The Causal Relationship between Chronic Obstructive Pulmonary Disease and Arterial Thrombotic Diseases: Role of Systemic Inflammation and NF- κB/COX-2 Pathway

The Causal Relationship between Chronic Obstructive Pulmonary Disease and Arterial Thrombotic Diseases: Role of Systemic Inflammation and NF- κB/COX-2 Pathway | 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 Article The Causal Relationship between Chronic Obstructive Pulmonary Disease and Arterial Thrombotic Diseases: Role of Systemic Inflammation and NF- κB/COX-2 Pathway You Wu, Houwen Zhang, Jialin Yu, Yu Liang, Wanru Cai This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-4384507/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 Background: Chronic Obstructive Pulmonary Disease (COPD) is a significant global health issue that often coexists with arterial thrombotic diseases. This study aims to investigate the causal relationship between COPD and these diseases, focusing on the role of systemic inflammation and the NF-κB/COX-2 pathway. Methods: The Two-Sample Mendelian Randomization (TSMR) approach was used to analyze the genetic correlation between COPD and the risks of ischemic stroke (IS) and acute myocardial infarction (AMI) using data from several large biobanks. Additionally, in vivo experiments with ApoE knockout mice and in vitro assays with primary mouse aorta endothelial cells were conducted to explore the role of the NF-κB/COX-2 pathway in COPD-related systemic inflammation. Results: The MR analysis revealed a significant association between COPD and increased risks of IS (OR: 1.152) and AMI (OR: 1.001). In vivo findings showed exacerbated pulmonary dysfunction and atherogenesis in mice with both COPD and high-fat diet (HFD), with notable histological changes in lung and aortic tissues. Inflammatory markers and lipid profiles were significantly altered in these models. In vitro studies demonstrated that COPD-induced systemic inflammation impaired endothelial cell function. These changes were mitigated by inhibiting the NF-κB/COX-2 pathway. Conclusions: This study provides strong evidence of a causal link between COPD and an elevated risk of arterial thrombotic diseases, mediated by systemic inflammation and the NF-κB/COX-2 pathway. These findings highlight the importance of addressing arteriosclerosis and thrombosis formation risks in COPD management and suggest that the NF-κB/COX-2 pathway could be a potential therapeutic target for reducing comorbidity in COPD patients. Biological sciences/Genetics Biological sciences/Immunology Biological sciences/Neuroscience Health sciences/Diseases/Cardiovascular diseases Health sciences/Diseases/Neurological disorders Health sciences/Diseases/Respiratory tract diseases Chronic obstructive pulmonary disease inflammation nuclear factor-κB cyclooxygenase-2 Mendelian Randomization Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 6 Introduction Chronic obstructive pulmonary disease (COPD) is a heterogeneous lung condition. [ 1 ] It is a leading cause of death worldwide and poses a significant threat to public health. COPD often coexists with various diseases, particularly cardiovascular and cerebrovascular diseases. [ 2 ] Numerous studies have suggested that COPD independently increases the risk of cardiovascular diseases. [ 3 – 5 ] Therefore, it is crucial to investigate the complex relationships and underlying mechanisms between COPD and cardiovascular disorders to enhance prevention, treatment, and overall management strategies. COPD involves the activation of multiple inflammatory cells and factors, which contribute to the inflammatory response. [ 6 , 7 ] COPD patients experience persistent inflammation not only in the lungs but also throughout their entire system. [ 8 , 9 ] Some research has shown that this systemic inflammation increases the risk of various health issues, such as diabetes, stroke, cardiovascular diseases, lung cancer, and pneumonia, by 2–4 times in COPD patients. [ 10 ] Therefore, systemic inflammatory responses are considered significant contributors to the development and progression of these comorbidities in individuals with COPD. [ 11 ] It is well-established that arteriosclerosis is a primary factor in the onset of myocardial infarction and stroke. A large body of compelling experimental and clinical data now indicates that inflammation participates fundamentally in atherogenesis (AS) and in the pathophysiology of ischaemic events. [ 12 ] Research shows that NF-κB is one of the initiating mechanisms for vascular endothelial cell injury. [ 13 ] Activated NF-κB can induce the continuous expression of cytokines, adhesion molecules, and enzymes related to the amplification of the inflammatory cascade. [ 14 , 15 ] Adhesion molecules such as monocyte chemoattractant protein-1 (MCP-1), intercellular cell adhesion molecule-1 (ICAM-1), vascular cellular adhesion molecule-1 (VCAM-1), and the selectin family are also expressed. [ 16 , 17 ] NF-κB can increase the expression of inflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-8 (IL-8). Conversely, these inflammatory mediators can also promote the activation of NF-κB, thereby amplifying the inflammatory response. [ 18 ] Upon activation, NF-κB also induces the enhanced expression of cyclooxygenase-2 (COX-2). Once activated, COX-2 can both reduce the bioavailability of nitric oxide (NO) and promote the accumulation of reactive oxygen species, intensifying oxidative stress. [ 18 , 19 ] It can also catalyze the production of prostaglandin E2 (PGE2), activating inflammatory cells at the site of inflammation, inducing the release of chemokines, recruiting inflammatory cells, and inducing the production of multiple inflammatory factors, thereby maintaining and amplifying the inflammatory cascade. [ 20 – 22 ] These factors increase vascular permeability and uptake of oxidized low-density lipoprotein cholesterol (OxLDL) [ 23 ] promoting plaque formation and participating in the onset and development of AS. Studies have shown that by inhibiting the activity of NF-κB and suppressing the expression of COX-2 messenger RNA (mRNA), the progression of inflammation can be inhibited, providing a protective effect on endothelial cells. [ 24 , 25 ] Mendelian randomization (MR) analysis has emerged as a potent method for identifying causal relationships between risk factors and diseases, using genetic variants as instrumental variables (IVs). Building on this, the current study utilizes two-sample Mendelian randomization (TSMR) analysis to ascertain the potential causal link between COPD and the risk of developing cardiovascular and cerebrovascular diseases. Additionally, through both in vivo and in vitro experiments, we confirmed that systemic inflammation induced by COPD promotes atherogenesis lesions by activating the NF-κB/COX-2 pathway. [ 26 ] Methods Mendelian Randomization Analysis This study employed the TSMR approach, considering COPD as the exposure factor and both stroke and myocardial infarction as outcome variables to establish causal links. Data related to COPD, ischemic stroke (IS) [ 27 ] , and acute myocardial infarction (AMI) were respectively sourced from FinnGen Biobank, MEGASTROKE, and the MRC-IEU organizations (Table 1 ). Table 1 Characteristics of data sources and strength of IVs used in the Mendelian randomization study Exposures/Outcomes GWAS ID Consortium Ethnicity Sample Sizes Number of SNPs Year COPD finn-b-J10_COPD FinnGen Biobank European 193,638 16,380,382 2021 Ischemic stroke ebi-a-GCST006907 MEGASTROKE European 150,765 8,418,349 2018 Acute myocardial infarction ukb-b−3469 MRC-IEU European 463,010 9,851,867 2018 GWAS:Genome-Wide Association Studies,COPD:Chronic obstructive pulmonary disease,SNPs: single nucleotide polymorphisms, NA: Not Available. To conduct the analysis, we compiled single nucleotide polymorphisms (SNPs) associated with COPD at a significant level of P < 5 × 10 − 6 . In order to ensure the independence of these SNPs, we established a linkage disequilibrium threshold (r 2 ) of 0.01 and a genetic distance of 5000 kb for the selection process. We identified the phenotypes associated with the remaining SNPs using the Human Genotype-Phenotype Association Database ( www.phenoscanner.medschl.cam.ac.uk ), [ 28 ] with a special emphasis on excluding SNPs whose corresponding phenotypes were significantly associated with the outcomes. The datasets for both the exposure and outcome variables were combined, while palindromic SNPs were removed to ensure the integrity of the analysis. In the meanwhile, to mitigate interference from reverse causality, reverse MR analysis was employed using SNPs related to IS and AMI. Mice 8-week-old male SPF grade ApoE knockout mice (ApoE−/−) were purchased from the Experimental Animal Center of Hangzhou Medical College in Zhejiang, China (Licence Number: SYXK(Zhe)2019-0011) and housed in a specific pathogen-free (SPF) facility in a controlled environment. All experimental protocols were approved by the Zhejiang Provincial Experimental Animal Center Animal Welfare Ethics Committee (Ethics Number: ZJCLA-IACUC-20030066). All methods were carried out in accordance with relevant guidelines and regulations. All animal experiments were taken place in SPF Animal Laboratory at Zhejiang Chinese Medical University. The mice were randomly divided into the following groups: Control group: Mice were given a normal diet, and equivalent doses of saline were instilled during the PPE and LPS induction for the COPD model. COPD group: Mice were instilled with 1.2 IU of PPE via the airway once a week for a total of 4 times. Two weeks after the final PPE instillation, 200 µg of LPS was instilled once a week, for a total of 2 times. HFD group: To establish the AS model, mice were fed a HFD (comprising 3% cholesterol, 0.5% sodium cholate, 0.2% propylthiouracil, 5% sugar, 10% lard, and 81.3% base diet) for 7 weeks. HFD + COPD group: Alongside initiating HFD, COPD interventions were given as well. Mice received 1.2 IU of PPE instilled via the airway once a week for a total of 4 times. One week after the last PPE instillation, 200 µg of LPS was instilled once a week for a total of 2 times. HFD + COPD + BAY11-7082 group: While establishing the combined COPD and AS model, mice were given intraperitoneal injections of the NF-κB inhibitor BAY11-7082 (HY-13453, MedChemExpress) (5 mg/kg) on the same day as the PPE instillation. Subsequently, injections were given every other day until the end of the experiment. HFD + COPD + NS-398 group: While establishing the combined COPD and AS model, mice were given intraperitoneal injections of the COX-2 inhibitor NS-398 (HY-13913, MedChemExpress) (5 mg/kg) on the same day as the PPE instillation. Subsequently, injections were given every other day until the end of the experiment. Cells Primary mouse aorta endothelial cells were used for this study. The aortic endothelial cells were then purified using