{"paper_id":"449ffdd2-1391-421d-9a46-71be79e19ed0","body_text":"The causative effect of CXCR7 on experimental autoimmune prostatitis injury and fibrosis | 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 The causative effect of CXCR7 on experimental autoimmune prostatitis injury and fibrosis Yi zhang, Rui Feng, Weikang Wu, Xianhong Liu, Cong Huang, Xianguo Chen, and 2 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4186657/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 Chronic prostatitis and Pelvic Pain syndrome (CP/CPPS) is an autoimmune inflammatory disease characterized by pelvic or perineal pain and infiltration of inflammatory cells in the prostate. C-X-C chemokine receptor type 7 (CXCR7), also known as the atypical chemokine receptor 3 (ACKR3) receptor, are atypical chemokine receptors. Having been shown to play a key role in inflammatory processes, whether CXCR7 influences the role of autoimmune prostate and immune regulation and its mechanism of action are unclear. In this study, a mouse model of experimental autoimmune prostatitis was constructed by subcutaneous injection of antigen, and CXCR7 agonist was administered to investigate the effects of CXCR7 on the proportion of immune cells and fibrosis in CP/CPPS. Western blotting, immunohistochemical staining and Immunofluorescence, flow cytometry, and masson staining were used to study the possible regulatory mechanisms. CXCR7 agonists can significantly reduce pain and prostatic inflammation, and in vivo flow studies have shown that they affect the TH17/Treg cell ratio. To elucidate the potential mechanisms by which CXCR7 influences the pathogenesis of CNP, we conducted simultaneous RNA-seq and non-targeted metabolome sequencing. Our findings suggest that CXCR7 agonists alleviate fibrosis in autoimmune prostatitis by inhibiting the TGFβ/SMAD pathway. This study provides a valuable immunological basis for CNP to intervene CP/CPPS therapy with CXCR7 as the target. chronic prostatitis CXCR7 inflammation fibrosis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Figure 7 Introduction Chronic prostatitis/Chronic pelvic pain syndrome (CP/CPPS) is one of the most common diseases of the urogenital system in young and middle-aged men, and is also listed as NIH Class III prostatitis, which is considered to be the most common type. The incidence is about 13.8% [1, 2].CP/CPPS is a disease that seriously affects the daily life of young and middle-aged men, and is characterized by pelvic perineal pain, obstructive or irritating lower urinary tract urination symptoms, and sexual dysfunction. The pathogenesis of CP/CPPS is complex, including bacterial infection, diet, immune cell infiltration, endocrine abnormalities [3] and Autonomic Nervous System Dysfunction [4]. However, the pathogenesis of CP/CPPS has not been clarified yet, so the therapeutic effect is not satisfactory. Therefore, it is necessary and necessary to find new therapeutic targets for CP/CPPS and explore new therapeutic methods. Although the role of CXCL12-CXCR4 signaling has been well studied, CXCR4 has high clinical value in hematopoietic stem cell and cancer biology, as well as in inflammatory diseases [5–8]. However, when chemokines participate in the inflammatory process, CXCR7, as a non-classical chemokine receptor, is up-regulated in many inflammatory diseases, such as rheumatoid arthritis [9], multiple sclerosis [10], atherosclerosis [11], and septic peritonitis [12]. However, unlike classical G protein-coupled receptors (GPCRS), CXCR7 does not signal through the G protein pathway, but instead the main ligands are the chemokines CXCL11 and CXCL12. Although CXCL12 has a greater affinity for CXCR7 than CXCR4, its receptor function is closely related to CXCR4. Depending on the cell type, CXCR4 either performs similar functional control in combination with CXCL12 or interacts with CXCR4 in quite complex and diverse ways [13, 14]. During the progression of chronic inflammatory diseases, fibroblasts are activated to repair tissues abnormally, and tissue fibrosis is generally the end result of chronic inflammatory response, mainly the production of excessive collagen Abnormal activation of fibers and fibroblasts leads to organ dysfunction. In addition to smooth muscle cells, blood vessels and nerves, the prostate stroma is basically composed of collagen fibers, elastic fibers and undifferentiated fibroblasts [15]. Under the action of cytokines and chemokines (CXCL5, CXCL8, and CXCL12), intraprostatic myofibroblasts activate and secrete α-SMA at the site of inflammation [16], and may lead to permanent fibrosis in the subsequent progression of pathological processes. However, the effect of CXCR7 on prostatic inflammation and fibrosis in CP/CPPS remains unclear. In addition, CXCR7 agonists improved post-EAP mouse fibrosis by altering the spleen TH17/Treg cell ratio, and it is noteworthy that these conclusions are based on systematic inhibition of CXCR7 and CXCR4, making it an attractive target for therapeutic intervention. In this study, we investigated the effects of CXCR7 on inflammation and the specific mechanisms that regulate fibrosis in EAP mouse models. Our results show that in mice treated with CXCR7 agonists, disease severity and inflammatory cell infiltration are reduced, and the progression of prostate fibrosis is alleviated through the TGFβ/SMAD pathway, reducing collagen accumulation. Materials and Methods Experimental mice and EAP Induction We purchased male Sprague-Dawley rats from the Experimental Animal Center of Anhui Medical University for experiments. Prostate antigen preparation. Healthy male NOD/LtJ (NOD/ShiLtJ, 4 weeks old) mice. All mice, CNP Model Animal Research Center, Nanjing University (Nanjing).We domesticated them for a week before the experiment. All operations were performed under anesthesia. Chloral hydrate was used that the purpose is to reduce pain. Before the experiment began, all the mice underwent a week of acclimatization to the laboratory environment. All animals were kept in a dedicated pathogen free research animal facility at Anhui Medical University. In short, prostate homogenates were obtained from Sprague - Dawley rats and emulsified with superclear and equal amounts of full Freund adjuvant. The mice were immunized on day 0 and day 28 respectively according to the experimental plan. Then the experimental mice were injected with VUF11027(5mg/kg/ day, DMSO as a solvent) through the tail vein for two consecutive weeks. The mice were killed on day 50. Hematoxylin-Eosin Staining Prostate tissue was taken, fixed with 10% formalin solution, paraffin embedded, sliced, dewaxed to water, stained with hematoxylin dye, then dyed with eosin dye, and finally dehydrated and sealed. The degree of inflammation was quantified on a 4-point scale from 0 to 3 [ 17 ], with the following criteria: 0, no inflammation; 1, mild inflammation, and clear monocytes around the blood vessels; 2, moderate inflammation, moderate perivascular monocytes surrounding; 3. Severe inflammation, bleeding, and a large number of monocytes in parenchyma with obvious infiltration around blood vessels. IHC Paraffin embedded tissue sections were dewaxed to water, heated at PH = 6.0, 0.01M sodium citrate solution at 100° for 15 minutes for antigen repair, and then put into 3% hydrogen peroxide solution to block endogenous peroxidase, and washed with PBS buffer three times for five minutes each time. Then it was sealed with 10% fetal bovine serum. The corresponding primary resistance was added and incubated at 4 degrees overnight. After washing with PBS for three times, the secondary antibody was added, incubated at room temperature for 2 hours, tested with freshly configured DAB chromogenic solution, and finally restained the nucleus with hematoxylin. The expression level was quantitatively analyzed using image J software Masson staining Paraffin-embedded tissue sections were dewaxed to water, immersed in 2.5% potassium dichromate mordant overnight, and rinsed. The slices are then put into Weigert iron hematoxylin dye for 1min, then rinsed, and hematoxylin differentiation solution for 10sec. After rinsing with running water, the slices are placed into ponceau acid fuchsin dye solution for 10min. After rinsing with distilled water, the slices are placed into 1% phosphomolybdic acid solution for 3min. After rinsing and drying slightly, the slices are placed into 2.5% aniline blue solution for 1min and then rinsed and differentiated with 1% acetic acid. Dehydration with anhydrous ethanol three times, xylene transparent, neutral resin seal. image J software was used to quantitatively analyze the proportion of collagen fibers. Sirius Red staining Paraffin-embedded tissue sections were dewaxed to water, immersed in Sirius red staining for 10 minutes, dehydrated with anhydrous ethanol three times, xylene transparent section for 5 minutes, and sealed with neutral resin. image J software was used to quantitatively analyze the proportion of collagen fibers. Immunofluorescence The paraffin sections were dewaxed to water, followed by antigen repair, sealing, adding primary antibody (CXCR7, ab72100, Abcam), and incubated in a wet box at 4° overnight. After adding the secondary antibodies (IL17A, ab79056, Abcam), the cells were incubated for 90 minutes away from light, DAPI was re-dyed in the nucleus, and the cells were incubated at dark room temperature. The co-localization of CXCR7 and IL17A was confirmed by Olympus FV3000 confocal laser scanning microscope. Western Blotting Assays The protein extract was separated by 12.5% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membrane. Incubate with 5% milk for 2 hours, then incubate with the corresponding primary antibody at 4°C overnight. GAPDH was used as loading control. TBST washing membrane was incubated with the corresponding secondary antibody (anti-rabbit or anti-mouse IgG, 1:50,000, Elabscience) for 2 hours, and then washed with TBST. The membranes were washed, incubated with secondary antibodies for 2 h, and visualized using an EZ-ECL Kit (Biological Industries, Israel). Mouse Spleen Dissociation and Flow Cytometry The proportion of Th17 and Treg cells was measured by flow cytometry. The splenic single-cell suspension was prepared according to our previous method [ 18 ]. Fitc-labeled CD4(BD Pharmingen, 557307,USA) and APC-labeled CD3e(BD Pharmingen, 553066,USA) were incubated with surface antibodies to APC-labeled CD25(BD Pharmingen, 553075,USA). After washing PBS again, Add 1 ml per tube containing phorbol 12myristate 13-acetate (PMA, MultiSciences, China), lonomycin (MultiSciences, China) 1640 medium (Gibco, USA) from monenin (MultiSciences, China) and Monenin (Multisciences, China) were incubated at 37℃ for 4 h and washed again. The cells were then sequentially immobilized and infiltrated, and incubated at 4 ° C for 60 min with intracellular antibodies, including PE-labeled IL-17A(BD Pharmingen, 559502, USA) and PE-labeled FoxP3(BD Pharmingen, 560401, USA). After washing again, analysis was performed with FACS Calibur flow cytometry (Beckman Coulter, USA). The resulting data is processed by FlowJo software (Tree Star, Ashland). mRNA Sequencing Experimental TRIzol reagent was used to extract the total RNA of the sample. Smarter amplification kit was used to reverse transcribed the qualified total RNA and amplified the cDNA. The qualified cDNA was constructed by Tn5 transposition enzyme method: First, when the cDNA was randomly interrupted by transposase, the interrupted fragment was added with sequencing splices. Then, part of the sequencing splices were used as primers to amplify and enrich the library. After the amplified library was purified by magnetic beads and qualified by Agilent 2100 Bioanalyzer, Sequencing with Illumina Nova6000 other sequencers produced a double end number of 150bp. DESeq2 package was used to analyze the differentially expressed genes, which were consistent with P-value &LT. Genes with a threshold of 0.05 and |LogFC2| > 1 were defined as differentially expressed genes (DEGs). Untargeted Metabolomics Analysis Weigh the sample into a 1.5mL centrifuge tube, add two small steel balls, methanol-water (V:V = 4:1, containing L-2-chlorophenylalanine, 4 µg/mL); Put it in the refrigerator at -40℃ for 2 min, and then put it into the grinding machine (60 Hz, 2 min). Ultrasonic extraction in ice water bath for 10 min; Stand at -40℃ for 30 min; Centrifuge at low temperature for 10 min (12000 rpm, 4℃), and put the supernatant into a glass derived bottle; Use a centrifugal concentrator to dry the sample. Methoxyamine hydrochloride pyridine solution (15 mg/mL) was added to a glass derived vial and incubated at 37°C for 60 min for oxime reaction. Remove samples after add 50 ul BSTFA derivatization reagent and 20 mu L n-hexane, add 10 kinds of internal standard 10 ul, 60 min at 70 ℃ reaction; After the samples were removed, they were placed at room temperature for 30 min for GC-MS analysis. The analytical instrument of the experiment was GMS-7890B /5977A (Aglient, USA). Weigh the sample into a 1.5mL EP tube, add two small steel balls, and add methanol-water (V:V = 4:1, containing L-2-chlorophenylalanine, 4 µg/mL); After pre-cooling for 2 min in the − 40℃ refrigerator, put it into the grinder for grinding (60 Hz, 2 min); Ultrasonic extraction in ice water bath for 10 min and standing at -40℃ for 2 hours; Centrifuge for 10 min (12000 rpm, 4℃), the supernatant absorbed by syringe was filtered by 0.22 µm organic phase pinhole filter, transferred to LC sample vial and stored at -80℃ until LC-MS analysis. Quality control sample (QC) is prepared by mixing the extract of all samples in equal volume. The analytical instrument for the experiment was ACQUITY UPLC I-Class system (Waters Corporation, USA). Statistical Analysis All results were expressed as the mean ± standard deviation (SD). GraphPad Prism version 8.0 software and (GraphPad Software, San Diego, CA).SPSS software version 26.0 (IBM Corp, Armonk, NY, USA) was used for statistical analysis. Two-tailed Student’s t-tests or Wilcoxon rank sum test was applied for the comparison of two groups. Results were considered statistically significant if p-value < 0.05. Results CXCR7 agonists relieved inflammation in EAP mice Following the procedure shown in Fig. 1A, mice were successfully modeled using a mixture of Pags and full Fredringer adjuvant, followed by a 14-day caudal intravenous injection of CXCR7 agonist. Mechanical stimulation of the pelvic area showed that the response frequency of EAP mice was significantly higher than that of the normal group, while the response frequency after CXCR7 agonist was lower than that of the control group, and the response frequency was correlated with the applied force (Fig. 1B). HE histological section (Fig. 1C) showed that mice in EAP group had more inflammatory cell infiltration than mice in Control group, indicating that EAP mice were successfully induced. However, histopathological scores (Fig. 1D) showed that inflammation was reduced after the administration of CXCR7 agonists compared to the control group who received tail vein DMSO. Chronic inflammation of prostate is closely related to cell necrosis and apoptosis, which has been confirmed in multiple experiments [ 19 – 22 ]. However, CXCR7 is involved in the process of apoptosis [ 23 , 24 ], and it is obvious from immune tissue Caspase3 staining (Fig. 1E) that the EAP group has more apoptosis than the Control group. The results of CXCR7 agonist and DMSO control group indicate that CXCR7 may also be involved in apoptosis in prostatitis. CXCR7 aggravates prostatitis by affecting TH17/Treg cell imbalance HE staining (Fig. 1B) and immunohistochemical CD45 staining (Fig. 2A) confirmed the increase of infiltrating inflammatory cells in the prostate of EAP mice. IL17A is highly expressed in the serum of prostatitis mice and has anti-inflammatory function [ 25 ], which was also proved by ELISA experiments. CXCR7 can also reduce the expression of IL17A (Fig. 2B). We speculated that CXCR7 has different expression of TH17 cells and Treg cells in prostatitis. Therefore, we conducted co-localization of CXCR7 and TH17 cells respectively (Fig. 2C). Compared with the control group,CXCR7 agonist group showed that CXCR7 + IL17A + cells could be reduced. Flow cytometry results showed that in the CXCR7 agonist group, the differentiation percentage of TH17 cells in spleen decreased (Fig. 3A,C), while the differentiation percentage of Treg cells increased (Fig. 3B,D). Taken together, these observations suggest that CXCR7 is involved in prostatic inflammation and that CXCR7 agonists restore EAP-induced Th17/Treg imbalances. RNA-seq in EAP mice In order to investigate the mechanism of CXCR7 on the regulation of EAP mice, we divided the mice into EAP + VUF11207 group and EAP + DMSO group, respectively, and conducted rna sequencing analysis and off-target metabolic sequencing. By p&lt; 0.05 and |logFC|&gt; 1 is the cutoff value. Figure 4A shows the heat map of the expression of differential genes in the two groups, while Fig. 4B volcano map shows that 144 genes are up-regulated and 97 genes are down-regulated after CXCR7 agonist is used compared with the control group. Subsequently, Fig. 4C and Fig. 4D showed the pathway diagram of GO and KEGG enrichment analysis of differential genes, which can be seen as endopeptidase activity, serine − type endopeptidase activity, serine − type peptidase activity, serine hydrolase activity, etc. Matrix metalloproteinase is a classic hydrolase with endopeptidase activity, which can be secreted into extracellular matrix and deeply participate in the fibrosis process of organs [ 26 , 27 ]. untargeted metabolomics in EAP mice Because we wanted to find out which metabolic processes and products regulated by CXCR7 in chronic prostatitis are involved in the occurrence and development of the disease, we conducted non-targeted metabolites detection on prostate tissues of EAP + VUF11207 group and EAP + DMSO group. In order to ensure more complete and comprehensive detection data, We used both LC-MS and GC-MS platforms for detection. In order to screen for differential metabolites, as shown in Fig. 5A and Fig. 5B, GC-MS and LC-MS products respectively, in which the red origin represents significantly up-regulated metabolites in the experimental group, the blue origin represents significantly down-regulated metabolites, and the gray dots represent insignificant metabolites. Correlation network (cornetwork) was used to calculate the correlation between differential metabolites and differential metabolite response intensity data based on pearson correlation analysis. pvalue &lt was selected. 0.05,correlation &gt; 0.95 relationship pairs are plotted in the network diagram (Fig. 5C). Correlation analysis can help measure the degree of correlation between metabolites with significant differences, and further understand the interrelationship between metabolites in the process of biological state change. Correlation analysis used Pearson correlation coefficient to measure the degree of linear correlation between the two metabolites (Fig. 5D). The TOP20 differential metabolic pathways were shown through the KEGG pathway mapper function, and the differential metabolites were shown according to the up-down-regulation information (Fig. 5E). Notably, after CXCR7 agonist use, the Glycolysis/Gluconeogenesis pathway and Alanine, aspartate and glutamate metabolism pathway were identified in enrichment analysis. Glutamic acid is involved in the metabolic process of glutathione, which is considered to be a contributing factor to the mechanism of the pathological process of fibrosis in many pathological conditions [ 28 ]. The involvement of glycolysis has been pointed out by many studies to be involved in the pathological process of fibrosis in various organs, such as lung [ 29 , 30 ], heart [ 31 ], kidney [ 32 , 33 ], liver [ 34 , 35 ], etc. CXCR7 affects chronic prostatitis fibrosis To test whether CXCR7 affects prostatitis fibrosis, we administered CXCR7 agonists to EAP mice in a caudal vein. Firstly, Masson's (Fig. 6A), Sirius Red (Fig. 5B) and alfa-smooth muscle actin (α-SMA) (Fig. 6C) staining were used to verify that EAP mice had fibrosis compared with normal mice. This was consistent with the results of other researchers [ 36 – 38 ], but it was also observed that the treatment of CXCR7 agonists significantly improved the fibrosis of prostate tissue compared with the treatment of DMSO controls. Our experimental results (Fig. 6D-E) suggest that CXCR7 has an effect on chronic prostatitis fibrosis. These results suggest that CXCR7 is influencing prostatitis fibrotic process. To further identify CXCR7-mediated pathways of fibrotic processes. We found that VUF11207 improved the expression of TGFβ and decreased the phosphorylated protein level of SMAD2 (Fig. 7A-C). At the protein level, the classic signature proteins of fibrosis, α-SMA (Fig. 7A,D) and col1a1 (Fig. 7A,E), were also significantly reduced after being treated with CXCR7 agonists. These results suggest an important role of CXCR7-mediated pathway in the progression of prostatitis fibrosis. Discussion Our previous findings determined that the CXCL12/ CXCR4 axis is involved in the pathogenesis and development of CNP by regulating the infiltration