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fimmu.2015.00386\u003c/span\u003e\u003cspan address=\"10.3389/fimmu.2015.00386\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eFan, S.-H., Xiong, Q.-F., Wang, L., Zhang, L.-H. \u0026amp; Shi, Y.-W. Glucagon-like peptide 1 treatment reverses vascular remodelling by downregulating matrix metalloproteinase 1 expression through inhibition of the ERK1/2/NF-κB signalling pathway. Molecular and cellular endocrinology \u003cb\u003e518\u003c/b\u003e, 111005, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.mce.2020.111005\u003c/span\u003e\u003cspan address=\"10.1016/j.mce.2020.111005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCao, X., Yeo, G., Muotri, A. R., Kuwabara, T. \u0026amp; Gage, F. H. Noncoding RNAs in the mammalian central nervous system. Annual review of neuroscience \u003cb\u003e29\u003c/b\u003e (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eMattick, J. S. \u0026amp; Makunin, I. V. Non-coding RNA. \u003cem\u003eHuman molecular genetics\u003c/em\u003e \u003cb\u003e15 Spec No 1\u003c/b\u003e, R17-R29 (2006).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eAmaral, P. P. \u0026amp; Mattick, J. S. Noncoding RNA in development. Mammalian genome: official journal of the International Mammalian Genome Society \u003cb\u003e19\u003c/b\u003e, 454\u0026ndash;492, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1007/s00335-008-9136-7\u003c/span\u003e\u003cspan address=\"10.1007/s00335-008-9136-7\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eCosta, F. F. Non-coding RNAs, epigenetics and complexity. Gene \u003cb\u003e410\u003c/b\u003e, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.gene.2007.12.008\u003c/span\u003e\u003cspan address=\"10.1016/j.gene.2007.12.008\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGhildiyal, M. \u0026amp; Zamore, P. D. Small silencing RNAs: an expanding universe. Nature reviews. Genetics \u003cb\u003e10\u003c/b\u003e, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1038/nrg2504\u003c/span\u003e\u003cspan address=\"10.1038/nrg2504\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eGu, X., Li, M., Jin, Y., Liu, D. \u0026amp; Wei, F. Identification and integrated analysis of differentially expressed lncRNAs and circRNAs reveal the potential ceRNA networks during PDLSC osteogenic differentiation. BMC Genet \u003cb\u003e18\u003c/b\u003e, 100, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/s12863-017-0569-4\u003c/span\u003e\u003cspan address=\"10.1186/s12863-017-0569-4\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2017).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003ePardo, A. \u0026amp; Selman, M. MMP-1: the elder of the family. Int J Biochem Cell Biol \u003cb\u003e37\u003c/b\u003e, 283\u0026ndash;288 (2005).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eHayami, T., Kapila, Y. L. \u0026amp; Kapila, S. MMP-1 (collagenase-1) and MMP-13 (collagenase-3) differentially regulate markers of osteoblastic differentiation in osteogenic cells. Matrix Biol \u003cb\u003e27\u003c/b\u003e, 682\u0026ndash;692, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.matbio.2008.07.005\u003c/span\u003e\u003cspan address=\"10.1016/j.matbio.2008.07.005\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2008).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eLehrke, M. \u003cem\u003eet al.\u003c/em\u003e MMP-1 serum levels predict coronary atherosclerosis in humans. Cardiovascular diabetology \u003cb\u003e8\u003c/b\u003e, 50, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1186/1475-2840-8-50\u003c/span\u003e\u003cspan address=\"10.1186/1475-2840-8-50\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2009).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eKessenbrock, K., Wang, C.-Y. \u0026amp; Werb, Z. Matrix metalloproteinases in stem cell regulation and cancer. Matrix Biol \u003cb\u003e44\u0026ndash;46\u003c/b\u003e, 184\u0026ndash;190, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.matbio.2015.01.022\u003c/span\u003e\u003cspan address=\"10.1016/j.matbio.2015.01.022\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2015).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eNgai, D., Lino, M. \u0026amp; Bendeck, M. P. Cell-Matrix Interactions and Matricrine Signaling in the Pathogenesis of Vascular Calcification. Front Cardiovasc Med \u003cb\u003e5\u003c/b\u003e, 174, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.3389/fcvm.2018.00174\u003c/span\u003e\u003cspan address=\"10.3389/fcvm.2018.00174\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2018).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eDong, C. \u003cem\u003eet al.\u003c/em\u003e Anagliptin stimulates osteoblastic cell differentiation and mineralization. Biomed Pharmacother \u003cb\u003e129\u003c/b\u003e, 109796, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.biopha.2019.109796\u003c/span\u003e\u003cspan address=\"10.1016/j.biopha.2019.109796\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2020).\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eR\u0026ouml;hrborn, D., Eckel, J. \u0026amp; Sell, H. Shedding of dipeptidyl peptidase 4 is mediated by metalloproteases and up-regulated by hypoxia in human adipocytes and smooth muscle cells. FEBS Lett \u003cb\u003e588\u003c/b\u003e, 3870\u0026ndash;3877, doi:\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003e10.1016/j.febslet.2014.08.029\u003c/span\u003e\u003cspan address=\"10.1016/j.febslet.2014.08.029\" targettype=\"DOI\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e (2014).\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"},{"header":"Tables","content":"\u003cp\u003e\u003cstrong\u003eTable 1.\u003c/strong\u003e The characteristics of the lincRNAs induced by DPP4.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003etop 10 downregulated LincRNAs\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd width=\"18.65546218487395%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSeqname\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"9.915966386554622%\"\u003e\n \u003cp\u003e\u003cstrong\u003eChrom\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"15.798319327731093%\"\u003e\n \u003cp\u003e\u003cstrong\u003eGeneSymbol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"12.77310924369748%\"\u003e\n \u003cp\u003e\u003cstrong\u003eFold Change\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eRNA length\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd width=\"14.285714285714286%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNearby Gene Symbol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003eENST00000540293\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003echr11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eAP000619.