anti-CD31 coupled magnetic beads. [ 29 ] ( Fig. 5 A-B ) Simultaneously, serum was extracted from both the COPD group of mice and the control group of mice. The cells were randomly divided into the following groups: Control group: cells received no special treatment. NS group: cells received serum from the control group mice. CS group: cells received serum from COPD mice. CS + BAY11-7082 group: cells received BAY11-7082 while receiving serum from COPD mice. CS + BAY11-7082 + oe-NC group: cells received serum from COPD mice, BAY11-7082, and transfected empty plasmid simultaneously. CS + BAY11-7082 + oe-COX-2 group: cells received serum from COPD mice, BAY11-7082, and transfected COX-2 expressing plasmid (purchased from Santa Cruz Biotechnology) simultaneously. Pulmonary Function Test Mice were anaesthetized by intraperitoneal injection using a 3% pentobarbital sodium solution. After the tracheal cannula was connected to the airway of the plethysmograph (PFT, DSI), the measurements of functional residual capacity (FRC), resistance index (RI), dynamic lung compliance (Cdyn), and minute ventilation (MV) were taken. At the end of these experiments, all mice were euthanized by CO 2 asphyxiation. Mice Lipid Profile The following mouse kits were used: Total Cholesterol (TC), Triglyceride (TG), High-Density Lipoprotein Cholesterol (HDL-C), and Low-Density Lipoprotein Cholesterol (LDL-C) (A111-1-1, A110-1-1, A112-1-1 and A113-1-1, Nanjing Jiancheng Bioengineering Institute). These serum samples then underwent testing on a fully-automated biochemistry analyser (COBAS INTEGRA 800, Roche, Switzerland). Mice Tissue The lung tissue and aortic roots of the different groups of mice were evaluated using Hematoxylin and Eosin (H&E) (C0105S, Beyotime) staining to assess the pathological changes in lung tissues and the size of arteriosclerotic plaques. Additionally, Oil Red O (G1015-100ML, Servicebio) staining was employed to evaluate lipid accumulation in the aorta. Cells Functions Cells were seeded in a 96-well plate, treated with CCK-8 (CA1210, Solarbio) solution, and absorbance at 450 nm was measured to determine viability. Apoptosis was evaluated using flow cytometry (Attune™ NxT, Thermo Fisher Scientific), where cells were stained with Annexin V-FITC (C1062S, Beyotime) and Propidium Iodide (PI), and apoptotic cells were detected based on fluorescence microscope (M205 FCA, Leica). Cells were seeded in Matrix-Gel™ chambers, cultured, and stained with crystal violet for migration assessment. Tubule formation potential was investigated by starving cells, seeding them in Matrigel-coated wells, and observing the formation of tube networks. RT-qPCR Total RNA was extracted from the cells using TRIzol reagent. cDNA was then synthesized using PrimeScript reverse transcriptase following the manufacturer's protocol. The amplified products were COX-2 forward primer: 5'-AGGACTCTGCTCACGAAGGA-3', COX-2 reverse primer: 5'- TGACATGATTGGAACAGCA-3', GAPDH forward primer: 5'-ACCCTTAAGAGGATGCTGC-3', and GAPDH reverse primer: 5'- CCCAATACGGCCAAATCCGT-3'. The gene expression levels were quantified using the 2^-ΔΔCQ method and normalized to the internal reference gene GAPDH. ELISA The relevant inflammatory factors in the mice aortic tissues, serum, and cells from each group were detected using enzyme-linked immunosorbent assay (ELISA) kits for IL-6, IL-1β, TNF-α, PGE2, MCP-1, VCAM-1, and ICAM-1(SEKM-0007, SEKM-0002, SEKM-0034, SEKM-0173, SEKM-0108, SEKM-0037, SEKM-0132). Analyses were performed according to the kit instructions. Western Blot Proteins were extracted from the aortic tissues of various mouse groups and from the cells of each group using RIPA lysis buffer (D3910201, Sigma) supplemented with 1% Phenylmethanesulfonyl fluoride (PMSF). Proteins were then separated by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), with an equal amount of protein (30µg) loaded per lane, and subsequently transferred onto PVDF membranes (IPVH00010, Millipore). These membranes were incubated overnight at 4°C with antibodies diluted at 1:3000 for p-p65, p65, COX-2, caspase 3, Bcl-2, Bax, and GAPDH (3033, 8242, 4842, 9662, 3498, 2772 and 5174, CST). This was followed by a 1-hour incubation with Horseradish Peroxidase (HRP) conjugated secondary antibodies. Densitometric analysis was conducted using ImageJ software. 4. Statistical Analysis We utilized the software packages "TwoSampleMR" and "MRPRESSO" with R version 4.3.1 to conduct MR analysis. We utilized several methods including inverse variance weighting (IVW), [ 30 , 31 ] MR-Egger, [ 32 , 33 ] Simple mode, weighted mode and weighted median method. [ 34 ] The IVW method was used as the primary analysis. We assessed heterogeneity by employing Cochran’s Q method, considering heterogeneity to be significant at p < 0.05. [ 35 ] We utilized the MR-Egger intercept test [ 33 ] and the MR-PRESSO global test to identify and remove any potential outliers with horizontal pleiotropic effects that might have significantly influenced the estimation results. [ 36 ] This study utilized SPSS 22.0 software for the in vivo and in vitro experiments statistical analysis. Quantitative data were assessed for normality and presented as the mean ± standard deviation (x̅ ± SD). The T test was employed to analyse differences between groups, and statistical significance was determined when P < 0.05. Graphs were generated using GraphPad Prism 8.0 (GraphPad, USA). Results MR Analysis In our MR analysis investigating the impact of COPD on cardiovascular and cerebrovascular diseases, a meticulous selection of 19 SNPs associated with the influence of COPD on IS, and 7 SNPs associated with AMI was conducted ( Supplementary Tables ). Using the random-effects IVW method, a significant positive association was observed between COPD and the risk of IS (OR: 1.152, 95% CI: 1.022-1.300, P = 0.021) and AMI (OR: 1.001, 95% CI: 1.001–1.002, P = 0.029) (Fig. 1 ). These findings were consistently supported across various methodologies. Sensitivity analyses, encompassing MR-Egger intercepts and Cochran Q tests, did not reveal significant directional pleiotropy, heterogeneity, or notable outliers, further substantiating the robustness of our results ( Supplementary Tables ). Moreover, no single SNP was found to disproportionately influence the overall causal estimates, as confirmed by the leave-one-out analysis ( Supplementary Fig. 1 ), thereby underscoring the stability and reliability of our study outcomes. For the reverse MR, no significant effect of IS and AMI on COPD was observed ( Supplementary Tables ). Enhanced Pulmonary Dysfunction and Atherogenesis in COPD and HFD Mice Our investigation revealed significant functional alterations in the pulmonary system in different mouse models. In both the COPD and HFD groups, there was a noticeable increase in FRC and RI, along with a significant decrease in Dynamic Lung Cdyn and MV, compared to the control group (Fig. 2 A-D). The decline in lung function was even more prominent in the HFD + COPD group. Histological Analysis of Lung and Aortic Tissues In the lung tissues, while the control group displayed intact alveolar structures, the COPD and HFD groups showed notable reductions in alveolar number accompanied by substantial macrophage infiltration (Fig. 2 I). This degeneration was even more pronounced in the HFD + COPD group. Interestingly, treatment with NF-κB and COX-2 inhibitors, particularly in the HFD + COPD + BAY11-7082 and HFD + COPD + NS-398 groups, led to a significant recovery in alveolar count and structural integrity. In the aortic tissues, changes were equally remarkable. The control group showed minimal atherosclerotic changes, whereas the HFD group exhibited a noticeable increase in atheromatous plaque formation. This atherosclerotic progression was even more extensive in the HFD + COPD group, with larger plaque formations (Fig. 3 A and B ). However, in the groups treated with NF-κB and COX-2 inhibitors, there was a notable reduction in the size and complexity of atherosclerotic plaques. Lipid Profile Alterations and Inflammatory Marker Dynamics Lipid profile analysis revealed a significant dysregulation in the HFD group, with elevated levels of TC, TG and LDL-C, coupled with a decrease in HDL-C (Fig. 2 E-H). This dysregulation was exacerbated in the HFD + COPD group, suggesting that COPD may intensify lipid abnormalities in atherosclerosis. Inflammatory markers such as IL-6, IL-1β, TNF-α, PGE2, MCP-1, VCAM-1, and ICAM-1 displayed a significant increase in the COPD and HFD groups (Fig. 4 ). This increase was even more pronounced in the HFD + COPD group, indicating an enhanced inflammatory response due to the combination of COPD and atherosclerosis. Apoptosis and NF-κB/COX-2 Pathway Activation in Aortic Tissues Western Blot analysis of aortic tissues from the COPD and HFD groups revealed an upregulation of apoptosis-related proteins Cleaved-caspase 3 and Bax, alongside a downregulation of Bcl-2, with the most significant changes observed in the HFD + COPD group (Fig. 3 C and D ). The activation of the NF-κB/COX-2 pathway was also apparent in these groups, as demonstrated by elevated expressions of p-65, p-p65 and COX-2 (Fig. 3 E and F ). These molecular changes were significantly altered upon treatment with NF-κB and COX-2 inhibitors. Endothelial Cell Function Under COPD-Induced Systemic Inflammation In vitro experiments utilizing primary aortic endothelial cells exposed to serum from COPD mice (CS group) revealed a reduction in cell viability and an increase in apoptosis rates compared to cells exposed to control serum (NS group). The application of BAY11-7082, an NF-κB inhibitor, markedly improved cell viability and reduced apoptosis rates (Fig. 5 C-F). However, overexpression of COX-2 in these cells negated the protective effects of BAY11-7082. Migration, Tubule Formation, and Inflammatory Responses in Endothelial Cells Cell migration and tubule formation capabilities were assessed to understand the effects of COPD-induced systemic inflammation on endothelial cell function. A significant reduction in cell migration (Fig. 6 A) and tubule formation (Fig. 6 B) was observed in the CS group, which was reversed upon treatment with BAY11-7082. However, this reversal was diminished with COX-2 overexpression. Additionally, inflammatory markers in these cells showed an increase in the CS group, which was reduced following treatment with BAY11-7082 (Fig. 6 C). Yet, this reduction was counteracted by COX-2 overexpression. NF-κB/COX-2 Pathway Influence in Endothelial Cells Detailed analysis through immunofluorescence (Fig. 6 D) and Western Blot (Fig. 6 E and F ) techniques revealed that exposure to COPD-induced serum led to enhanced nuclear translocation of p65 and increased expressions of p-p65 and COX-2 in endothelial cells. These molecular changes were attenuated by the NF-κB inhibitor BAY11-7082 but were reinstated upon COX-2 