of CD4 + T cells into the prostate gland and forming inflammatory nests [ 39 ]. However, the downstream ligands of CXCL12 are CXCR4 and CXCR7. Therefore, this study focused on the use of CXCR7 agonists. CXCR7 belongs to the superfamily of 7 transmembrane G-protein-coupled receptors, but cannot stimulate the typical G-protein-dependent pathway, but is involved in various pathophysiological functions. Recent studies have shown that the effects of CXCR7 on inflammation can be observed in a variety of diseases. Including cancer [ 40 – 42 ], bone and joint inflammation [ 43 ], atherosclerotic thrombosis [ 11 ] and inflammatory demyelinating diseases [ 10 ]. Fibrosis is a hallmark of various inflammatory processes. Many previous studies have fully demonstrated that CXCR7 is fully involved in the fibrosis process in the disease. The differential expression of SDF-1, CXCR7 and CXCR4 in hepatic sinusoidal endothelial cells leads to fibrotic transformation of vascular niches. selective CXCR7 activation in liver sinusoidal endothelial cells abrogated fibrogenesis[ 44 ]. Administration of CXCR7 agonists after lung injury can inhibit β-catenin-dependent Jag1 induction in injured PCEC[ 45 ]. CXCR7 mainly mediates regeneration, repair and proliferation of endothelial cells, and alveolar hypoxia can increase the expression of CXCR7 in lung endothelium [ 46 ]. In heart disease, CXCL12 is mainly expressed in vascular smooth muscle cells, and smooth muscle specific CXCL12 KO mutation leads to severe heart hypertrophy, and significant specific activation of pERK signal after CXCR7 agonist can alleviate the progression of myocardial hypertrophy in mice [ 47 ]. In conclusion, CXCR7 can influence fibrotic progression in inflammatory diseases. In this study, we found that CXCR7 in a mouse model of prostatitis affects the progression of chronic prostatitis and the development of TH17/Treg cell ratio in the model mice, and the fibrosis experiment further demonstrated that the functional level of CXCR7 affects the pathogenesis of chronic prostatitis fibrosis. In terms of mechanism, CXCL12/CXCR7 can regulate TGFβ-SMAD2/3 signaling pathway to promote the onset of chronic prostatitis and the progression of fibrosis. At present, the main drugs to treat chronic prostatitis are analgesics and proprietary Chinese medicine, and there is no effective treatment. Together, our results suggest that CXCR7 may be a promising therapeutic target for chronic prostatitis. Declarations Conflicts of interest disclosure None. Availability of Data and Materials None. Funding support The National Natural Science Foundation of China 82170787, 82370776, and 82300872. Author contribution LZ, XC and CJ: conception and design this study; ZY, FR and WWK: collection and assembly of data; WWK, HC, ZL,CJ: data Analysis and interpretation; ZY and FR: manuscript writing. Final Approval of Manuscript: All the authors. Acknowledgments We wish to thank the Center for Scientific Research of the First Affiliated Hospital of Anhui Medical University for valuable help in our experiments. References Krieger JN, Nyberg L Jr., Nickel JC. NIH consensus definition and classification of prostatitis. JAMA. 1999;282(3):236–7. Bartoletti R, Cai T, Mondaini N, Dinelli N, Pinzi N, Pavone C, Gontero P, Gavazzi A, Giubilei G, Prezioso D, et al. Prevalence, incidence estimation, risk factors and characterization of chronic prostatitis/chronic pelvic pain syndrome in urological hospital outpatients in Italy: results of a multicenter case-control observational study. J Urol. 2007;178(6):2411–5. discussion 2415. Strauss AC, Dimitrakov JD. New treatments for chronic prostatitis/chronic pelvic pain syndrome. Nat Rev Urol. 2010;7(3):127–35. He H, Luo H, Qian B, Xu H, Zhang G, Zou X, Zou J. Autonomic Nervous System Dysfunction Is Related to Chronic Prostatitis/Chronic Pelvic Pain Syndrome. World J Mens Health 2023. Daniel SK, Seo YD, Pillarisetty VG. The CXCL12-CXCR4/CXCR7 axis as a mechanism of immune resistance in gastrointestinal malignancies. Semin Cancer Biol. 2020;65:176–88. Gupta N, Mohan CD, Shanmugam MK, Jung YY, Chinnathambi A, Alharbi SA, Ashrafizadeh M, Mahale M, Bender A, Kumar AP et al. CXCR4 expression is elevated in TNBC patient derived samples and Z-guggulsterone abrogates tumor progression by targeting CXCL12/CXCR4 signaling axis in mice model. Environ Res 2023:116335. Heidegger I, Fotakis G, Offermann A, Goveia J, Daum S, Salcher S, Noureen A, Timmer-Bosscha H, Schafer G, Walenkamp A, et al. Comprehensive characterization of the prostate tumor microenvironment identifies CXCR4/CXCL12 crosstalk as a novel antiangiogenic therapeutic target in prostate cancer. Mol Cancer. 2022;21(1):132. Gallego C, Vetillard M, Calmette J, Roriz M, Marin-Esteban V, Evrard M, Aknin ML, Pionnier N, Lefrancois M, Mercier-Nome F, et al. CXCR4 signaling controls dendritic cell location and activation at steady state and in inflammation. Blood. 2021;137(20):2770–84. Watanabe K, Penfold ME, Matsuda A, Ohyanagi N, Kaneko K, Miyabe Y, Matsumoto K, Schall TJ, Miyasaka N, Nanki T. Pathogenic role of CXCR7 in rheumatoid arthritis. Arthritis Rheum. 2010;62(11):3211–20. Pouzol L, Baumlin N, Sassi A, Tunis M, Marrie J, Vezzali E, Farine H, Mentzel U, Martinic MM. ACT-1004-1239, a first-in-class CXCR7 antagonist with both immunomodulatory and promyelinating effects for the treatment of inflammatory demyelinating diseases. FASEB J. 2021;35(3):e21431. Cebo M, Dittrich K, Fu X, Manke MC, Emschermann F, Rheinlaender J, von Eysmondt H, Ferreiros N, Sudman J, Witte A, et al. Platelet ACKR3/CXCR7 favors antiplatelet lipids over an atherothrombotic lipidome and regulates thromboinflammation. Blood. 2022;139(11):1722–42. Ngamsri KC, Putri RA, Jans C, Schindler K, Fuhr A, Zhang Y, Gamper-Tsigaras J, Ehnert S, Konrad FM. CXCR4 and CXCR7 Inhibition Ameliorates the Formation of Platelet-Neutrophil Complexes and Neutrophil Extracellular Traps through Adora2b Signaling. Int J Mol Sci 2021, 22(24). Koch C, Engele J. Functions of the CXCL12 Receptor ACKR3/CXCR7-What Has Been Perceived and What Has Been Overlooked. Mol Pharmacol. 2020;98(5):577–85. Lounsbury N. Advances in CXCR7 Modulators. Pharmaceuticals (Basel) 2020, 13(2). Chagas MA, Babinski MA, Costa WS, Sampaio FJ. Stromal and acinar components of the transition zone in normal and hyperplastic human prostate. BJU Int. 2002;89(7):699–702. Gharaee-Kermani M, Kasina S, Moore BB, Thomas D, Mehra R, Macoska JA. CXC-type chemokines promote myofibroblast phenoconversion and prostatic fibrosis. PLoS ONE. 2012;7(11):e49278. Irani J, Levillain P, Goujon JM, Bon D, Dore B, Aubert J. Inflammation in benign prostatic hyperplasia: correlation with prostate specific antigen value. J Urol. 1997;157(4):1301–3. Motrich RD, Breser ML, Sanchez LR, Godoy GJ, Prinz I, Rivero VE. IL-17 is not essential for inflammation and chronic pelvic pain development in an experimental model of chronic prostatitis/chronic pelvic pain syndrome. Pain. 2016;157(3):585–97. Yi J, Pan J, Zhang S, Mao W, Wang J, Wang W, Yan Z. Improvement of chronic non-bacterial prostatitis by Jiedu Huoxue decoction through inhibiting TGF-beta/SMAD signaling pathway. Biomed Pharmacother. 2022;152:113193. Yi J, Pan J, Zhang S, Mao W, Wang J, Wang W, Yan Z. Jiedu Huoxue decoction improves chronic abacterial prostatitis/chronic pelvic pain syndrome through activating Wnt/GSKbeta/beta-catenin signaling pathway and alleviating apoptosis. Biomed Pharmacother. 2022;149:112830. Ho DR, Chang PJ, Lin WY, Huang YC, Lin JH, Huang KT, Chan WN, Chen CS. Beneficial Effects of Inflammatory Cytokine-Targeting Aptamers in an Animal Model of Chronic Prostatitis. Int J Mol Sci 2020, 21(11). Hu Y, Niu X, Wang G, Huang J, Liu M, Peng B. Chronic prostatitis/chronic pelvic pain syndrome impairs erectile function through increased endothelial dysfunction, oxidative stress, apoptosis, and corporal fibrosis in a rat model. Andrology. 2016;4(6):1209–16. Zhang S, Yue J, Ge Z, Xie Y, Zhang M, Jiang L. Activation of CXCR7 alleviates cardiac insufficiency after myocardial infarction by promoting angiogenesis and reducing apoptosis. Biomed Pharmacother. 2020;127:110168. Wei ST, Huang YC, Chiang JY, Lin CC, Lin YJ, Shyu WC, Chen HC, Hsieh CH. Gain of CXCR7 function with mesenchymal stem cell therapy ameliorates experimental arthritis via enhancing tissue regeneration and immunomodulation. Stem Cell Res Ther. 2021;12(1):314. Zhan CS, Chen J, Chen J, Zhang LG, Liu Y, Du HX, Wang H, Zheng MJ, Yu ZQ, Chen XG, et al. CaMK4-dependent phosphorylation of Akt/mTOR underlies Th17 excessive activation in experimental autoimmune prostatitis. FASEB J. 2020;34(10):14006–23. Sun X, Liu Y. Matrix Metalloproteinase-10 in Kidney Injury Repair and Disease. Int J Mol Sci 2022, 23(4). Ke B, Fan C, Yang L, Fang X. Matrix Metalloproteinases-7 and Kidney Fibrosis. Front Physiol. 2017;8:21. Lu SC. Regulation of glutathione synthesis. Mol Aspects Med. 2009;30(1–2):42–59. Pierre-Louis Odoom J, Freeberg MAT, Camus SV, Toft R, Szomju BB, Sanchez Rosado RM, Jackson PD, Allegood JC, Silvey S, Liu J, et al. Exhaled breath condensate identifies metabolic dysregulation in patients with radiation-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 2023;324(6):L863–9. Cho SJ, Moon JS, Nikahira K, Yun HS, Harris R, Hong KS, Huang H, Choi AMK, Stout-Delgado H. GLUT1-dependent glycolysis regulates exacerbation of fibrosis via AIM2 inflammasome activation. Thorax. 2020;75(3):227–36. Zeng H, Pan T, Zhan M, Hailiwu R, Liu B, Yang H, Li P. Suppression of PFKFB3-driven glycolysis restrains endothelial-to-mesenchymal transition and fibrotic response. Signal Transduct Target Ther. 2022;7(1):303. Cao H, Luo J, Zhang Y, Mao X, Wen P, Ding H, Xu J, Sun Q, He W, Dai C, et al. Tuberous sclerosis 1 (Tsc1) mediated mTORC1 activation promotes glycolysis in tubular epithelial cells in kidney fibrosis. Kidney Int. 2020;98(3):686–98. Srivastava SP, Li J, Kitada M, Fujita H, Yamada Y, Goodwin JE, Kanasaki K, Koya D. SIRT3 deficiency leads to induction of abnormal glycolysis in diabetic kidney with fibrosis. Cell Death Dis. 2018;9(10):997. Rao J, Wang H, Ni M, Wang Z, Wang Z, Wei S, Liu M, Wang P, Qiu J, Zhang L et al. FSTL1 promotes liver fibrosis by reprogramming macrophage function through modulating the intracellular function of PKM2. Gut 2022, 71(12):2539–2550. Mejias M, Gallego J, Naranjo-Suarez S, Ramirez M, Pell N, Manzano A, Suner C, Bartrons R, Mendez R, Fernandez M. CPEB4 Increases Expression of PFKFB3 to Induce Glycolysis and Activate Mouse and Human Hepatic Stellate Cells, Promoting Liver Fibrosis. Gastroenterology. 