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e5.4691462\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.018646883\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e1000\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eMMP1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003euc001phf.3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003echr11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eAB231724\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e5.3684649\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.000377659\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e157\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eMMP1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003euc002ywn.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003echr21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eAL109792\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e4.0931103\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.002062516\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e2900\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eKCNJ6\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003eT240875\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003eChr3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eG055454\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e3.9343595\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.003993263\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e8371\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eLYZL4\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003eT240875\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003eChr3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eG055454\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e3.9343595\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.003993263\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e8371\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eVIPR1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003eT046285\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003eChr10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eG010780\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e3.4726155\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.012499379\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e1603\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eCEP55\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003eENST00000412427\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003eChr1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eRP11-380J14.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e3.3043558\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.002605688\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e4676\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eATP13A2\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003eENST00000452326\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003eChr22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eAC000067.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e3.2458684\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.012123994\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e287\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eC22orf29\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003eENST00000452326\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003eChr22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eAC000067.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e3.2458684\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.012123994\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e287\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eCDC45\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.65546218487395%\"\u003e\n \u003cp\u003euc002xuq.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"9.915966386554622%\"\u003e\n \u003cp\u003eChr20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.798319327731093%\"\u003e\n \u003cp\u003eAK055386\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"12.77310924369748%\"\u003e\n \u003cp\u003e2.9207398\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e0.008848319\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003e3404\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.285714285714286%\"\u003e\n \u003cp\u003eKCNB1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003etop 8 upregulated LincRNAs\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003e\u003cstrong\u003eSeqname\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003e\u003cstrong\u003eChrom\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003e\u003cstrong\u003eGene Symbol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e\u003cstrong\u003eFold Change\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e\u003cstrong\u003eP-value\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e\u003cstrong\u003eRNA length\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e\u003cstrong\u003eNearby Gene Symbol\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eENST00000450216\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eCTA-150C2.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e4.1538977\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.033626171\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e678\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eAPOBEC3B\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eENST00000450216\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr22\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eCTA-150C2.13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e4.1538977\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.033626171\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e678\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eAPOBEC3A\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eENST00000607314\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eRP11-1002K11.1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e3.3245744\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.000171600\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e1835\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eNRG1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eNR_003133\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eGBP1P1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e3.035663\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.017197101\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e1234\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eLRRC8B\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eENST00000520544\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr8\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eRP11-760H22.2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e2.2794756\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.006961939\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e490\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eCOL14A1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eENST00000562082\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr10\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eRP11-119F7.5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e2.0858913\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.009993759\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e2295\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eSTOX1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eT365864\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr9\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eG086505\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e2.0357136\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.033876979\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e1819\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eTRAF1\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"19.12751677852349%\"\u003e\n \u003cp\u003eENST00000608605\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"9.395973154362416%\"\u003e\n \u003cp\u003eChr3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"17.449664429530202%\"\u003e\n \u003cp\u003eRP11-804H8.