overexpression. Discussion In this study, we utilized a TSMR approach to investigate the causal relationship between COPD and the increased risks of IS and AMI. Our analysis, which involved carefully selecting SNPs from large-scale biobanks, revealed a significant genetic correlation between COPD and these cardiovascular conditions. Additionally, we conducted complementary in vivo and in vitro experiments to further elucidate the role of systemic inflammation, specifically through the NF-κB/COX-2 pathway, in mediating this relationship. Multiple studies have consistently demonstrated elevated levels of proinflammatory cytokines, including serum C-reactive protein, plasma fibrinogen, serum amyloid A, TNF-α, IL-6, IL-8, COX-2, etc. in patients with COPD. [ 37 – 41 ] This systemic inflammation is further exacerbated during acute exacerbations of COPD. [ 29 , 41 – 43 ] In our COPD model, we also observed a corresponding increase in these inflammatory markers. However, the exact mechanism underlying this phenomenon remains unclear, possibly attributed to the leakage of various proinflammatory markers from the lungs into the bloodstream. These findings provide evidence for the presence of a systemic inflammatory response in individuals with COPD. [ 44 , 45 ] The NF-κB/COX-2 inflammatory pathway plays a crucial role in regulating AS. Gareus et al. [ 46 ] conducted research demonstrating that the depletion of key signaling molecules in the NF-κB pathway, namely nuclear factor κB kinase γ (IKKγ) or nuclear factor κB inhibitor α (IkappaBalpha, IκBα), significantly reduced the formation of atherosclerotic plaques in ApoE−/− mice. Additionally, circulating microRNAs (miRNAs) have been found to reduce inflammation and lesions in ApoE−/− atherosclerotic mice by inhibiting the activation of NF-κB in vascular endothelial cells. [ 47 ] This research suggests that the key vault protein can suppress the NF-κB signal-mediated inflammatory pathway, thereby inhibiting obesity and the development of AS. [ 48 ] Moreover, methyltransferase-like 14 (Mettl14) has been implicated in the inflammatory response of macrophages in AS through the NF-κB/IL-6 signaling pathway. [ 49 ] Studies have demonstrated that inhibiting NF-κB activity to suppress the expression of COX-2 mRNA can reduce oxidative stress, inhibit inflammation, and protect endothelial cells. [ 50 , 51 ] Similar results were obtained when separately inhibiting NF-κB and COX-2. Although the NF-κB/COX-2 signaling pathway has not been validated in the study of COPD combined with AS, it has been shown to have a significant impact on other diseases, particularly those related to inflammation. Research has demonstrated that inflammatory factors stimulate the translocation of NF-κB in human bronchial smooth muscle cells, and the activation of NF-κB in the airway epithelium leads to a significant increase in inflammatory factors. [ 52 ] The NF-κB signaling pathway plays a crucial role in regulating inflammation through COX-2 transcription. For example, Seyyed and colleagues' research indicates that NF-κB-induced COX-2 is an important factor in causing allergic asthma lung inflammation. [ 53 ] In a mouse model of LPS-induced liver injury, lycopene has been shown to alleviate liver function damage by inhibiting NF-κB/COX-2 signal transmission and reducing the expression of inflammatory factors in mouse serum. [ 54 ] In a rat model of pharyngitis, the NF-κB/COX-2 signaling pathway was significantly activated, and inhibiting the activity of this pathway alleviated pharyngitis-related throat symptoms. [ 55 ] HE staining revealed increased inflammation indicators in arterial tissue and elevated inflammatory factors in serum of the HFD mouse group, accompanied by a decrease in the number of alveoli and minor macrophage infiltration within the alveoli. This may be related to high-fat diet and further experiments are needed to verify. [ 56 , 57 ] In two intervention experiments and treatments overexpressing COX-2, we noted a clear upstream-downstream relationship between NF-κB and COX-2 but have yet to investigate how to further enhance the expression of inflammatory factors subsequent to COX-2 expression. One potential mechanism could be that COX-2 synthesized PGE2, after binding with its receptors EP2 and EP4, activates adenylyl cyclase in cells causing increased intracellular cAMP levels. [ 58 ] Heightened cAMP levels further stimulate protein kinase A (PKA), which can phosphorylate and activate the transcription factor CREB (cAMP response element binding protein), promoting gene transcription of inflammation factors such as IL-6 and TNF-α. Additionally, PGE2 can promote the gene transcription of inflammatory factors such as IL-6 and TNF-α by activating the MAPK signaling pathway (including ERK, JNK and p38 kinases). [ 59 , 60 ] The NF-κB/COX-2 inflammatory pathway presents an opportunity for targeting the prevention of COPD in combination with AS and the occurrence of malignant ischemic events. However, the safety and effectiveness of potential drugs such as Murraya paniculata, [ 61 ] Berberine Targets, [ 62 ] Eugenol, [ 63 ] etc. still require further verification through clinical practice. Conclusions The convergence of genetic, clinical, and molecular data in our study provides strong evidence supporting the integration of arterial thrombotic disease risk management in the treatment and overall care strategies for patients with COPD. Furthermore, our findings suggest that targeting the NF-κB/COX-2 pathway could be a potential therapeutic approach to mitigate the increased arteriosclerosis and thrombosis formation risks associated with COPD, opening up new avenues for treatment strategies. In conclusion, this study improves our understanding of COPD as a systemic disease with significant cardiovascular implications. It highlights the importance of a comprehensive approach to managing COPD, addressing both pulmonary and cardiovascular aspects, in order to achieve better patient outcomes. Declarations Ethics approval Data for the Mendelian andomization analysis was sourced from publicly available databases, negating the need for ethical approval or patient consent for this analysis. All animal experiments were taken place in SPF Animal Laboratory at Zhejiang Chinese Medical University and were in compliance with the ethical requirements of the Zhejiang Provincial Experimental Animal Center Animal Welfare Ethics Committee (Ethics Number: ZJCLA-IACUC-20030066). We confirmed that all methods are reported in accordance with ARRIVE guidelines (https://arriveguidelines.org) for the reporting of animal experiments. Consent for publication Consent to publication-Not applicable. Competing interests The authors have no conflicts of interest to disclose. Fundings This study was supported by Natural Science Foundation of China (82205012) and Zhejiang Province Medical and Health Science and Technology Program (2022KY929,2021KY841). Authors contributions YW and WC designed the study. YW, WC and HZ carried out the in vitro experiments and wrote the original manuscript. YW, WC, HZ, JY and YL performed the in vivo experiments. All authors have read and approved the final version of the manuscript. Acknowledgements We thank the managers of the Animal Experiment Center of Zhejiang University of Chinese Medicine. Availability of data and material Summary statistics of COPD, IS and AMI were downloaded from the Medical Research Center-Integrative Epidemiology Unit (MRC-IEU) OpenGWAS database (https://gwas.mrcieu.ac.uk/) under accession no. finn-b-J10_COPD (COPD), ebi-a-GCST006907 (IS), and ukb-b-3469 (AMI). Data used to generate the main result are summarized in Supplementary Tables. The bidirectional MR analysis results, heterogeneity test (Q statistics) results, Egger intercept results, and MR-PRESSO global test results are summarized in eTables 2–5, respectively. Codes are available on reasonable request. All supporting data for in vivo and in vitro are included in this article and are available from the corresponding authors on reasonable request. References Agustí A, Celli BR, Criner GJ, Halpin D, Anzueto A, Barnes P, et al. Global Initiative for Chronic Obstructive Lung Disease 2023 Report: GOLD Executive Summary. Eur Respir J 2023; 61. Christenson SA, Smith BM, Bafadhel M, Putcha N. Chronic obstructive pulmonary disease. Lancet 2022; 399:2227–2242. Zhou M, Wang H, Zeng X, Yin P, Zhu J, Chen W, et al. Mortality, morbidity, and risk factors in China and its provinces, 1990–2017: a systematic analysis for the Global Burden of Disease Study 2017. Lancet 2019; 394:1145–1158. 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Exp Neurol 2023; 365:114426. Abo-El Fetoh ME, Abdel-Fattah MM, Mohamed WR, Ramadan LAA, Afify H. Cyclooxygenase-2 activates EGFR-ERK1/2 pathway via PGE2-mediated ADAM-17 signaling in testosterone-induced benign prostatic hyperplasia. Inflammopharmacology 2023; 31:499–516. Shi Q, Jiang Z, Yang J, Cheng Y, Pang Y, Zheng N, et al. A Flavonoid Glycoside Compound from Murraya paniculata (L.) Interrupts Metastatic Characteristics of A549 Cells by Regulating STAT3/NF-κB/COX-2 and EGFR Signaling Pathways. Aaps j 2017; 19:1779–1790. Fu L, Chen W, Guo W, Wang J, Tian Y, Shi D, et al. Berberine Targets AP-2/hTERT, NF-κB/COX-2, HIF-1α/VEGF and Cytochrome-c/Caspase Signaling to Suppress Human Cancer Cell Growth. PLoS One 2013; 8:e69240. Wang M, Dai T, Li S, Wang W. Eugenol suppresses the proliferation and invasion of TNF-α-induced fibroblast-like synoviocytes via regulating NF-κB and COX-2. Biochem Biophys Res Commun 2022; 612:63–69. Additional Declarations No competing interests reported. Supplementary Files supplementaryfigure1.tif Supplementary figure 1: Forest plot (A) and “leave-one-out” analysis plot (B) for MR analysis of the causal effect of COPD on IS. Forest plot (C) and “leave-one-out” analysis plot (D) for MR analysis of the causal effect of COPD on AMI. SupplementaryTables.xlsx Supplementary tables: Result data of MR results and reverse MR results WBofBaxintheaortatissue.tif WBofBaxinthecells.tif WBofBcl2intheaortatissue.tif WBofBcl2inthecells.tif WBofCOX2intheaortatissue.tif WBofCOX2inthecells.tif WBofGAPDHintheaortatissue1.tif WBofGAPDHintheaortatissue2.tif WBofGAPDHinthecells1.tif WBofGAPDHinthecells2.tif WBofcleavedcaspase3intheaortatissue.tif WBofcleavedcaspase3inthecells.tif WBofpp65inthecells.tif WBofpp65intheaortatissue.tif WBofp65intheaortatissue.tif WBofp65inthecells.tif 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. 