2020;159(1):273–88. Gao Y, Wei L, Wang C, Huang Y, Li W, Li T, Mo C, Qin H, Zhong X, Wang Y, et al. Chronic prostatitis alters the prostatic microenvironment and accelerates preneoplastic lesions in C57BL/6 mice. Biol Res. 2019;52(1):30. Okamoto K, Kurita M, Yamaguchi H, Numakura Y, Oka M. Effect of tadalafil on chronic pelvic pain and prostatic inflammation in a rat model of experimental autoimmune prostatitis. Prostate. 2018;78(10):707–13. He Y, Zeng HZ, Yu Y, Zhang JS, Duan X, Zeng XN, Gong FT, Liu Q, Yang B. Resveratrol improves prostate fibrosis during progression of urinary dysfunction in chronic prostatitis. Environ Toxicol Pharmacol. 2017;54:120–4. Zhang M, Liu Y, Chen J, Chen L, Zhang L, Chen X, Hao Z, Liang C. Targeting CXCL12/CXCR4 Signaling with AMD3100 Might Selectively Suppress CXCR4 + T-Cell Chemotaxis Leading to the Alleviation of Chronic Prostatitis. J Inflamm Res. 2022;15:2551–66. Gritsina G, Fong KW, Lu X, Lin Z, Xie W, Agarwal S, Lin D, Schiltz GE, Beltran H, Corey E et al. Chemokine receptor CXCR7 activates AURKA and promotes neuroendocrine prostate cancer growth. J Clin Invest 2023. Bai Y, Yang Y, Yan Y, Zhong J, Blee AM, Pan Y, Ma T, Karnes RJ, Jimenez R, Xu W, et al. RUNX2 overexpression and PTEN haploinsufficiency cooperate to promote CXCR7 expression and cellular trafficking, AKT hyperactivation and prostate tumorigenesis. Theranostics. 2019;9(12):3459–75. Li S, Fong KW, Gritsina G, Zhang A, Zhao JC, Kim J, Sharp A, Yuan W, Aversa C, Yang XJ, et al. Activation of MAPK Signaling by CXCR7 Leads to Enzalutamide Resistance in Prostate Cancer. Cancer Res. 2019;79(10):2580–92. Li R, Guan Z, Bi S, Wang F, He L, Niu X, You Y, Liu Y, Ding Y, Siwko S, et al. The proton-activated G protein-coupled receptor GPR4 regulates the development of osteoarthritis via modulating CXCL12/CXCR7 signaling. Cell Death Dis. 2022;13(2):152. Huebert RC, Shah VH. Sinusoidal endothelial cells direct traffic at the intersection of regeneration and fibrosis. Hepatology. 2014;60(2):754–6. Cao Z, Lis R, Ginsberg M, Chavez D, Shido K, Rabbany SY, Fong GH, Sakmar TP, Rafii S, Ding BS. Targeting of the pulmonary capillary vascular niche promotes lung alveolar repair and ameliorates fibrosis. Nat Med. 2016;22(2):154–62. Costello CM, McCullagh B, Howell K, Sands M, Belperio JA, Keane MP, Gaine S, McLoughlin P. A role for the CXCL12 receptor, CXCR7, in the pathogenesis of human pulmonary vascular disease. Eur Respir J. 2012;39(6):1415–24. Ghadge SK, Messner M, Seiringer H, Maurer T, Staggl S, Zeller T, Muller C, Bornigen D, Weninger WJ, Geyer SH et al. Smooth Muscle Specific Ablation of CXCL12 in Mice Downregulates CXCR7 Associated with Defective Coronary Arteries and Cardiac Hypertrophy. Int J Mol Sci 2021, 22(11). Additional Declarations No competing interests reported. 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-4186657\",\"acceptedTermsAndConditions\":true,\"allowDirectSubmit\":true,\"archivedVersions\":[],\"articleType\":\"Research Article\",\"associatedPublications\":[],\"authors\":[{\"id\":285608479,\"identity\":\"cfe4402a-bb40-49b7-8991-7b4ddd330536\",\"order_by\":0,\"name\":\"Yi zhang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Department of Urology, the First Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Yi\",\"middleName\":\"\",\"lastName\":\"zhang\",\"suffix\":\"\"},{\"id\":285608480,\"identity\":\"1c43989f-55bb-4456-8737-742c85c7e8fb\",\"order_by\":1,\"name\":\"Rui Feng\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Department of Urology, Shuguang Hospital, Shanghai University of Traditional Chinese Medicine, Shanghai 201203\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Rui\",\"middleName\":\"\",\"lastName\":\"Feng\",\"suffix\":\"\"},{\"id\":285608483,\"identity\":\"fd974130-af87-4b3d-95b2-2c9cffbe465a\",\"order_by\":2,\"name\":\"Weikang Wu\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Department of Urology, the First Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Weikang\",\"middleName\":\"\",\"lastName\":\"Wu\",\"suffix\":\"\"},{\"id\":285608485,\"identity\":\"29fb109d-e0f8-4b2f-8fc0-828427fc2aca\",\"order_by\":3,\"name\":\"Xianhong Liu\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Department of Urology, the First Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Xianhong\",\"middleName\":\"\",\"lastName\":\"Liu\",\"suffix\":\"\"},{\"id\":285608488,\"identity\":\"a927a61a-f02e-4435-9e5a-1da0932b7c09\",\"order_by\":4,\"name\":\"Cong Huang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Department of Urology, the First Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Cong\",\"middleName\":\"\",\"lastName\":\"Huang\",\"suffix\":\"\"},{\"id\":285608490,\"identity\":\"c560c914-3cb8-4171-b00e-26f3613ed057\",\"order_by\":5,\"name\":\"Xianguo Chen\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Department of Urology, the First Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Xianguo\",\"middleName\":\"\",\"lastName\":\"Chen\",\"suffix\":\"\"},{\"id\":285608492,\"identity\":\"08815eba-bfe1-4998-bf83-dc5ad70800c4\",\"order_by\":6,\"name\":\"Jing Chen\",\"email\":\"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA40lEQVRIiWNgGAWjYHACAwbGBgYGxmYg84OBjR1pWhhnFKQlE68FBJh5PhyCsXED/hnJGxh+7rDJY27nPfzaxuAAMwP74aMb8GmRuJFWwNh7Jq2YsZkvzTrH4A4fA09a2g281tzIMWBmbDuc2NjMY2acY/CMmUGCxwyvFnmIlv8QLRYGhxkbCGkxgGg5ANJi/JiBGC2GZ54B/dKWDLaFsccgLZmNkF/kjoNCrM0ucWP/GeMPP/7Y2PGzHz6G3/sCCew/wNY1MLBJgBhseJWDAP8BCC0PjMkPBFWPglEwCkbBiAQAEhdLMZAF5LsAAAAASUVORK5CYII=\",\"orcid\":\"\",\"institution\":\"Department of Urology, the First Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui\",\"correspondingAuthor\":true,\"prefix\":\"\",\"firstName\":\"Jing\",\"middleName\":\"\",\"lastName\":\"Chen\",\"suffix\":\"\"},{\"id\":285608493,\"identity\":\"dfc3941e-8189-4dd0-a43b-6fde2d0ef229\",\"order_by\":7,\"name\":\"Chaozhao Liang\",\"email\":\"\",\"orcid\":\"\",\"institution\":\"Department of Urology, the First Affiliated Hospital of Anhui Medical University, Anhui Medical University, Hefei, Anhui\",\"correspondingAuthor\":false,\"prefix\":\"\",\"firstName\":\"Chaozhao\",\"middleName\":\"\",\"lastName\":\"Liang\",\"suffix\":\"\"}],\"badges\":[],\"createdAt\":\"2024-03-29 08:31:41\",\"currentVersionCode\":1,\"declarations\":\"\",\"doi\":\"10.21203/rs.3.rs-4186657/v1\",\"doiUrl\":\"https://doi.org/10.21203/rs.3.rs-4186657/v1\",\"draftVersion\":[],\"editorialEvents\":[],\"editorialNote\":\"\",\"failedWorkflow\":false,\"files\":[{\"id\":54005912,\"identity\":\"a148c686-5b48-4a1b-84bd-f13e5f7b37f2\",\"added_by\":\"auto\",\"created_at\":\"2024-04-03 09:24:29\",\"extension\":\"png\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":3929395,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLegend not included with this version.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure1.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/959ba9ca9cb70dcfe310254c.png\"},{\"id\":54005300,\"identity\":\"a4f7c168-c855-455d-bf5a-e0269cb4e69f\",\"added_by\":\"auto\",\"created_at\":\"2024-04-03 09:16:28\",\"extension\":\"png\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":4366115,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLegend not included with this version.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure2.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/e0620b4b8932936237aa1383.png\"},{\"id\":54005304,\"identity\":\"de80d85a-9561-44f3-9937-25117ff2abff\",\"added_by\":\"auto\",\"created_at\":\"2024-04-03 09:16:29\",\"extension\":\"png\",\"order_by\":3,\"title\":\"Figure 3\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":1304006,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLegend not included with this version.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure3.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/d81d6b95324adab9a5b3b017.png\"},{\"id\":54005301,\"identity\":\"4a727f2f-5f78-4907-99e7-84a724e64eb5\",\"added_by\":\"auto\",\"created_at\":\"2024-04-03 09:16:29\",\"extension\":\"png\",\"order_by\":4,\"title\":\"Figure 4\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":658391,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLegend not included with this version.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure4.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/a33e6869b7a47c31af0f12c8.png\"},{\"id\":54005911,\"identity\":\"ace0284f-0614-4eb2-a428-233295458d95\",\"added_by\":\"auto\",\"created_at\":\"2024-04-03 09:24:29\",\"extension\":\"png\",\"order_by\":5,\"title\":\"Figure 5\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":3214199,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLegend not included with this version.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure5.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/39848262d5714c3d0f41689b.png\"},{\"id\":54005305,\"identity\":\"2a8d16c1-1587-46b3-9f4e-e894c1d3955b\",\"added_by\":\"auto\",\"created_at\":\"2024-04-03 09:16:29\",\"extension\":\"png\",\"order_by\":6,\"title\":\"Figure 6\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":5077344,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLegend not included with this version.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure6.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/da73cb314d2d2fc00f1284fd.png\"},{\"id\":54005307,\"identity\":\"0a69f521-56ef-4edd-806d-248894714016\",\"added_by\":\"auto\",\"created_at\":\"2024-04-03 09:16:29\",\"extension\":\"png\",\"order_by\":7,\"title\":\"Figure 7\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":616260,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eLegend not included with this version.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure7.png\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/28bfac88ec2c63c8b06e5657.png\"},{\"id\":54583099,\"identity\":\"072b1640-0ab9-4a19-bd70-8036810f8784\",\"added_by\":\"auto\",\"created_at\":\"2024-04-12 15:05:47\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":3147208,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-4186657/v1/d5dc2ddf-040f-4d91-b937-b3327daa57c5.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"The causative effect of CXCR7 on experimental autoimmune prostatitis injury and fibrosis\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eChronic prostatitis/Chronic pelvic pain syndrome (CP/CPPS) is one of the most common diseases of the urogenital system in young and middle-aged men, and is also listed as NIH Class III prostatitis, which is considered to be the most common type. The incidence is about 13.8% [1, 2].CP/CPPS is a disease that seriously affects the daily life of young and middle-aged men, and is characterized by pelvic perineal pain, obstructive or irritating lower urinary tract urination symptoms, and sexual dysfunction. The pathogenesis of CP/CPPS is complex, including bacterial infection, diet, immune cell infiltration, endocrine abnormalities [3] and Autonomic Nervous System Dysfunction [4]. However, the pathogenesis of CP/CPPS has not been clarified yet, so the therapeutic effect is not satisfactory. Therefore, it is necessary and necessary to find new therapeutic targets for CP/CPPS and explore new therapeutic methods.