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"11.74496644295302%\"\u003e\n \u003cp\u003e2.0171033\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"13.758389261744966%\"\u003e\n \u003cp\u003e0.006191725\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003e2060\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"14.261744966442953%\"\u003e\n \u003cp\u003eCISH\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 2\u003c/strong\u003e.The specific primers of LncRNA ENST00000540293,MMP-1 and\u0026beta;-actin were designed.\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"23.65771812080537%\"\u003e\n \u003cp\u003eGene name\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd colspan=\"2\" valign=\"top\" width=\"76.34228187919463%\"\u003e\n \u003cp\u003ePrimers \u0026nbsp;sequence\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"23.65771812080537%\"\u003e\n \u003cp\u003eLncRNA ENST00000540293\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.604026845637584%\"\u003e\n \u003cp\u003eForward\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"60.738255033557046%\"\u003e\n \u003cp\u003e5\u0026apos;-CTGGACCAGGTATCGGAGGAGATG-3\u0026apos;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"20.439560439560438%\"\u003e\n \u003cp\u003eReverse\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"79.56043956043956%\"\u003e\n \u003cp\u003e5\u0026apos;-GGTAGCCACTCAGCAGTTGATCC-3\u0026apos;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"23.65771812080537%\"\u003e\n \u003cp\u003eMMP-1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.604026845637584%\"\u003e\n \u003cp\u003eForward\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"60.738255033557046%\"\u003e\n \u003cp\u003e5\u0026apos;-AGATTCTACATGCGCACAAATC-3\u0026apos;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"20.439560439560438%\"\u003e\n \u003cp\u003eReverse\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"79.56043956043956%\"\u003e\n \u003cp\u003e5\u0026apos;-CCTTTGAAAACCGGACTTCAT-3\u0026apos;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd rowspan=\"2\" valign=\"top\" width=\"23.65771812080537%\"\u003e\n \u003cp\u003e\u0026beta;-actin\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"15.604026845637584%\"\u003e\n \u003cp\u003eForward\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"60.738255033557046%\"\u003e\n \u003cp\u003e5\u0026apos;-CCTGGCACCCAGCACAAT-3\u0026apos;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"top\" width=\"20.439560439560438%\"\u003e\n \u003cp\u003eReverse\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"top\" width=\"79.56043956043956%\"\u003e\n \u003cp\u003e5\u0026apos;-GCCGATCCACACGGAGTA-3\u0026apos;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003eTable 3.\u0026nbsp;\u003c/strong\u003eThe characteristics of the mRNAs induced by DPP4\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003etop 10 downregulated mRNAs\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eSeqname\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003eChrom\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eRNA \u0026nbsp;length\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003eFold Change\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eGene Symbol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003etype\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_144634\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e916\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e7.7044098\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.001159771\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eLYZL4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_198404\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr13\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e2133\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e5.2321373\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.003622246\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eKCTD4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_003280\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e705\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e5.1611148\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.002562575\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eTNNC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_002240\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr21\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e2790\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e4.9792082\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.008725652\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eKCNJ6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_005582\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr5\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e2725\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e4.8654311\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.003843377\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eCD180\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_003783\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e3548\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e4.7786871\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.001342022\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eB3GALT2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_145060\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr18\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e2893\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e4.6475968\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.011197155\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eSKA1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_021158\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e2554\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e4.5764374\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.011266314\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eTRIB3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_002421\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr11\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e2081\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e4.2991079\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.028861407\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003eMMP1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"17.843866171003718%\"\u003e\n \u003cp\u003eNM_002849\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.736059479553903%\"\u003e\n \u003cp\u003echr12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e3516\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.799256505576208%\"\u003e\n \u003cp\u003e4.2792469\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003e0.004702521\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"14.12639405204461%\"\u003e\n \u003cp\u003ePTPRR\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.241635687732343%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e\n\u003cp\u003e\u003cstrong\u003etop 10 upregulated mRNAs\u003c/strong\u003e\u003c/p\u003e\n\u003ctable border=\"1\" cellpadding=\"0\" cellspacing=\"0\" width=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.367346938775512%\"\u003e\n \u003cp\u003eSeqname\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.719851576994435%\"\u003e\n \u003cp\u003eChrom\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.358070500927644%\"\u003e\n \u003cp\u003eRNA \u0026nbsp;length\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.769944341372913%\"\u003e\n \u003cp\u003eFold Change\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.213358070500927%\"\u003e\n \u003cp\u003eP-value\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.543599257884972%\"\u003e\n \u003cp\u003eGene Symbol\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.027829313543599%\"\u003e\n \u003cp\u003etype\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.367346938775512%\"\u003e\n \u003cp\u003eNM_000064\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.719851576994435%\"\u003e\n \u003cp\u003echr19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.358070500927644%\"\u003e\n \u003cp\u003e5148\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.769944341372913%\"\u003e\n \u003cp\u003e27.6095442\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.213358070500927%\"\u003e\n \u003cp\u003e0.002442785\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.543599257884972%\"\u003e\n \u003cp\u003eC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.027829313543599%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.367346938775512%\"\u003e\n \u003cp\u003eNM_001080400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.719851576994435%\"\u003e\n \u003cp\u003echr19\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.358070500927644%\"\u003e\n \u003cp\u003e6353\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.769944341372913%\"\u003e\n \u003cp\u003e11.528119\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.213358070500927%\"\u003e\n \u003cp\u003e0.001785257\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.543599257884972%\"\u003e\n \u003cp\u003ePLIN4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.027829313543599%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.367346938775512%\"\u003e\n \u003cp\u003eNM_000668\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.719851576994435%\"\u003e\n \u003cp\u003echr4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.358070500927644%\"\u003e\n \u003cp\u003e2707\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.769944341372913%\"\u003e\n \u003cp\u003e11.3082146\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.213358070500927%\"\u003e\n \u003cp\u003e0.015208381\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.543599257884972%\"\u003e\n \u003cp\u003eADH1B\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.027829313543599%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.367346938775512%\"\u003e\n \u003cp\u003eNM_000014\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"8.719851576994435%\"\u003e\n \u003cp\u003echr12\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.358070500927644%\"\u003e\n \u003cp\u003e4678\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.769944341372913%\"\u003e\n \u003cp\u003e11.2869199\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.213358070500927%\"\u003e\n \u003cp\u003e0.000195162\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"13.543599257884972%\"\u003e\n \u003cp\u003eA2M\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" width=\"15.027829313543599%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\" width=\"18.367346938775512%\"\u003e\n \u003cp\u003eNM_000669\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\" 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valign=\"bottom\" width=\"15.027829313543599%\"\u003e\n \u003cp\u003eprotein_coding\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n\u003c/table\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Dipeptidyl peptidase 4, Vascular Calcification, RNA Chip, Long non-coding RNA","lastPublishedDoi":"10.21203/rs.3.rs-1656423/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-1656423/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eArterial media calcification caused by diabetes is an important cause of cardiovascular calcification. Dipeptidyl peptidase-4 (DPP4) is associated with diabetic arterial media calcification. While, long non-coding RNA(lncRNA) is closely related to the cardiovascular diseases and calcification caused by diabetes. Experiment\u0026nbsp;have shown that LncRNA can regulate DPP4 activation to modulate senescence and lncRNA ENST00000540293 may associated with human periodontal ligament cells development. our previous studies showed that DPP4 can promote the calcification of human aortic smooth muscle cells (HASMCs) by activating the ERK/NF-kB pathway, but it is unknown whether it is related to the regulation of lncRNA. Calcification of HASMCs was induced by DPP4. There was a significant difference in the expression of lncRNAs and mRNAs between normal and calcified cells detected by gene chip technology. Based on the results of microarray detection, we found that lncRNA ENST00000540293 may be involved in vascular calcification induced by DPP4 through regulating target genes.\u003c/p\u003e","manuscriptTitle":"Expression profile of lncRNA induced by DPP4 in human aortic smooth muscle cells and calcification gene prediction","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2022-06-03 22:21:49","doi":"10.21203/rs.3.rs-1656423/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"0eb4664b-d286-4fa1-a918-f28b0b6a8ed2","owner":[],"postedDate":"June 3rd, 2022","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2022-07-22T07:14:28+00:00","versionOfRecord":[],"versionCreatedAt":"2022-06-03 22:21:49","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-1656423","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-1656423","identity":"rs-1656423","version":["v1"]},"buildId":"rHA-KDH7Qsr4HCuvH75dn","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}