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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-4384507","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":313882499,"identity":"96337bd2-5339-4e80-afda-51e6f51a9091","order_by":0,"name":"You Wu","email":"","orcid":"","institution":"The Second Affiliated Hospital of Zhejiang Chinese Medical University","correspondingAuthor":false,"prefix":"","firstName":"You","middleName":"","lastName":"Wu","suffix":""},{"id":313882500,"identity":"7cd0c114-1223-4f8f-ac64-c12120445db2","order_by":1,"name":"Houwen Zhang","email":"","orcid":"","institution":"The Second Affiliated Hospital of Zhejiang Chinese Medical University","correspondingAuthor":false,"prefix":"","firstName":"Houwen","middleName":"","lastName":"Zhang","suffix":""},{"id":313882501,"identity":"b3d6b22b-0105-49b5-8545-065423dadef4","order_by":2,"name":"Jialin Yu","email":"","orcid":"","institution":"The Second Affiliated Hospital of Zhejiang Chinese Medical University","correspondingAuthor":false,"prefix":"","firstName":"Jialin","middleName":"","lastName":"Yu","suffix":""},{"id":313882502,"identity":"24c3ba27-1beb-4351-b1a0-9e965cf78664","order_by":3,"name":"Yu Liang","email":"","orcid":"","institution":"The Second Affiliated Hospital of Zhejiang Chinese Medical University","correspondingAuthor":false,"prefix":"","firstName":"Yu","middleName":"","lastName":"Liang","suffix":""},{"id":313882503,"identity":"c0b9496b-f16d-437e-9776-9c62c90e21f9","order_by":4,"name":"Wanru Cai","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA3UlEQVRIie3QMQrCMBSA4YRAXAKucegdIoW6FL1KitAzdLMlUBfR1YKHKHiBlIBdCq6KSxY7eQA3tdZJJNXNIf/4eN/wHgA22x/WRzCWnNHZ6jWAcRcZzEWideTDLP6WsKoUQ12FMJffEnAIUhqkCrl7dd4S4Du5RLU2CbhuCfbkzjsRELq5xCNmIoi2hHjFAj+ICnJJMDUR/CLUFaQht25CSCEYr0LG8JPIbkJ7SaJ55HNaYe+4YVM3e9xlJBPV08WVUd5fqfpwicbOshS1kbzVvAr9sG+z2Wy2z90B0VVOPrH9ojMAAAAASUVORK5CYII=","orcid":"","institution":"The Second Affiliated Hospital of Zhejiang Chinese Medical University","correspondingAuthor":true,"prefix":"","firstName":"Wanru","middleName":"","lastName":"Cai","suffix":""}],"badges":[],"createdAt":"2024-05-07 17:09:38","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-4384507/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-4384507/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":58517200,"identity":"431af43d-b662-40c8-8302-567c62ccdb6e","added_by":"auto","created_at":"2024-06-17 16:54:45","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":134418,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eAssociation between COPD, IS and AMI estimated by the MR.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eChronic Obstructive Pulmonary Disease (COPD), ischemic stroke (IS), acute myocardial infarction (AMI), Mendelian randomization (MR). \u003cstrong\u003e(A)\u003c/strong\u003eThe effects of COPD on IS and AMI. \u003cstrong\u003e(B) \u003c/strong\u003eScatter plot showing the effect of genetic instruments on COPD risk against their effect on IS. \u003cstrong\u003e(C) \u003c/strong\u003eScatter plot showing the effect of genetic instruments on COPD risk against their effect on AMI.\u003c/p\u003e","description":"","filename":"Onlinefigure1.png","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/54f83ff30b227b19bd2b1b5d.png"},{"id":58517809,"identity":"16729a09-230c-4e7a-b79c-85fc1182c5e4","added_by":"auto","created_at":"2024-06-17 17:02:45","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":10766677,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe pulmonary function, blood lipids and pathological changes in lung tissue of mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(A) \u003c/strong\u003eFunctional residual capacity (FRC), resistance index (RI), dynamic lung compliance (Cdyn), and minute ventilation (MV); \u003cstrong\u003e(B) \u003c/strong\u003eSerum levels of total cholesterol (TC), triglycerides (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C);\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(C) \u003c/strong\u003ePathological changes observed in lung tissue. Data were presented as mean ± standard deviation. n=8, *P\u0026lt;0.05, ** P\u0026lt;0.01, ***P\u0026lt;0.001, ns = not significant. Magnification, 100 ×. Scale bar, 200 µm.\u003c/p\u003e","description":"","filename":"figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/a3b8e9a28cf08dddc26022d2.png"},{"id":58517807,"identity":"07d4fed3-4428-4520-ba26-2119ed2aa862","added_by":"auto","created_at":"2024-06-17 17:02:45","extension":"jpg","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":2220827,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe lipid accumulation, atherosclerotic plaques, expression of apoptosis-related proteins and key proteins of the F-κB/COX-2 signaling pathway in the aortic root of mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e(A) \u003c/strong\u003eThe lipid accumulation in the aorta; \u003cstrong\u003e(B)\u003c/strong\u003e The area of atherosclerotic plaque in the aortic root; \u003cstrong\u003e(C) \u003c/strong\u003eThe expression levels of cleaved-caspase 3, Bcl-2, and Bax in the aorta tissue; \u003cstrong\u003e(D)\u003c/strong\u003e The expression levels of p-p65, p65, and COX-2 in the aorta tissue; Data were presented as mean ± standard deviation. n=8, *P\u0026lt;0.05, ** P\u0026lt;0.01, ***P\u0026lt;0.001, ns = not significant.\u003c/p\u003e","description":"","filename":"figure3.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/ad5d24de5cd06df6b51ef017.jpg"},{"id":58518117,"identity":"df66fb61-3d8f-43bc-9d73-408aba1a8e98","added_by":"auto","created_at":"2024-06-17 17:10:45","extension":"jpg","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":1978976,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eThe levels of IL-6, IL-1β, TNF-α, PGE2, MCP-1, VCAM-1, and ICAM-1 were measured in the serum and aorta tissue of mice.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData were presented as mean ± standard deviation. n=8, *P\u0026lt;0.05, ** P\u0026lt;0.01, ***P\u0026lt;0.001, ns = not significant.\u003c/p\u003e","description":"","filename":"figure4.jpg","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/314aacded51e31c2bc51b32a.jpg"},{"id":58517210,"identity":"acd4dbb7-fc00-4c60-bd94-5bdd961e25e6","added_by":"auto","created_at":"2024-06-17 16:54:45","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":2571628,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eEvaluation of aortic endothelial cell migration ability, tube formation ability, expression levels of cellular inflammatory factors, changes in p65 translocation from the cytoplasm to the nucleus, and expression of proteins associated with the NF-κB/COX-2 signaling pathway.\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e(A) Represents the assessment of cell migration ability in each group; \u003cstrong\u003e(B)\u003c/strong\u003e Represents the assessment of cell tube formation ability in each group; \u003cstrong\u003e(C)\u003c/strong\u003e Represents the measurement of IL-6, IL-1β, TNF-α, PGE2, MCP-1, VCAM-1, and ICAM-1 levels in cells in each group; \u003cstrong\u003e(D)\u003c/strong\u003eRepresents the examination of p65 nuclear translocation in cells in each group; \u003cstrong\u003e(E)\u003c/strong\u003e Represents the analysis of p-p65, p65, and COX-2 expression levels in cells in each group. Data were presented as mean ± standard deviation. *P\u0026lt;0.05, ** P\u0026lt;0.01, ***P\u0026lt;0.001, ns = not significant.\u003c/p\u003e","description":"","filename":"Onlinefigure6.png","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/fdcc62be5ad985c8f41085a2.png"},{"id":63572240,"identity":"d7fa0bd9-137a-40de-a6a7-f06cb5146f7f","added_by":"auto","created_at":"2024-08-29 18:01:38","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":23399494,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/8cfcebb1-43cc-4557-b4ac-0c326e6e527e.pdf"},{"id":58517813,"identity":"85e1dbfe-07fb-4267-aa48-4de64febc3e5","added_by":"auto","created_at":"2024-06-17 17:02:46","extension":"tif","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":25139542,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary figure 1: \u003c/strong\u003eForest plot (A) and “leave-one-out” analysis plot (B) for MR analysis of the causal effect of COPD on IS. Forest plot (C) and “leave-one-out” analysis plot (D) for MR analysis of the causal effect of COPD on AMI.\u003c/p\u003e","description":"","filename":"supplementaryfigure1.tif","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/5b6786c8f42725a70737de00.tif"},{"id":58517202,"identity":"1a15facd-81a4-4580-884d-55ed9aed7c92","added_by":"auto","created_at":"2024-06-17 16:54:45","extension":"xlsx","order_by":2,"title":"","display":"","copyAsset":false,"role":"supplement","size":28150,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eSupplementary tables: \u003c/strong\u003eResult data of MR results and reverse MR results\u003c/p\u003e","description":"","filename":"SupplementaryTables.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/69ad8f210223e93b4ca38f4b.xlsx"},{"id":58517206,"identity":"e3d86a67-02fa-48b5-96b4-2b631a7edfd6","added_by":"auto","created_at":"2024-06-17 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16:54:46","extension":"tif","order_by":13,"title":"","display":"","copyAsset":false,"role":"supplement","size":369802,"visible":true,"origin":"","legend":"","description":"","filename":"WBofcleavedcaspase3intheaortatissue.tif","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/86f8205da35096df33d38975.tif"},{"id":58517221,"identity":"706deb83-9681-405a-a175-84da2c8b7fb6","added_by":"auto","created_at":"2024-06-17 16:54:47","extension":"tif","order_by":14,"title":"","display":"","copyAsset":false,"role":"supplement","size":369802,"visible":true,"origin":"","legend":"","description":"","filename":"WBofcleavedcaspase3inthecells.tif","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/9059e94548c5db3371cef6c3.tif"},{"id":58517814,"identity":"7948068e-0a28-4f81-957b-95dbd024675c","added_by":"auto","created_at":"2024-06-17 17:02:46","extension":"tif","order_by":15,"title":"","display":"","copyAsset":false,"role":"supplement","size":369802,"visible":true,"origin":"","legend":"","description":"","filename":"WBofpp65inthecells.tif","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/84f04da8e241397bd73a6d98.tif"},{"id":58517216,"identity":"d0c62b8f-0854-420b-ab3e-d79d54924285","added_by":"auto","created_at":"2024-06-17 16:54:46","extension":"tif","order_by":16,"title":"","display":"","copyAsset":false,"role":"supplement","size":369802,"visible":true,"origin":"","legend":"","description":"","filename":"WBofpp65intheaortatissue.tif","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/16fdbeccdab021b05df80851.tif"},{"id":58517213,"identity":"ce28e274-8a1b-4fea-a9bf-06dde597036d","added_by":"auto","created_at":"2024-06-17 16:54:46","extension":"tif","order_by":17,"title":"","display":"","copyAsset":false,"role":"supplement","size":369802,"visible":true,"origin":"","legend":"","description":"","filename":"WBofp65intheaortatissue.tif","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/835dfb50117ba5a6ac8f17f9.tif"},{"id":58517815,"identity":"2040a159-6a1b-415e-8424-d82cc95350f2","added_by":"auto","created_at":"2024-06-17 17:02:47","extension":"tif","order_by":18,"title":"","display":"","copyAsset":false,"role":"supplement","size":369802,"visible":true,"origin":"","legend":"","description":"","filename":"WBofp65inthecells.tif","url":"https://assets-eu.researchsquare.com/files/rs-4384507/v1/93896da625d2075d3ab991c4.tif"}],"financialInterests":"No competing interests reported.","formattedTitle":"The Causal Relationship between Chronic Obstructive Pulmonary Disease and Arterial Thrombotic Diseases: Role of Systemic Inflammation and NF- κB/COX-2 Pathway","fulltext":[{"header":"Introduction","content":"\u003cp\u003eChronic obstructive pulmonary disease (COPD) is a heterogeneous lung condition. \u003csup\u003e[\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]\u003c/sup\u003e It is a leading cause of death worldwide and poses a significant threat to public health. COPD often coexists with various diseases, particularly cardiovascular and cerebrovascular diseases. \u003csup\u003e[\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]\u003c/sup\u003e Numerous studies have suggested that COPD independently increases the risk of cardiovascular diseases. \u003csup\u003e[\u003cspan additionalcitationids=\"CR4\" citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]\u003c/sup\u003e Therefore, it is crucial to investigate the complex relationships and underlying mechanisms between COPD and cardiovascular disorders to enhance prevention, treatment, and overall management strategies.