\\u003c/p\\u003e\\n\\u003cp\\u003eAlthough the role of CXCL12-CXCR4 signaling has been well studied, CXCR4 has high clinical value in hematopoietic stem cell and cancer biology, as well as in inflammatory diseases [5\\u0026ndash;8]. However, when chemokines participate in the inflammatory process, CXCR7, as a non-classical chemokine receptor, is up-regulated in many inflammatory diseases, such as rheumatoid arthritis [9], multiple sclerosis [10], atherosclerosis [11], and septic peritonitis [12]. However, unlike classical G protein-coupled receptors (GPCRS), CXCR7 does not signal through the G protein pathway, but instead the main ligands are the chemokines CXCL11 and CXCL12. Although CXCL12 has a greater affinity for CXCR7 than CXCR4, its receptor function is closely related to CXCR4. Depending on the cell type, CXCR4 either performs similar functional control in combination with CXCL12 or interacts with CXCR4 in quite complex and diverse ways [13, 14].\\u003c/p\\u003e\\n\\u003cp\\u003eDuring the progression of chronic inflammatory diseases, fibroblasts are activated to repair tissues abnormally, and tissue fibrosis is generally the end result of chronic inflammatory response, mainly the production of excessive collagen Abnormal activation of fibers and fibroblasts leads to organ dysfunction. In addition to smooth muscle cells, blood vessels and nerves, the prostate stroma is basically composed of collagen fibers, elastic fibers and undifferentiated fibroblasts [15]. Under the action of cytokines and chemokines (CXCL5, CXCL8, and CXCL12), intraprostatic myofibroblasts activate and secrete \\u0026alpha;-SMA at the site of inflammation [16], and may lead to permanent fibrosis in the subsequent progression of pathological processes. However, the effect of CXCR7 on prostatic inflammation and fibrosis in CP/CPPS remains unclear. In addition, CXCR7 agonists improved post-EAP mouse fibrosis by altering the spleen TH17/Treg cell ratio, and it is noteworthy that these conclusions are based on systematic inhibition of CXCR7 and CXCR4, making it an attractive target for therapeutic intervention.\\u003c/p\\u003e\\n\\u003cp\\u003eIn this study, we investigated the effects of CXCR7 on inflammation and the specific mechanisms that regulate fibrosis in EAP mouse models. Our results show that in mice treated with CXCR7 agonists, disease severity and inflammatory cell infiltration are reduced, and the progression of prostate fibrosis is alleviated through the TGF\\u0026beta;/SMAD pathway, reducing collagen accumulation.\\u0026nbsp;\\u003c/p\\u003e\"},{\"header\":\"Materials and Methods\",\"content\":\"\\u003cp\\u003eExperimental mice and EAP Induction\\u003c/p\\u003e \\u003cp\\u003eWe purchased male Sprague-Dawley rats from the Experimental Animal Center of Anhui Medical University for experiments. Prostate antigen preparation. Healthy male NOD/LtJ (NOD/ShiLtJ, 4 weeks old) mice. All mice, CNP Model Animal Research Center, Nanjing University (Nanjing).We domesticated them for a week before the experiment. All operations were performed under anesthesia. Chloral hydrate was used that the purpose is to reduce pain. Before the experiment began, all the mice underwent a week of acclimatization to the laboratory environment. All animals were kept in a dedicated pathogen free research animal facility at Anhui Medical University. In short, prostate homogenates were obtained from Sprague - Dawley rats and emulsified with superclear and equal amounts of full Freund adjuvant. The mice were immunized on day 0 and day 28 respectively according to the experimental plan. Then the experimental mice were injected with VUF11027(5mg/kg/ day, DMSO as a solvent) through the tail vein for two consecutive weeks. The mice were killed on day 50.\\u003c/p\\u003e \\u003cp\\u003eHematoxylin-Eosin Staining\\u003c/p\\u003e \\u003cp\\u003eProstate tissue was taken, fixed with 10% formalin solution, paraffin embedded, sliced, dewaxed to water, stained with hematoxylin dye, then dyed with eosin dye, and finally dehydrated and sealed. The degree of inflammation was quantified on a 4-point scale from 0 to 3 [\\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e], with the following criteria: 0, no inflammation; 1, mild inflammation, and clear monocytes around the blood vessels; 2, moderate inflammation, moderate perivascular monocytes surrounding; 3. Severe inflammation, bleeding, and a large number of monocytes in parenchyma with obvious infiltration around blood vessels.\\u003c/p\\u003e \\u003cdiv id=\\\"Sec2\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eIHC\\u003c/h2\\u003e \\u003cp\\u003eParaffin embedded tissue sections were dewaxed to water, heated at PH\\u0026thinsp;=\\u0026thinsp;6.0, 0.01M sodium citrate solution at 100\\u0026deg; for 15 minutes for antigen repair, and then put into 3% hydrogen peroxide solution to block endogenous peroxidase, and washed with PBS buffer three times for five minutes each time. Then it was sealed with 10% fetal bovine serum. The corresponding primary resistance was added and incubated at 4 degrees overnight. After washing with PBS for three times, the secondary antibody was added, incubated at room temperature for 2 hours, tested with freshly configured DAB chromogenic solution, and finally restained the nucleus with hematoxylin. The expression level was quantitatively analyzed using image J software\\u003c/p\\u003e \\u003cp\\u003eMasson staining\\u003c/p\\u003e \\u003cp\\u003eParaffin-embedded tissue sections were dewaxed to water, immersed in 2.5% potassium dichromate mordant overnight, and rinsed. The slices are then put into Weigert iron hematoxylin dye for 1min, then rinsed, and hematoxylin differentiation solution for 10sec. After rinsing with running water, the slices are placed into ponceau acid fuchsin dye solution for 10min. After rinsing with distilled water, the slices are placed into 1% phosphomolybdic acid solution for 3min. After rinsing and drying slightly, the slices are placed into 2.5% aniline blue solution for 1min and then rinsed and differentiated with 1% acetic acid. Dehydration with anhydrous ethanol three times, xylene transparent, neutral resin seal. image J software was used to quantitatively analyze the proportion of collagen fibers.\\u003c/p\\u003e \\u003cp\\u003eSirius Red staining\\u003c/p\\u003e \\u003cp\\u003eParaffin-embedded tissue sections were dewaxed to water, immersed in Sirius red staining for 10 minutes, dehydrated with anhydrous ethanol three times, xylene transparent section for 5 minutes, and sealed with neutral resin. image J software was used to quantitatively analyze the proportion of collagen fibers.\\u003c/p\\u003e \\u003cp\\u003eImmunofluorescence\\u003c/p\\u003e \\u003cp\\u003eThe paraffin sections were dewaxed to water, followed by antigen repair, sealing, adding primary antibody (CXCR7, ab72100, Abcam), and incubated in a wet box at 4\\u0026deg; overnight. After adding the secondary antibodies (IL17A, ab79056, Abcam), the cells were incubated for 90 minutes away from light, DAPI was re-dyed in the nucleus, and the cells were incubated at dark room temperature. The co-localization of CXCR7 and IL17A was confirmed by Olympus FV3000 confocal laser scanning microscope.\\u003c/p\\u003e \\u003cp\\u003eWestern Blotting Assays\\u003c/p\\u003e \\u003cp\\u003eThe protein extract was separated by 12.5% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred to polyvinylidene fluoride membrane. Incubate with 5% milk for 2 hours, then incubate with the corresponding primary antibody at 4\\u0026deg;C overnight. GAPDH was used as loading control. TBST washing membrane was incubated with the corresponding secondary antibody (anti-rabbit or anti-mouse IgG, 1:50,000, Elabscience) for 2 hours, and then washed with TBST. The membranes were washed, incubated with secondary antibodies for 2 h, and visualized using an EZ-ECL Kit (Biological Industries, Israel).\\u003c/p\\u003e \\u003cp\\u003eMouse Spleen Dissociation and Flow Cytometry\\u003c/p\\u003e \\u003cp\\u003eThe proportion of Th17 and Treg cells was measured by flow cytometry. The splenic single-cell suspension was prepared according to our previous method [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e]. Fitc-labeled CD4(BD Pharmingen, 557307,USA) and APC-labeled CD3e(BD Pharmingen, 553066,USA) were incubated with surface antibodies to APC-labeled CD25(BD Pharmingen, 553075,USA). After washing PBS again, Add 1 ml per tube containing phorbol 12myristate 13-acetate (PMA, MultiSciences, China), lonomycin (MultiSciences, China) 1640 medium (Gibco, USA) from monenin (MultiSciences, China) and Monenin (Multisciences, China) were incubated at 37℃ for 4 h and washed again. The cells were then sequentially immobilized and infiltrated, and incubated at 4 \\u0026deg; C for 60 min with intracellular antibodies, including PE-labeled IL-17A(BD Pharmingen, 559502, USA) and PE-labeled FoxP3(BD Pharmingen, 560401, USA). After washing again, analysis was performed with FACS Calibur flow cytometry (Beckman Coulter, USA). The resulting data is processed by FlowJo software (Tree Star, Ashland).\\u003c/p\\u003e \\u003cp\\u003emRNA Sequencing Experimental\\u003c/p\\u003e \\u003cp\\u003eTRIzol reagent was used to extract the total RNA of the sample. Smarter amplification kit was used to reverse transcribed the qualified total RNA and amplified the cDNA. The qualified cDNA was constructed by Tn5 transposition enzyme method: First, when the cDNA was randomly interrupted by transposase, the interrupted fragment was added with sequencing splices. Then, part of the sequencing splices were used as primers to amplify and enrich the library. After the amplified library was purified by magnetic beads and qualified by Agilent 2100 Bioanalyzer, Sequencing with Illumina Nova6000 other sequencers produced a double end number of 150bp. DESeq2 package was used to analyze the differentially expressed genes, which were consistent with P-value \\u0026amp;LT. Genes with a threshold of 0.05 and |LogFC2| \\u0026gt; 1 were defined as differentially expressed genes (DEGs).