\u003c/p\u003e \u003cp\u003eCOPD involves the activation of multiple inflammatory cells and factors, which contribute to the inflammatory response. \u003csup\u003e[\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]\u003c/sup\u003e COPD patients experience persistent inflammation not only in the lungs but also throughout their entire system. \u003csup\u003e[\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e]\u003c/sup\u003e Some research has shown that this systemic inflammation increases the risk of various health issues, such as diabetes, stroke, cardiovascular diseases, lung cancer, and pneumonia, by 2\u0026ndash;4 times in COPD patients. \u003csup\u003e[\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]\u003c/sup\u003e Therefore, systemic inflammatory responses are considered significant contributors to the development and progression of these comorbidities in individuals with COPD. \u003csup\u003e[\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eIt is well-established that arteriosclerosis is a primary factor in the onset of myocardial infarction and stroke. A large body of compelling experimental and clinical data now indicates that inflammation participates fundamentally in atherogenesis (AS) and in the pathophysiology of ischaemic events. \u003csup\u003e[\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]\u003c/sup\u003e Research shows that NF-κB is one of the initiating mechanisms for vascular endothelial cell injury. \u003csup\u003e[\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]\u003c/sup\u003e Activated NF-κB can induce the continuous expression of cytokines, adhesion molecules, and enzymes related to the amplification of the inflammatory cascade. \u003csup\u003e[\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e]\u003c/sup\u003e Adhesion molecules such as monocyte chemoattractant protein-1 (MCP-1), intercellular cell adhesion molecule-1 (ICAM-1), vascular cellular adhesion molecule-1 (VCAM-1), and the selectin family are also expressed. \u003csup\u003e[\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]\u003c/sup\u003e NF-κB can increase the expression of inflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-8 (IL-8). Conversely, these inflammatory mediators can also promote the activation of NF-κB, thereby amplifying the inflammatory response. \u003csup\u003e[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e]\u003c/sup\u003e Upon activation, NF-κB also induces the enhanced expression of cyclooxygenase-2 (COX-2). Once activated, COX-2 can both reduce the bioavailability of nitric oxide (NO) and promote the accumulation of reactive oxygen species, intensifying oxidative stress. \u003csup\u003e[\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]\u003c/sup\u003e It can also catalyze the production of prostaglandin E2 (PGE2), activating inflammatory cells at the site of inflammation, inducing the release of chemokines, recruiting inflammatory cells, and inducing the production of multiple inflammatory factors, thereby maintaining and amplifying the inflammatory cascade. \u003csup\u003e[\u003cspan additionalcitationids=\"CR21\" citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e22\u003c/span\u003e]\u003c/sup\u003e These factors increase vascular permeability and uptake of oxidized low-density lipoprotein cholesterol (OxLDL) \u003csup\u003e[\u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e23\u003c/span\u003e]\u003c/sup\u003e promoting plaque formation and participating in the onset and development of AS. Studies have shown that by inhibiting the activity of NF-κB and suppressing the expression of COX-2 messenger RNA (mRNA), the progression of inflammation can be inhibited, providing a protective effect on endothelial cells. \u003csup\u003e[\u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e24\u003c/span\u003e, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e25\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eMendelian randomization (MR) analysis has emerged as a potent method for identifying causal relationships between risk factors and diseases, using genetic variants as instrumental variables (IVs). Building on this, the current study utilizes two-sample Mendelian randomization (TSMR) analysis to ascertain the potential causal link between COPD and the risk of developing cardiovascular and cerebrovascular diseases. Additionally, through both in vivo and in vitro experiments, we confirmed that systemic inflammation induced by COPD promotes atherogenesis lesions by activating the NF-κB/COX-2 pathway. \u003csup\u003e[\u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e26\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e"},{"header":"Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eMendelian Randomization Analysis\u003c/h2\u003e \u003cp\u003eThis study employed the TSMR approach, considering COPD as the exposure factor and both stroke and myocardial infarction as outcome variables to establish causal links. Data related to COPD, ischemic stroke (IS) \u003csup\u003e[\u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e27\u003c/span\u003e]\u003c/sup\u003e, and acute myocardial infarction (AMI) were respectively sourced from FinnGen Biobank, MEGASTROKE, and the MRC-IEU organizations (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCharacteristics of data sources and strength of IVs used in the Mendelian randomization study\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"7\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eExposures/Outcomes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGWAS ID\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eConsortium\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEthnicity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eSample Sizes\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNumber of SNPs\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eYear\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCOPD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003efinn-b-J10_COPD\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eFinnGen Biobank\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEuropean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e193,638\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16,380,382\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e2021\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eIschemic stroke\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eebi-a-GCST006907\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMEGASTROKE\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEuropean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e150,765\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e8,418,349\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e2018\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAcute myocardial\u003c/p\u003e \u003cp\u003einfarction\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eukb-b\u0026minus;3469\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMRC-IEU\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eEuropean\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e463,010\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e9,851,867\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e2018\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"7\"\u003eGWAS:Genome-Wide Association Studies,COPD:Chronic obstructive pulmonary disease,SNPs: single nucleotide polymorphisms, NA: Not Available.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eTo conduct the analysis, we compiled single nucleotide polymorphisms (SNPs) associated with COPD at a significant level of P\u0026thinsp;\u0026lt;\u0026thinsp;5 \u0026times; 10\u003csup\u003e\u0026minus;\u0026thinsp;6\u003c/sup\u003e. In order to ensure the independence of these SNPs, we established a linkage disequilibrium threshold (r\u003csup\u003e2\u003c/sup\u003e) of 0.01 and a genetic distance of 5000 kb for the selection process. We identified the phenotypes associated with the remaining SNPs using the Human Genotype-Phenotype Association Database (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ewww.phenoscanner.medschl.cam.ac.uk\u003c/a\u003e\u003c/span\u003e\u003cspan address=\"http://www.phenoscanner.medschl.cam.ac.uk\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e), \u003csup\u003e[\u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e28\u003c/span\u003e]\u003c/sup\u003e with a special emphasis on excluding SNPs whose corresponding phenotypes were significantly associated with the outcomes. The datasets for both the exposure and outcome variables were combined, while palindromic SNPs were removed to ensure the integrity of the analysis. In the meanwhile, to mitigate interference from reverse causality, reverse MR analysis was employed using SNPs related to IS and AMI.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec4\" class=\"Section2\"\u003e \u003ch2\u003eMice\u003c/h2\u003e \u003cp\u003e8-week-old male SPF grade ApoE knockout mice (ApoE\u0026minus;/\u0026minus;) were purchased from the Experimental Animal Center of Hangzhou Medical College in Zhejiang, China (Licence Number: SYXK(Zhe)2019-0011) and housed in a specific pathogen-free (SPF) facility in a controlled environment. All experimental protocols were approved by the Zhejiang Provincial Experimental Animal Center Animal Welfare Ethics Committee (Ethics Number: ZJCLA-IACUC-20030066). All methods were carried out in accordance with relevant guidelines and regulations. All animal experiments were taken place in SPF Animal Laboratory at Zhejiang Chinese Medical University. The mice were randomly divided into the following groups:\u003c/p\u003e \u003cp\u003eControl group: Mice were given a normal diet, and equivalent doses of saline were instilled during the PPE and LPS induction for the COPD model.