\\u003c/p\\u003e \\u003cp\\u003eUntargeted Metabolomics Analysis\\u003c/p\\u003e \\u003cp\\u003eWeigh the sample into a 1.5mL centrifuge tube, add two small steel balls, methanol-water (V:V\\u0026thinsp;=\\u0026thinsp;4:1, containing L-2-chlorophenylalanine, 4 \\u0026micro;g/mL); Put it in the refrigerator at -40℃ for 2 min, and then put it into the grinding machine (60 Hz, 2 min). Ultrasonic extraction in ice water bath for 10 min; Stand at -40℃ for 30 min; Centrifuge at low temperature for 10 min (12000 rpm, 4℃), and put the supernatant into a glass derived bottle; Use a centrifugal concentrator to dry the sample. Methoxyamine hydrochloride pyridine solution (15 mg/mL) was added to a glass derived vial and incubated at 37\\u0026deg;C for 60 min for oxime reaction. Remove samples after add 50 ul BSTFA derivatization reagent and 20 mu L n-hexane, add 10 kinds of internal standard 10 ul, 60 min at 70 ℃ reaction; After the samples were removed, they were placed at room temperature for 30 min for GC-MS analysis. The analytical instrument of the experiment was GMS-7890B /5977A (Aglient, USA).\\u003c/p\\u003e \\u003cp\\u003eWeigh the sample into a 1.5mL EP tube, add two small steel balls, and add methanol-water (V:V\\u0026thinsp;=\\u0026thinsp;4:1, containing L-2-chlorophenylalanine, 4 \\u0026micro;g/mL); After pre-cooling for 2 min in the \\u0026minus;\\u0026thinsp;40℃ refrigerator, put it into the grinder for grinding (60 Hz, 2 min); Ultrasonic extraction in ice water bath for 10 min and standing at -40℃ for 2 hours; Centrifuge for 10 min (12000 rpm, 4℃), the supernatant absorbed by syringe was filtered by 0.22 \\u0026micro;m organic phase pinhole filter, transferred to LC sample vial and stored at -80℃ until LC-MS analysis. Quality control sample (QC) is prepared by mixing the extract of all samples in equal volume. The analytical instrument for the experiment was ACQUITY UPLC I-Class system (Waters Corporation, USA).\\u003c/p\\u003e \\u003c/div\\u003e \\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e \\u003ch2\\u003eStatistical Analysis\\u003c/h2\\u003e \\u003cp\\u003eAll results were expressed as the mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;standard deviation (SD). GraphPad Prism version 8.0 software and (GraphPad Software, San Diego, CA).SPSS software version 26.0 (IBM Corp, Armonk, NY, USA) was used for statistical analysis. Two-tailed Student\\u0026rsquo;s t-tests or Wilcoxon rank sum test was applied for the comparison of two groups. Results were considered statistically significant if p-value\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05.\\u003c/p\\u003e \\u003c/div\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cp\\u003eCXCR7 agonists relieved inflammation in EAP mice\\u003c/p\\u003e \\u003cp\\u003eFollowing the procedure shown in Fig.\\u0026nbsp;1A, mice were successfully modeled using a mixture of Pags and full Fredringer adjuvant, followed by a 14-day caudal intravenous injection of CXCR7 agonist. Mechanical stimulation of the pelvic area showed that the response frequency of EAP mice was significantly higher than that of the normal group, while the response frequency after CXCR7 agonist was lower than that of the control group, and the response frequency was correlated with the applied force (Fig.\\u0026nbsp;1B). HE histological section (Fig.\\u0026nbsp;1C) showed that mice in EAP group had more inflammatory cell infiltration than mice in Control group, indicating that EAP mice were successfully induced. However, histopathological scores (Fig.\\u0026nbsp;1D) showed that inflammation was reduced after the administration of CXCR7 agonists compared to the control group who received tail vein DMSO. Chronic inflammation of prostate is closely related to cell necrosis and apoptosis, which has been confirmed in multiple experiments [\\u003cspan additionalcitationids=\\\"CR20 CR21\\\" citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e]. However, CXCR7 is involved in the process of apoptosis [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e], and it is obvious from immune tissue Caspase3 staining (Fig.\\u0026nbsp;1E) that the EAP group has more apoptosis than the Control group. The results of CXCR7 agonist and DMSO control group indicate that CXCR7 may also be involved in apoptosis in prostatitis.\\u003c/p\\u003e \\u003cp\\u003eCXCR7 aggravates prostatitis by affecting TH17/Treg cell imbalance\\u003c/p\\u003e \\u003cp\\u003eHE staining (Fig.\\u0026nbsp;1B) and immunohistochemical CD45 staining (Fig.\\u0026nbsp;2A) confirmed the increase of infiltrating inflammatory cells in the prostate of EAP mice. IL17A is highly expressed in the serum of prostatitis mice and has anti-inflammatory function [\\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e], which was also proved by ELISA experiments. CXCR7 can also reduce the expression of IL17A (Fig.\\u0026nbsp;2B). We speculated that CXCR7 has different expression of TH17 cells and Treg cells in prostatitis. Therefore, we conducted co-localization of CXCR7 and TH17 cells respectively (Fig.\\u0026nbsp;2C). Compared with the control group,CXCR7 agonist group showed that CXCR7\\u0026thinsp;+\\u0026thinsp;IL17A\\u0026thinsp;+\\u0026thinsp;cells could be reduced. Flow cytometry results showed that in the CXCR7 agonist group, the differentiation percentage of TH17 cells in spleen decreased (Fig.\\u0026nbsp;3A,C), while the differentiation percentage of Treg cells increased (Fig.\\u0026nbsp;3B,D). Taken together, these observations suggest that CXCR7 is involved in prostatic inflammation and that CXCR7 agonists restore EAP-induced Th17/Treg imbalances.\\u003c/p\\u003e \\u003cp\\u003eRNA-seq in EAP mice\\u003c/p\\u003e \\u003cp\\u003eIn order to investigate the mechanism of CXCR7 on the regulation of EAP mice, we divided the mice into EAP\\u0026thinsp;+\\u0026thinsp;VUF11207 group and EAP\\u0026thinsp;+\\u0026thinsp;DMSO group, respectively, and conducted rna sequencing analysis and off-target metabolic sequencing. By p\\u0026amp;lt; 0.05 and |logFC|\\u0026amp;gt; 1 is the cutoff value. Figure\\u0026nbsp;4A shows the heat map of the expression of differential genes in the two groups, while Fig.\\u0026nbsp;4B volcano map shows that 144 genes are up-regulated and 97 genes are down-regulated after CXCR7 agonist is used compared with the control group. Subsequently, Fig.\\u0026nbsp;4C and Fig.\\u0026nbsp;4D showed the pathway diagram of GO and KEGG enrichment analysis of differential genes, which can be seen as endopeptidase activity, serine\\u0026thinsp;\\u0026minus;\\u0026thinsp;type endopeptidase activity, serine\\u0026thinsp;\\u0026minus;\\u0026thinsp;type peptidase activity, serine hydrolase activity, etc. Matrix metalloproteinase is a classic hydrolase with endopeptidase activity, which can be secreted into extracellular matrix and deeply participate in the fibrosis process of organs [\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003euntargeted metabolomics in EAP mice\\u003c/p\\u003e \\u003cp\\u003eBecause we wanted to find out which metabolic processes and products regulated by CXCR7 in chronic prostatitis are involved in the occurrence and development of the disease, we conducted non-targeted metabolites detection on prostate tissues of EAP\\u0026thinsp;+\\u0026thinsp;VUF11207 group and EAP\\u0026thinsp;+\\u0026thinsp;DMSO group. In order to ensure more complete and comprehensive detection data, We used both LC-MS and GC-MS platforms for detection. In order to screen for differential metabolites, as shown in Fig.\\u0026nbsp;5A and Fig.\\u0026nbsp;5B, GC-MS and LC-MS products respectively, in which the red origin represents significantly up-regulated metabolites in the experimental group, the blue origin represents significantly down-regulated metabolites, and the gray dots represent insignificant metabolites. Correlation network (cornetwork) was used to calculate the correlation between differential metabolites and differential metabolite response intensity data based on pearson correlation analysis. pvalue \\u0026amp;lt was selected. 0.05,correlation \\u0026amp;gt; 0.95 relationship pairs are plotted in the network diagram (Fig.\\u0026nbsp;5C). Correlation analysis can help measure the degree of correlation between metabolites with significant differences, and further understand the interrelationship between metabolites in the process of biological state change. Correlation analysis used Pearson correlation coefficient to measure the degree of linear correlation between the two metabolites (Fig.\\u0026nbsp;5D).\\u003c/p\\u003e \\u003cp\\u003eThe TOP20 differential metabolic pathways were shown through the KEGG pathway mapper function, and the differential metabolites were shown according to the up-down-regulation information (Fig.\\u0026nbsp;5E). Notably, after CXCR7 agonist use, the Glycolysis/Gluconeogenesis pathway and Alanine, aspartate and glutamate metabolism pathway were identified in enrichment analysis. Glutamic acid is involved in the metabolic process of glutathione, which is considered to be a contributing factor to the mechanism of the pathological process of fibrosis in many pathological conditions [\\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e]. The involvement of glycolysis has been pointed out by many studies to be involved in the pathological process of fibrosis in various organs, such as lung [\\u003cspan citationid=\\\"CR29\\\" class=\\\"CitationRef\\\"\\u003e29\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR30\\\" class=\\\"CitationRef\\\"\\u003e30\\u003c/span\\u003e], heart [\\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e], kidney [\\u003cspan citationid=\\\"CR32\\\" class=\\\"CitationRef\\\"\\u003e32\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR33\\\" class=\\\"CitationRef\\\"\\u003e33\\u003c/span\\u003e], liver [\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e34\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e35\\u003c/span\\u003e], etc.\\u003c/p\\u003e \\u003cp\\u003eCXCR7 affects chronic prostatitis fibrosis\\u003c/p\\u003e \\u003cp\\u003eTo test whether CXCR7 affects prostatitis fibrosis, we administered CXCR7 agonists to EAP mice in a caudal vein. Firstly, Masson's (Fig.\\u0026nbsp;6A), Sirius Red (Fig.\\u0026nbsp;5B) and alfa-smooth muscle actin (α-SMA) (Fig.\\u0026nbsp;6C) staining were used to verify that EAP mice had fibrosis compared with normal mice. This was consistent with the results of other researchers [\\u003cspan additionalcitationids=\\\"CR37\\\" citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e36\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR38\\\" class=\\\"CitationRef\\\"\\u003e38\\u003c/span\\u003e], but it was also observed that the treatment of CXCR7 agonists significantly improved the fibrosis of prostate tissue compared with the treatment of DMSO controls. Our experimental results (Fig.\\u0026nbsp;6D-E) suggest that CXCR7 has an effect on chronic prostatitis fibrosis. These results suggest that CXCR7 is influencing prostatitis fibrotic process. To further identify CXCR7-mediated pathways of fibrotic processes. We found that VUF11207 improved the expression of TGFβ and decreased the phosphorylated protein level of SMAD2 (Fig.\\u0026nbsp;7A-C). At the protein level, the classic signature proteins of fibrosis, α-SMA (Fig.\\u0026nbsp;7A,D) and col1a1 (Fig.\\u0026nbsp;7A,E), were also significantly reduced after being treated with CXCR7 agonists. These results suggest an important role of CXCR7-mediated pathway in the progression of prostatitis fibrosis.\\u003c/p\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eOur previous findings determined that the CXCL12/ CXCR4 axis is involved in the pathogenesis and development of CNP by regulating the infiltration of CD4\\u003csup\\u003e+\\u003c/sup\\u003eT cells into the prostate gland and forming inflammatory nests [\\u003cspan citationid=\\\"CR39\\\" class=\\\"CitationRef\\\"\\u003e39\\u003c/span\\u003e]. However, the downstream ligands of CXCL12 are CXCR4 and CXCR7. Therefore, this study focused on the use of CXCR7 agonists. CXCR7 belongs to the superfamily of 7 transmembrane G-protein-coupled receptors, but cannot stimulate the typical G-protein-dependent pathway, but is involved in various pathophysiological functions. Recent studies have shown that the effects of CXCR7 on inflammation can be observed in a variety of diseases. Including cancer [\\u003cspan additionalcitationids=\\\"CR41\\\" citationid=\\\"CR40\\\" class=\\\"CitationRef\\\"\\u003e40\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR42\\\" class=\\\"CitationRef\\\"\\u003e42\\u003c/span\\u003e], bone and joint inflammation [\\u003cspan citationid=\\\"CR43\\\" class=\\\"CitationRef\\\"\\u003e43\\u003c/span\\u003e], atherosclerotic thrombosis [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e] and inflammatory demyelinating diseases [\\u003cspan citationid=\\\"CR10\\\" class=\\\"CitationRef\\\"\\u003e10\\u003c/span\\u003e].\\u003c/p\\u003e \\u003cp\\u003eFibrosis is a hallmark of various inflammatory processes. Many previous studies have fully demonstrated that CXCR7 is fully involved in the fibrosis process in the disease. The differential expression of SDF-1, CXCR7 and CXCR4 in hepatic sinusoidal endothelial cells leads to fibrotic transformation of vascular niches. selective CXCR7 activation in liver sinusoidal endothelial cells abrogated fibrogenesis[\\u003cspan citationid=\\\"CR44\\\" class=\\\"CitationRef\\\"\\u003e44\\u003c/span\\u003e]. Administration of CXCR7 agonists after lung injury can inhibit β-catenin-dependent Jag1 induction in injured PCEC[\\u003cspan citationid=\\\"CR45\\\" class=\\\"CitationRef\\\"\\u003e45\\u003c/span\\u003e]. CXCR7 mainly mediates regeneration, repair and proliferation of endothelial cells, and alveolar hypoxia can increase the expression of CXCR7 in lung endothelium [\\u003cspan citationid=\\\"CR46\\\" class=\\\"CitationRef\\\"\\u003e46\\u003c/span\\u003e]. In heart disease, CXCL12 is mainly expressed in vascular smooth muscle cells, and smooth muscle specific CXCL12 KO mutation leads to severe heart hypertrophy, and significant specific activation of pERK signal after CXCR7 agonist can alleviate the progression of myocardial hypertrophy in mice [\\u003cspan citationid=\\\"CR47\\\" class=\\\"CitationRef\\\"\\u003e47\\u003c/span\\u003e]. In conclusion, CXCR7 can influence fibrotic progression in inflammatory diseases.\\u003c/p\\u003e \\u003cp\\u003eIn this study, we found that CXCR7 in a mouse model of prostatitis affects the progression of chronic prostatitis and the development of TH17/Treg cell ratio in the model mice, and the fibrosis experiment further demonstrated that the functional level of CXCR7 affects the pathogenesis of chronic prostatitis fibrosis. In terms of mechanism, CXCL12/CXCR7 can regulate TGFβ-SMAD2/3 signaling pathway to promote the onset of chronic prostatitis and the progression of fibrosis. At present, the main drugs to treat chronic prostatitis are analgesics and proprietary Chinese medicine, and there is no effective treatment. Together, our results suggest that CXCR7 may be a promising therapeutic target for chronic prostatitis.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003eConflicts of interest disclosure\\u003c/p\\u003e\\n\\u003cp\\u003eNone.\\u003c/p\\u003e\\n\\u003cp\\u003eAvailability of Data and Materials\\u003c/p\\u003e\\n\\u003cp\\u003eNone.\\u003c/p\\u003e\\n\\u003cp\\u003eFunding support\\u003c/p\\u003e\\n\\u003cp\\u003eThe National Natural Science Foundation of China 82170787, 82370776, and 82300872.\\u003c/p\\u003e\\n\\u003cp\\u003eAuthor contribution\\u003c/p\\u003e\\n\\u003cp\\u003eLZ, XC and CJ: conception and design this study; ZY, FR and WWK: collection and assembly of data; WWK, HC, ZL,CJ: data Analysis and interpretation; ZY and FR: manuscript writing. Final Approval of Manuscript: All the authors.\\u003c/p\\u003e\\n\\u003cp\\u003eAcknowledgments\\u003c/p\\u003e\\n\\u003cp\\u003eWe wish to thank the Center for Scientific Research of the First Affiliated Hospital of Anhui Medical University for valuable help in our experiments.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\u003cli\\u003e\\u003cspan\\u003eKrieger JN, Nyberg L Jr., Nickel JC. NIH consensus definition and classification of prostatitis. JAMA. 1999;282(3):236\\u0026ndash;7.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBartoletti R, Cai T, Mondaini N, Dinelli N, Pinzi N, Pavone C, Gontero P, Gavazzi A, Giubilei G, Prezioso D, et al. Prevalence, incidence estimation, risk factors and characterization of chronic prostatitis/chronic pelvic pain syndrome in urological hospital outpatients in Italy: results of a multicenter case-control observational study. J Urol. 2007;178(6):2411\\u0026ndash;5. discussion 2415.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eStrauss AC, Dimitrakov JD. New treatments for chronic prostatitis/chronic pelvic pain syndrome. Nat Rev Urol. 2010;7(3):127\\u0026ndash;35.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHe H, Luo H, Qian B, Xu H, Zhang G, Zou X, Zou J. Autonomic Nervous System Dysfunction Is Related to Chronic Prostatitis/Chronic Pelvic Pain Syndrome. World J Mens Health 2023.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eDaniel SK, Seo YD, Pillarisetty VG. The CXCL12-CXCR4/CXCR7 axis as a mechanism of immune resistance in gastrointestinal malignancies. Semin Cancer Biol. 2020;65:176\\u0026ndash;88.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGupta N, Mohan CD, Shanmugam MK, Jung YY, Chinnathambi A, Alharbi SA, Ashrafizadeh M, Mahale M, Bender A, Kumar AP et al. CXCR4 expression is elevated in TNBC patient derived samples and Z-guggulsterone abrogates tumor progression by targeting CXCL12/CXCR4 signaling axis in mice model. Environ Res 2023:116335.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHeidegger I, Fotakis G, Offermann A, Goveia J, Daum S, Salcher S, Noureen A, Timmer-Bosscha H, Schafer G, Walenkamp A, et al. Comprehensive characterization of the prostate tumor microenvironment identifies CXCR4/CXCL12 crosstalk as a novel antiangiogenic therapeutic target in prostate cancer. Mol Cancer. 2022;21(1):132.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGallego C, Vetillard M, Calmette J, Roriz M, Marin-Esteban V, Evrard M, Aknin ML, Pionnier N, Lefrancois M, Mercier-Nome F, et al. CXCR4 signaling controls dendritic cell location and activation at steady state and in inflammation. Blood. 2021;137(20):2770\\u0026ndash;84.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWatanabe K, Penfold ME, Matsuda A, Ohyanagi N, Kaneko K, Miyabe Y, Matsumoto K, Schall TJ, Miyasaka N, Nanki T. Pathogenic role of CXCR7 in rheumatoid arthritis. Arthritis Rheum. 2010;62(11):3211\\u0026ndash;20.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003ePouzol L, Baumlin N, Sassi A, Tunis M, Marrie J, Vezzali E, Farine H, Mentzel U, Martinic MM. ACT-1004-1239, a first-in-class CXCR7 antagonist with both immunomodulatory and promyelinating effects for the treatment of inflammatory demyelinating diseases. FASEB J. 2021;35(3):e21431.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCebo M, Dittrich K, Fu X, Manke MC, Emschermann F, Rheinlaender J, von Eysmondt H, Ferreiros N, Sudman J, Witte A, et al. Platelet ACKR3/CXCR7 favors antiplatelet lipids over an atherothrombotic lipidome and regulates thromboinflammation. Blood. 2022;139(11):1722\\u0026ndash;42.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eNgamsri KC, Putri RA, Jans C, Schindler K, Fuhr A, Zhang Y, Gamper-Tsigaras J, Ehnert S, Konrad FM. CXCR4 and CXCR7 Inhibition Ameliorates the Formation of Platelet-Neutrophil Complexes and Neutrophil Extracellular Traps through Adora2b Signaling. \\u003cem\\u003eInt J Mol Sci\\u003c/em\\u003e 2021, 22(24).\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKoch C, Engele J. Functions of the CXCL12 Receptor ACKR3/CXCR7-What Has Been Perceived and What Has Been Overlooked. Mol Pharmacol. 2020;98(5):577\\u0026ndash;85.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLounsbury N. Advances in CXCR7 Modulators. \\u003cem\\u003ePharmaceuticals (Basel)\\u003c/em\\u003e 2020, 13(2).\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eChagas MA, Babinski MA, Costa WS, Sampaio FJ. Stromal and acinar components of the transition zone in normal and hyperplastic human prostate. BJU Int. 2002;89(7):699\\u0026ndash;702.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGharaee-Kermani M, Kasina S, Moore BB, Thomas D, Mehra R, Macoska JA. CXC-type chemokines promote myofibroblast phenoconversion and prostatic fibrosis. PLoS ONE. 2012;7(11):e49278.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eIrani J, Levillain P, Goujon JM, Bon D, Dore B, Aubert J. Inflammation in benign prostatic hyperplasia: correlation with prostate specific antigen value. J Urol. 1997;157(4):1301\\u0026ndash;3.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMotrich RD, Breser ML, Sanchez LR, Godoy GJ, Prinz I, Rivero VE. IL-17 is not essential for inflammation and chronic pelvic pain development in an experimental model of chronic prostatitis/chronic pelvic pain syndrome. Pain. 2016;157(3):585\\u0026ndash;97.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eYi J, Pan J, Zhang S, Mao W, Wang J, Wang W, Yan Z. Improvement of chronic non-bacterial prostatitis by Jiedu Huoxue decoction through inhibiting TGF-beta/SMAD signaling pathway. Biomed Pharmacother. 2022;152:113193.