\u003c/p\u003e \u003cp\u003eCOPD group: Mice were instilled with 1.2 IU of PPE via the airway once a week for a total of 4 times. Two weeks after the final PPE instillation, 200 \u0026micro;g of LPS was instilled once a week, for a total of 2 times.\u003c/p\u003e \u003cp\u003eHFD group: To establish the AS model, mice were fed a HFD (comprising 3% cholesterol, 0.5% sodium cholate, 0.2% propylthiouracil, 5% sugar, 10% lard, and 81.3% base diet) for 7 weeks.\u003c/p\u003e \u003cp\u003eHFD\u0026thinsp;+\u0026thinsp;COPD group: Alongside initiating HFD, COPD interventions were given as well. Mice received 1.2 IU of PPE instilled via the airway once a week for a total of 4 times. One week after the last PPE instillation, 200 \u0026micro;g of LPS was instilled once a week for a total of 2 times.\u003c/p\u003e \u003cp\u003eHFD\u0026thinsp;+\u0026thinsp;COPD\u0026thinsp;+\u0026thinsp;BAY11-7082 group: While establishing the combined COPD and AS model, mice were given intraperitoneal injections of the NF-κB inhibitor BAY11-7082 (HY-13453, MedChemExpress) (5 mg/kg) on the same day as the PPE instillation. Subsequently, injections were given every other day until the end of the experiment.\u003c/p\u003e \u003cp\u003eHFD\u0026thinsp;+\u0026thinsp;COPD\u0026thinsp;+\u0026thinsp;NS-398 group: While establishing the combined COPD and AS model, mice were given intraperitoneal injections of the COX-2 inhibitor NS-398 (HY-13913, MedChemExpress) (5 mg/kg) on the same day as the PPE instillation. Subsequently, injections were given every other day until the end of the experiment.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section2\"\u003e \u003ch2\u003eCells\u003c/h2\u003e \u003cp\u003ePrimary mouse aorta endothelial cells were used for this study. The aortic endothelial cells were then purified using anti-CD31 coupled magnetic beads. \u003csup\u003e[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e]\u003c/sup\u003e \u003cb\u003e(\u003c/b\u003eFig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e5\u003c/span\u003eA-B\u003cb\u003e)\u003c/b\u003e Simultaneously, serum was extracted from both the COPD group of mice and the control group of mice. The cells were randomly divided into the following groups:\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eControl group: cells received no special treatment.\u003c/p\u003e \u003cp\u003eNS group: cells received serum from the control group mice.\u003c/p\u003e \u003cp\u003eCS group: cells received serum from COPD mice.\u003c/p\u003e \u003cp\u003eCS\u0026thinsp;+\u0026thinsp;BAY11-7082 group: cells received BAY11-7082 while receiving serum from COPD mice.\u003c/p\u003e \u003cp\u003eCS\u0026thinsp;+\u0026thinsp;BAY11-7082\u0026thinsp;+\u0026thinsp;oe-NC group: cells received serum from COPD mice, BAY11-7082, and transfected empty plasmid simultaneously.\u003c/p\u003e \u003cp\u003eCS\u0026thinsp;+\u0026thinsp;BAY11-7082\u0026thinsp;+\u0026thinsp;oe-COX-2 group: cells received serum from COPD mice, BAY11-7082, and transfected COX-2 expressing plasmid (purchased from Santa Cruz Biotechnology) simultaneously.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec6\" class=\"Section2\"\u003e \u003ch2\u003ePulmonary Function Test\u003c/h2\u003e \u003cp\u003eMice were anaesthetized by intraperitoneal injection using a 3% pentobarbital sodium solution. After the tracheal cannula was connected to the airway of the plethysmograph (PFT, DSI), the measurements of functional residual capacity (FRC), resistance index (RI), dynamic lung compliance (Cdyn), and minute ventilation (MV) were taken. At the end of these experiments, all mice were euthanized by CO\u003csub\u003e2\u003c/sub\u003e asphyxiation.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003eMice Lipid Profile\u003c/h2\u003e \u003cp\u003eThe following mouse kits were used: Total Cholesterol (TC), Triglyceride (TG), High-Density Lipoprotein Cholesterol (HDL-C), and Low-Density Lipoprotein Cholesterol (LDL-C) (A111-1-1, A110-1-1, A112-1-1 and A113-1-1, Nanjing Jiancheng Bioengineering Institute). These serum samples then underwent testing on a fully-automated biochemistry analyser (COBAS INTEGRA 800, Roche, Switzerland).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eMice Tissue\u003c/h2\u003e \u003cp\u003eThe lung tissue and aortic roots of the different groups of mice were evaluated using Hematoxylin and Eosin (H\u0026amp;E) (C0105S, Beyotime) staining to assess the pathological changes in lung tissues and the size of arteriosclerotic plaques. Additionally, Oil Red O (G1015-100ML, Servicebio) staining was employed to evaluate lipid accumulation in the aorta.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section2\"\u003e \u003ch2\u003eCells Functions\u003c/h2\u003e \u003cp\u003eCells were seeded in a 96-well plate, treated with CCK-8 (CA1210, Solarbio) solution, and absorbance at 450 nm was measured to determine viability. Apoptosis was evaluated using flow cytometry (Attune\u0026trade; NxT, Thermo Fisher Scientific), where cells were stained with Annexin V-FITC (C1062S, Beyotime) and Propidium Iodide (PI), and apoptotic cells were detected based on fluorescence microscope (M205 FCA, Leica). Cells were seeded in Matrix-Gel\u0026trade; chambers, cultured, and stained with crystal violet for migration assessment. Tubule formation potential was investigated by starving cells, seeding them in Matrigel-coated wells, and observing the formation of tube networks.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eRT-qPCR\u003c/h2\u003e \u003cp\u003eTotal RNA was extracted from the cells using TRIzol reagent. cDNA was then synthesized using PrimeScript reverse transcriptase following the manufacturer's protocol. The amplified products were COX-2 forward primer: 5'-AGGACTCTGCTCACGAAGGA-3', COX-2 reverse primer: 5'- TGACATGATTGGAACAGCA-3', GAPDH forward primer: 5'-ACCCTTAAGAGGATGCTGC-3', and GAPDH reverse primer: 5'- CCCAATACGGCCAAATCCGT-3'. The gene expression levels were quantified using the 2^-ΔΔCQ method and normalized to the internal reference gene GAPDH.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eELISA\u003c/h2\u003e \u003cp\u003eThe relevant inflammatory factors in the mice aortic tissues, serum, and cells from each group were detected using enzyme-linked immunosorbent assay (ELISA) kits for IL-6, IL-1β, TNF-α, PGE2, MCP-1, VCAM-1, and ICAM-1(SEKM-0007, SEKM-0002, SEKM-0034, SEKM-0173, SEKM-0108, SEKM-0037, SEKM-0132). Analyses were performed according to the kit instructions.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eWestern Blot\u003c/h2\u003e \u003cp\u003eProteins were extracted from the aortic tissues of various mouse groups and from the cells of each group using RIPA lysis buffer (D3910201, Sigma) supplemented with 1% Phenylmethanesulfonyl fluoride (PMSF). Proteins were then separated by Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE), with an equal amount of protein (30\u0026micro;g) loaded per lane, and subsequently transferred onto PVDF membranes (IPVH00010, Millipore). These membranes were incubated overnight at 4\u0026deg;C with antibodies diluted at 1:3000 for p-p65, p65, COX-2, caspase 3, Bcl-2, Bax, and GAPDH (3033, 8242, 4842, 9662, 3498, 2772 and 5174, CST). This was followed by a 1-hour incubation with Horseradish Peroxidase (HRP) conjugated secondary antibodies. Densitometric analysis was conducted using ImageJ software.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003e4. Statistical Analysis\u003c/h2\u003e \u003cp\u003eWe utilized the software packages \"TwoSampleMR\" and \"MRPRESSO\" with R version 4.3.1 to conduct MR analysis. We utilized several methods including inverse variance weighting (IVW), \u003csup\u003e[\u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e30\u003c/span\u003e, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e31\u003c/span\u003e]\u003c/sup\u003e MR-Egger, \u003csup\u003e[\u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e32\u003c/span\u003e, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]\u003c/sup\u003e Simple mode, weighted mode and weighted median method. \u003csup\u003e[\u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e34\u003c/span\u003e]\u003c/sup\u003e The IVW method was used as the primary analysis. We assessed heterogeneity by employing Cochran\u0026rsquo;s Q method, considering heterogeneity to be significant at p\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003csup\u003e[\u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e35\u003c/span\u003e]\u003c/sup\u003e We utilized the MR-Egger intercept test \u003csup\u003e[\u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e33\u003c/span\u003e]\u003c/sup\u003e and the MR-PRESSO global test to identify and remove any potential outliers with horizontal pleiotropic effects that might have significantly influenced the estimation results. \u003csup\u003e[\u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e36\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThis study utilized SPSS 22.0 software for the in vivo and in vitro experiments statistical analysis. Quantitative data were assessed for normality and presented as the mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (x̅ \u0026plusmn; SD). The T test was employed to analyse differences between groups, and statistical significance was determined when P\u0026thinsp;\u0026lt;\u0026thinsp;0.05. Graphs were generated using GraphPad Prism 8.0 (GraphPad, USA).\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eMR Analysis\u003c/h2\u003e \u003cp\u003eIn our MR analysis investigating the impact of COPD on cardiovascular and cerebrovascular diseases, a meticulous selection of 19 SNPs associated with the influence of COPD on IS, and 7 SNPs associated with AMI was conducted (\u003cb\u003eSupplementary Tables\u003c/b\u003e). Using the random-effects IVW method, a significant positive association was observed between COPD and the risk of IS (OR: 1.152, 95% CI: 1.022-1.300, P\u0026thinsp;=\u0026thinsp;0.021) and AMI (OR: 1.001, 95% CI: 1.001\u0026ndash;1.002, P\u0026thinsp;=\u0026thinsp;0.029) (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e1\u003c/span\u003e). These findings were consistently supported across various methodologies. Sensitivity analyses, encompassing MR-Egger intercepts and Cochran Q tests, did not reveal significant directional pleiotropy, heterogeneity, or notable outliers, further substantiating the robustness of our results (\u003cb\u003eSupplementary Tables\u003c/b\u003e). Moreover, no single SNP was found to disproportionately influence the overall causal estimates, as confirmed by the leave-one-out analysis (\u003cb\u003eSupplementary Fig.\u0026nbsp;1\u003c/b\u003e), thereby underscoring the stability and reliability of our study outcomes. For the reverse MR, no significant effect of IS and AMI on COPD was observed (\u003cb\u003eSupplementary Tables\u003c/b\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eEnhanced Pulmonary Dysfunction and Atherogenesis in COPD and HFD Mice\u003c/h2\u003e \u003cp\u003eOur investigation revealed significant functional alterations in the pulmonary system in different mouse models. In both the COPD and HFD groups, there was a noticeable increase in FRC and RI, along with a significant decrease in Dynamic Lung Cdyn and MV, compared to the control group (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003eA-D). The decline in lung function was even more prominent in the HFD\u0026thinsp;+\u0026thinsp;COPD group.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003eHistological Analysis of Lung and Aortic Tissues\u003c/h2\u003e \u003cp\u003eIn the lung tissues, while the control group displayed intact alveolar structures, the COPD and HFD groups showed notable reductions in alveolar number accompanied by substantial macrophage infiltration (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003eI). This degeneration was even more pronounced in the HFD\u0026thinsp;+\u0026thinsp;COPD group. Interestingly, treatment with NF-κB and COX-2 inhibitors, particularly in the HFD\u0026thinsp;+\u0026thinsp;COPD\u0026thinsp;+\u0026thinsp;BAY11-7082 and HFD\u0026thinsp;+\u0026thinsp;COPD\u0026thinsp;+\u0026thinsp;NS-398 groups, led to a significant recovery in alveolar count and structural integrity.\u003c/p\u003e \u003cp\u003eIn the aortic tissues, changes were equally remarkable. The control group showed minimal atherosclerotic changes, whereas the HFD group exhibited a noticeable increase in atheromatous plaque formation. This atherosclerotic progression was even more extensive in the HFD\u0026thinsp;+\u0026thinsp;COPD group, with larger plaque formations (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eA \u003cb\u003eand B\u003c/b\u003e). However, in the groups treated with NF-κB and COX-2 inhibitors, there was a notable reduction in the size and complexity of atherosclerotic plaques.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eLipid Profile Alterations and Inflammatory Marker Dynamics\u003c/h2\u003e \u003cp\u003eLipid profile analysis revealed a significant dysregulation in the HFD group, with elevated levels of TC, TG and LDL-C, coupled with a decrease in HDL-C (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e2\u003c/span\u003eE-H). This dysregulation was exacerbated in the HFD\u0026thinsp;+\u0026thinsp;COPD group, suggesting that COPD may intensify lipid abnormalities in atherosclerosis. Inflammatory markers such as IL-6, IL-1β, TNF-α, PGE2, MCP-1, VCAM-1, and ICAM-1 displayed a significant increase in the COPD and HFD groups (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e4\u003c/span\u003e). This increase was even more pronounced in the HFD\u0026thinsp;+\u0026thinsp;COPD group, indicating an enhanced inflammatory response due to the combination of COPD and atherosclerosis.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec19\" class=\"Section2\"\u003e \u003ch2\u003eApoptosis and NF-κB/COX-2 Pathway Activation in Aortic Tissues\u003c/h2\u003e \u003cp\u003eWestern Blot analysis of aortic tissues from the COPD and HFD groups revealed an upregulation of apoptosis-related proteins Cleaved-caspase 3 and Bax, alongside a downregulation of Bcl-2, with the most significant changes observed in the HFD\u0026thinsp;+\u0026thinsp;COPD group (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eC \u003cb\u003eand D\u003c/b\u003e). The activation of the NF-κB/COX-2 pathway was also apparent in these groups, as demonstrated by elevated expressions of p-65, p-p65 and COX-2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e3\u003c/span\u003eE \u003cb\u003eand F\u003c/b\u003e). These molecular changes were significantly altered upon treatment with NF-κB and COX-2 inhibitors.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec20\" class=\"Section2\"\u003e \u003ch2\u003eEndothelial Cell Function Under COPD-Induced Systemic Inflammation\u003c/h2\u003e \u003cp\u003eIn vitro experiments utilizing primary aortic endothelial cells exposed to serum from COPD mice (CS group) revealed a reduction in cell viability and an increase in apoptosis rates compared to cells exposed to control serum (NS group). The application of BAY11-7082, an NF-κB inhibitor, markedly improved cell viability and reduced apoptosis rates (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e5\u003c/span\u003eC-F). However, overexpression of COX-2 in these cells negated the protective effects of BAY11-7082.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec21\" class=\"Section2\"\u003e \u003ch2\u003eMigration, Tubule Formation, and Inflammatory Responses in Endothelial Cells\u003c/h2\u003e \u003cp\u003eCell migration and tubule formation capabilities were assessed to understand the effects of COPD-induced systemic inflammation on endothelial cell function. A significant reduction in cell migration (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eA) and tubule formation (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eB) was observed in the CS group, which was reversed upon treatment with BAY11-7082. However, this reversal was diminished with COX-2 overexpression. Additionally, inflammatory markers in these cells showed an increase in the CS group, which was reduced following treatment with BAY11-7082 (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eC). Yet, this reduction was counteracted by COX-2 overexpression.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003eNF-κB/COX-2 Pathway Influence in Endothelial Cells\u003c/h2\u003e \u003cp\u003eDetailed analysis through immunofluorescence (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eD) and Western Blot (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003eE \u003cb\u003eand F\u003c/b\u003e) techniques revealed that exposure to COPD-induced serum led to enhanced nuclear translocation of p65 and increased expressions of p-p65 and COX-2 in endothelial cells. These molecular changes were attenuated by the NF-κB inhibitor BAY11-7082 but were reinstated upon COX-2 overexpression.\u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eIn this study, we utilized a TSMR approach to investigate the causal relationship between COPD and the increased risks of IS and AMI. Our analysis, which involved carefully selecting SNPs from large-scale biobanks, revealed a significant genetic correlation between COPD and these cardiovascular conditions. Additionally, we conducted complementary in vivo and in vitro experiments to further elucidate the role of systemic inflammation, specifically through the NF-κB/COX-2 pathway, in mediating this relationship.\u003c/p\u003e \u003cp\u003eMultiple studies have consistently demonstrated elevated levels of proinflammatory cytokines, including serum C-reactive protein, plasma fibrinogen, serum amyloid A, TNF-α, IL-6, IL-8, COX-2, etc. in patients with COPD. \u003csup\u003e[\u003cspan additionalcitationids=\"CR38 CR39 CR40\" citationid=\"CR37\" class=\"CitationRef\"\u003e37\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e]\u003c/sup\u003e This systemic inflammation is further exacerbated during acute exacerbations of COPD. \u003csup\u003e[\u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e29\u003c/span\u003e, \u003cspan additionalcitationids=\"CR42\" citationid=\"CR41\" class=\"CitationRef\"\u003e41\u003c/span\u003e\u0026ndash;\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e43\u003c/span\u003e]\u003c/sup\u003e In our COPD model, we also observed a corresponding increase in these inflammatory markers. However, the exact mechanism underlying this phenomenon remains unclear, possibly attributed to the leakage of various proinflammatory markers from the lungs into the bloodstream. These findings provide evidence for the presence of a systemic inflammatory response in individuals with COPD. \u003csup\u003e[\u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e44\u003c/span\u003e, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e45\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eThe NF-κB/COX-2 inflammatory pathway plays a crucial role in regulating AS. Gareus et al. \u003csup\u003e[\u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e46\u003c/span\u003e]\u003c/sup\u003e conducted research demonstrating that the depletion of key signaling molecules in the NF-κB pathway, namely nuclear factor κB kinase γ (IKKγ) or nuclear factor κB inhibitor α (IkappaBalpha, IκBα), significantly reduced the formation of atherosclerotic plaques in ApoE\u0026minus;/\u0026minus; mice. Additionally, circulating microRNAs (miRNAs) have been found to reduce inflammation and lesions in ApoE\u0026minus;/\u0026minus; atherosclerotic mice by inhibiting the activation of NF-κB in vascular endothelial cells. \u003csup\u003e[\u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e47\u003c/span\u003e]\u003c/sup\u003e This research suggests that the key vault protein can suppress the NF-κB signal-mediated inflammatory pathway, thereby inhibiting obesity and the development of AS. \u003csup\u003e[\u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e48\u003c/span\u003e]\u003c/sup\u003e Moreover, methyltransferase-like 14 (Mettl14) has been implicated in the inflammatory response of macrophages in AS through the NF-κB/IL-6 signaling pathway. \u003csup\u003e[\u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e49\u003c/span\u003e]\u003c/sup\u003e Studies have demonstrated that inhibiting NF-κB activity to suppress the expression of COX-2 mRNA can reduce oxidative stress, inhibit inflammation, and protect endothelial cells. \u003csup\u003e[\u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e50\u003c/span\u003e, \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e51\u003c/span\u003e]\u003c/sup\u003e Similar results were obtained when separately inhibiting NF-κB and COX-2.\u003c/p\u003e \u003cp\u003eAlthough the NF-κB/COX-2 signaling pathway has not been validated in the study of COPD combined with AS, it has been shown to have a significant impact on other diseases, particularly those related to inflammation. Research has demonstrated that inflammatory factors stimulate the translocation of NF-κB in human bronchial smooth muscle cells, and the activation of NF-κB in the airway epithelium leads to a significant increase in inflammatory factors. \u003csup\u003e[\u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e52\u003c/span\u003e]\u003c/sup\u003e The NF-κB signaling pathway plays a crucial role in regulating inflammation through COX-2 transcription. For example, Seyyed and colleagues' research indicates that NF-κB-induced COX-2 is an important factor in causing allergic asthma lung inflammation. \u003csup\u003e[\u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e53\u003c/span\u003e]\u003c/sup\u003e In a mouse model of LPS-induced liver injury, lycopene has been shown to alleviate liver function damage by inhibiting NF-κB/COX-2 signal transmission and reducing the expression of inflammatory factors in mouse serum.