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eYi J, Pan J, Zhang S, Mao W, Wang J, Wang W, Yan Z. Jiedu Huoxue decoction improves chronic abacterial prostatitis/chronic pelvic pain syndrome through activating Wnt/GSKbeta/beta-catenin signaling pathway and alleviating apoptosis. Biomed Pharmacother. 2022;149:112830.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHo DR, Chang PJ, Lin WY, Huang YC, Lin JH, Huang KT, Chan WN, Chen CS. Beneficial Effects of Inflammatory Cytokine-Targeting Aptamers in an Animal Model of Chronic Prostatitis. Int J Mol Sci 2020, 21(11).\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHu Y, Niu X, Wang G, Huang J, Liu M, Peng B. Chronic prostatitis/chronic pelvic pain syndrome impairs erectile function through increased endothelial dysfunction, oxidative stress, apoptosis, and corporal fibrosis in a rat model. Andrology. 2016;4(6):1209\\u0026ndash;16.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZhang S, Yue J, Ge Z, Xie Y, Zhang M, Jiang L. Activation of CXCR7 alleviates cardiac insufficiency after myocardial infarction by promoting angiogenesis and reducing apoptosis. Biomed Pharmacother. 2020;127:110168.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eWei ST, Huang YC, Chiang JY, Lin CC, Lin YJ, Shyu WC, Chen HC, Hsieh CH. Gain of CXCR7 function with mesenchymal stem cell therapy ameliorates experimental arthritis via enhancing tissue regeneration and immunomodulation. Stem Cell Res Ther. 2021;12(1):314.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZhan CS, Chen J, Chen J, Zhang LG, Liu Y, Du HX, Wang H, Zheng MJ, Yu ZQ, Chen XG, et al. CaMK4-dependent phosphorylation of Akt/mTOR underlies Th17 excessive activation in experimental autoimmune prostatitis. FASEB J. 2020;34(10):14006\\u0026ndash;23.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSun X, Liu Y. Matrix Metalloproteinase-10 in Kidney Injury Repair and Disease. Int J Mol Sci 2022, 23(4).\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eKe B, Fan C, Yang L, Fang X. Matrix Metalloproteinases-7 and Kidney Fibrosis. Front Physiol. 2017;8:21.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLu SC. Regulation of glutathione synthesis. Mol Aspects Med. 2009;30(1\\u0026ndash;2):42\\u0026ndash;59.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003ePierre-Louis Odoom J, Freeberg MAT, Camus SV, Toft R, Szomju BB, Sanchez Rosado RM, Jackson PD, Allegood JC, Silvey S, Liu J, et al. Exhaled breath condensate identifies metabolic dysregulation in patients with radiation-induced lung injury. Am J Physiol Lung Cell Mol Physiol. 2023;324(6):L863\\u0026ndash;9.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCho SJ, Moon JS, Nikahira K, Yun HS, Harris R, Hong KS, Huang H, Choi AMK, Stout-Delgado H. GLUT1-dependent glycolysis regulates exacerbation of fibrosis via AIM2 inflammasome activation. Thorax. 2020;75(3):227\\u0026ndash;36.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZeng H, Pan T, Zhan M, Hailiwu R, Liu B, Yang H, Li P. Suppression of PFKFB3-driven glycolysis restrains endothelial-to-mesenchymal transition and fibrotic response. Signal Transduct Target Ther. 2022;7(1):303.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCao H, Luo J, Zhang Y, Mao X, Wen P, Ding H, Xu J, Sun Q, He W, Dai C, et al. Tuberous sclerosis 1 (Tsc1) mediated mTORC1 activation promotes glycolysis in tubular epithelial cells in kidney fibrosis. Kidney Int. 2020;98(3):686\\u0026ndash;98.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eSrivastava SP, Li J, Kitada M, Fujita H, Yamada Y, Goodwin JE, Kanasaki K, Koya D. SIRT3 deficiency leads to induction of abnormal glycolysis in diabetic kidney with fibrosis. Cell Death Dis. 2018;9(10):997.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eRao J, Wang H, Ni M, Wang Z, Wang Z, Wei S, Liu M, Wang P, Qiu J, Zhang L et al. FSTL1 promotes liver fibrosis by reprogramming macrophage function through modulating the intracellular function of PKM2. \\u003cem\\u003eGut\\u003c/em\\u003e 2022, 71(12):2539\\u0026ndash;2550.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eMejias M, Gallego J, Naranjo-Suarez S, Ramirez M, Pell N, Manzano A, Suner C, Bartrons R, Mendez R, Fernandez M. CPEB4 Increases Expression of PFKFB3 to Induce Glycolysis and Activate Mouse and Human Hepatic Stellate Cells, Promoting Liver Fibrosis. Gastroenterology. 2020;159(1):273\\u0026ndash;88.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGao Y, Wei L, Wang C, Huang Y, Li W, Li T, Mo C, Qin H, Zhong X, Wang Y, et al. Chronic prostatitis alters the prostatic microenvironment and accelerates preneoplastic lesions in C57BL/6 mice. Biol Res. 2019;52(1):30.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eOkamoto K, Kurita M, Yamaguchi H, Numakura Y, Oka M. Effect of tadalafil on chronic pelvic pain and prostatic inflammation in a rat model of experimental autoimmune prostatitis. Prostate. 2018;78(10):707\\u0026ndash;13.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHe Y, Zeng HZ, Yu Y, Zhang JS, Duan X, Zeng XN, Gong FT, Liu Q, Yang B. Resveratrol improves prostate fibrosis during progression of urinary dysfunction in chronic prostatitis. Environ Toxicol Pharmacol. 2017;54:120\\u0026ndash;4.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eZhang M, Liu Y, Chen J, Chen L, Zhang L, Chen X, Hao Z, Liang C. Targeting CXCL12/CXCR4 Signaling with AMD3100 Might Selectively Suppress CXCR4\\u0026thinsp;+\\u0026thinsp;T-Cell Chemotaxis Leading to the Alleviation of Chronic Prostatitis. J Inflamm Res. 2022;15:2551\\u0026ndash;66.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGritsina G, Fong KW, Lu X, Lin Z, Xie W, Agarwal S, Lin D, Schiltz GE, Beltran H, Corey E et al. Chemokine receptor CXCR7 activates AURKA and promotes neuroendocrine prostate cancer growth. J Clin Invest 2023.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eBai Y, Yang Y, Yan Y, Zhong J, Blee AM, Pan Y, Ma T, Karnes RJ, Jimenez R, Xu W, et al. RUNX2 overexpression and PTEN haploinsufficiency cooperate to promote CXCR7 expression and cellular trafficking, AKT hyperactivation and prostate tumorigenesis. Theranostics. 2019;9(12):3459\\u0026ndash;75.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLi S, Fong KW, Gritsina G, Zhang A, Zhao JC, Kim J, Sharp A, Yuan W, Aversa C, Yang XJ, et al. Activation of MAPK Signaling by CXCR7 Leads to Enzalutamide Resistance in Prostate Cancer. Cancer Res. 2019;79(10):2580\\u0026ndash;92.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eLi R, Guan Z, Bi S, Wang F, He L, Niu X, You Y, Liu Y, Ding Y, Siwko S, et al. The proton-activated G protein-coupled receptor GPR4 regulates the development of osteoarthritis via modulating CXCL12/CXCR7 signaling. Cell Death Dis. 2022;13(2):152.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eHuebert RC, Shah VH. Sinusoidal endothelial cells direct traffic at the intersection of regeneration and fibrosis. Hepatology. 2014;60(2):754\\u0026ndash;6.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCao Z, Lis R, Ginsberg M, Chavez D, Shido K, Rabbany SY, Fong GH, Sakmar TP, Rafii S, Ding BS. Targeting of the pulmonary capillary vascular niche promotes lung alveolar repair and ameliorates fibrosis. Nat Med. 2016;22(2):154\\u0026ndash;62.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eCostello CM, McCullagh B, Howell K, Sands M, Belperio JA, Keane MP, Gaine S, McLoughlin P. A role for the CXCL12 receptor, CXCR7, in the pathogenesis of human pulmonary vascular disease. Eur Respir J. 2012;39(6):1415\\u0026ndash;24.\\u003c/span\\u003e\\u003c/li\\u003e \\u003cli\\u003e\\u003cspan\\u003eGhadge SK, Messner M, Seiringer H, Maurer T, Staggl S, Zeller T, Muller C, Bornigen D, Weninger WJ, Geyer SH et al. Smooth Muscle Specific Ablation of CXCL12 in Mice Downregulates CXCR7 Associated with Defective Coronary Arteries and Cardiac Hypertrophy. Int J Mol Sci 2021, 22(11).\\u003c/span\\u003e\\u003c/li\\u003e\\u003c/ol\\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\":\"info@researchsquare.com\",\"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\":\"chronic prostatitis, CXCR7, inflammation, fibrosis\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-4186657/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-4186657/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003eChronic prostatitis and Pelvic Pain syndrome (CP/CPPS) is an autoimmune inflammatory disease characterized by pelvic or perineal pain and infiltration of inflammatory cells in the prostate. C-X-C chemokine receptor type 7 (CXCR7), also known as the atypical chemokine receptor 3 (ACKR3) receptor, are atypical chemokine receptors. Having been shown to play a key role in inflammatory processes, whether CXCR7 influences the role of autoimmune prostate and immune regulation and its mechanism of action are unclear. In this study, a mouse model of experimental autoimmune prostatitis was constructed by subcutaneous injection of antigen, and CXCR7 agonist was administered to investigate the effects of CXCR7 on the proportion of immune cells and fibrosis in CP/CPPS. Western blotting, immunohistochemical staining and Immunofluorescence, flow cytometry, and masson staining were used to study the possible regulatory mechanisms. CXCR7 agonists can significantly reduce pain and prostatic inflammation, and in vivo flow studies have shown that they affect the TH17/Treg cell ratio. To elucidate the potential mechanisms by which CXCR7 influences the pathogenesis of CNP, we conducted simultaneous RNA-seq and non-targeted metabolome sequencing. Our findings suggest that CXCR7 agonists alleviate fibrosis in autoimmune prostatitis by inhibiting the TGFβ/SMAD pathway. This study provides a valuable immunological basis for CNP to intervene CP/CPPS therapy with CXCR7 as the target.\\u003c/p\\u003e\",\"manuscriptTitle\":\"The causative effect of CXCR7 on experimental autoimmune prostatitis injury and fibrosis\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2024-04-03 09:16:24\",\"doi\":\"10.21203/rs.3.rs-4186657/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"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\":\"1b37f8c8-4b54-4224-8745-312347c15722\",\"owner\":[],\"postedDate\":\"April 3rd, 2024\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"posted\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2024-04-12T15:05:24+00:00\",\"versionOfRecord\":[],\"versionCreatedAt\":\"2024-04-03 09:16:24\",\"video\":\"\",\"vorDoi\":\"\",\"vorDoiUrl\":\"\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-4186657\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-4186657\",\"identity\":\"rs-4186657\",\"version\":[\"v1\"]},\"buildId\":\"qtupq5eGEP_6zYnWcrvyt\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}