\u003csup\u003e[\u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e54\u003c/span\u003e]\u003c/sup\u003e In a rat model of pharyngitis, the NF-κB/COX-2 signaling pathway was significantly activated, and inhibiting the activity of this pathway alleviated pharyngitis-related throat symptoms. \u003csup\u003e[\u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e55\u003c/span\u003e]\u003c/sup\u003e\u003c/p\u003e \u003cp\u003eHE staining revealed increased inflammation indicators in arterial tissue and elevated inflammatory factors in serum of the HFD mouse group, accompanied by a decrease in the number of alveoli and minor macrophage infiltration within the alveoli. This may be related to high-fat diet and further experiments are needed to verify. \u003csup\u003e[\u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e56\u003c/span\u003e, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e57\u003c/span\u003e]\u003c/sup\u003e In two intervention experiments and treatments overexpressing COX-2, we noted a clear upstream-downstream relationship between NF-κB and COX-2 but have yet to investigate how to further enhance the expression of inflammatory factors subsequent to COX-2 expression. One potential mechanism could be that COX-2 synthesized PGE2, after binding with its receptors EP2 and EP4, activates adenylyl cyclase in cells causing increased intracellular cAMP levels. \u003csup\u003e[\u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e58\u003c/span\u003e]\u003c/sup\u003e Heightened cAMP levels further stimulate protein kinase A (PKA), which can phosphorylate and activate the transcription factor CREB (cAMP response element binding protein), promoting gene transcription of inflammation factors such as IL-6 and TNF-α. Additionally, PGE2 can promote the gene transcription of inflammatory factors such as IL-6 and TNF-α by activating the MAPK signaling pathway (including ERK, JNK and p38 kinases). \u003csup\u003e[\u003cspan citationid=\"CR59\" class=\"CitationRef\"\u003e59\u003c/span\u003e, \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e60\u003c/span\u003e]\u003c/sup\u003e The NF-κB/COX-2 inflammatory pathway presents an opportunity for targeting the prevention of COPD in combination with AS and the occurrence of malignant ischemic events. However, the safety and effectiveness of potential drugs such as Murraya paniculata, \u003csup\u003e[\u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e61\u003c/span\u003e]\u003c/sup\u003e Berberine Targets, \u003csup\u003e[\u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e62\u003c/span\u003e]\u003c/sup\u003e Eugenol, \u003csup\u003e[\u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e63\u003c/span\u003e]\u003c/sup\u003e etc. still require further verification through clinical practice.\u003c/p\u003e"},{"header":"Conclusions","content":"\u003cp\u003eThe convergence of genetic, clinical, and molecular data in our study provides strong evidence supporting the integration of arterial thrombotic disease risk management in the treatment and overall care strategies for patients with COPD. Furthermore, our findings suggest that targeting the NF-κB/COX-2 pathway could be a potential therapeutic approach to mitigate the increased arteriosclerosis and thrombosis formation risks associated with COPD, opening up new avenues for treatment strategies. In conclusion, this study improves our understanding of COPD as a systemic disease with significant cardiovascular implications. It highlights the importance of a comprehensive approach to managing COPD, addressing both pulmonary and cardiovascular aspects, in order to achieve better patient outcomes.\u003c/p\u003e "},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthics approval\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eData for the Mendelian andomization analysis was sourced from publicly available databases, negating the need for ethical approval or patient consent for this analysis. All animal experiments were taken place in SPF Animal Laboratory at Zhejiang Chinese Medical University and were in compliance with the ethical requirements of the Zhejiang Provincial Experimental Animal Center Animal Welfare Ethics Committee (Ethics Number: ZJCLA-IACUC-20030066).\u003c/p\u003e\n\u003cp\u003eWe confirmed that all methods are reported in accordance with ARRIVE guidelines (https://arriveguidelines.org) for the reporting of animal experiments.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eConsent to publication-Not applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCompeting interests\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors have no conflicts of interest to disclose.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFundings\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis study was supported by Natural Science Foundation of China (82205012) and\u0026nbsp;Zhejiang Province Medical and Health Science and Technology Program (2022KY929,2021KY841).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors contributions\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eYW and WC designed the study. YW, WC and HZ carried out the in vitro experiments and wrote the original manuscript. YW, WC, HZ, JY and YL performed the in vivo experiments. All authors have read and approved the final version of the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe thank the managers of the Animal Experiment Center of Zhejiang University of Chinese Medicine.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAvailability of data and material\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eSummary statistics of COPD, IS and AMI were downloaded from the Medical Research Center-Integrative Epidemiology Unit (MRC-IEU) OpenGWAS database (https://gwas.mrcieu.ac.uk/) under accession no. finn-b-J10_COPD (COPD), ebi-a-GCST006907 (IS), and ukb-b-3469 (AMI). Data used to generate the main result are summarized in Supplementary Tables. The bidirectional MR analysis results, heterogeneity test (Q statistics) results, Egger intercept results, and MR-PRESSO global test results are summarized in eTables 2\u0026ndash;5, respectively. Codes are available on reasonable request.\u003c/p\u003e\n\u003cp\u003eAll supporting data for in vivo and in vitro are included in this article and are available from the corresponding authors on reasonable request.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAgust\u0026iacute; A, Celli BR, Criner GJ, Halpin D, Anzueto A, Barnes P, et al. Global Initiative for Chronic Obstructive Lung Disease 2023 Report: GOLD Executive Summary. Eur Respir J 2023; 61.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eChristenson SA, Smith BM, Bafadhel M, Putcha N. Chronic obstructive pulmonary disease. 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Biochem Biophys Res Commun 2022; 612:63\u0026ndash;69.\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":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Chronic obstructive pulmonary disease, inflammation, nuclear factor-κB, cyclooxygenase-2, Mendelian Randomization","lastPublishedDoi":"10.21203/rs.3.rs-4384507/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-4384507/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003eBackground:\u003c/strong\u003e Chronic Obstructive Pulmonary Disease (COPD) is a significant global health issue that often coexists with arterial thrombotic diseases. This study aims to investigate the causal relationship between COPD and these diseases, focusing on the role of systemic inflammation and the NF-κB/COX-2 pathway.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods:\u003c/strong\u003e The Two-Sample Mendelian Randomization (TSMR) approach was used to analyze the genetic correlation between COPD and the risks of ischemic stroke (IS) and acute myocardial infarction (AMI) using data from several large biobanks. Additionally, in vivo experiments with ApoE knockout mice and in vitro assays with primary mouse aorta endothelial cells were conducted to explore the role of the NF-κB/COX-2 pathway in COPD-related systemic inflammation.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e The MR analysis revealed a significant association between COPD and increased risks of IS (OR: 1.152) and AMI (OR: 1.001). In vivo findings showed exacerbated pulmonary dysfunction and atherogenesis in mice with both COPD and high-fat diet (HFD), with notable histological changes in lung and aortic tissues. Inflammatory markers and lipid profiles were significantly altered in these models. In vitro studies demonstrated that COPD-induced systemic inflammation impaired endothelial cell function. These changes were mitigated by inhibiting the NF-κB/COX-2 pathway.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusions:\u003c/strong\u003e This study provides strong evidence of a causal link between COPD and an elevated risk of arterial thrombotic diseases, mediated by systemic inflammation and the NF-κB/COX-2 pathway. These findings highlight the importance of addressing arteriosclerosis and thrombosis formation risks in COPD management and suggest that the NF-κB/COX-2 pathway could be a potential therapeutic target for reducing comorbidity in COPD patients.\u003c/p\u003e","manuscriptTitle":"The Causal Relationship between Chronic Obstructive Pulmonary Disease and Arterial Thrombotic Diseases: Role of Systemic Inflammation and NF- κB/COX-2 Pathway","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2024-06-17 16:54:40","doi":"10.21203/rs.3.rs-4384507/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"7a94b7a7-c401-44b0-8ed1-8b9f96ef5b45","owner":[],"postedDate":"June 17th, 2024","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":33191356,"name":"Biological sciences/Genetics"},{"id":33191357,"name":"Biological sciences/Immunology"},{"id":33191358,"name":"Biological sciences/Neuroscience"},{"id":33191359,"name":"Health sciences/Diseases/Cardiovascular diseases"},{"id":33191360,"name":"Health sciences/Diseases/Neurological disorders"},{"id":33191361,"name":"Health sciences/Diseases/Respiratory tract diseases"}],"tags":[],"updatedAt":"2024-08-29T17:53:17+00:00","versionOfRecord":[],"versionCreatedAt":"2024-06-17 16:54:40","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-4384507","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-4384507","identity":"rs-4384507","version":["v1"]},"buildId":"qtupq5eGEP_6zYnWcrvyt","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}