BACKGROUND Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by inflammation and joint stiffness, finally leading to tissue destruction. Connective tissue growth factor (CTGF) is a critical factor in RA progression, which promotes fibroblast-like synoviocyte (FLS) proliferation, pannus formation, and the damage of cartilage as well as bone. Resolvin D1 (RvD1) can promote inflammation resolution in acute inflammatory diseases, and recently, effects of RvD1 on chronic inflammatory diseases also attracted attention. This study aimed to examine the effect of RvD1 on pannus formation in RA and the underlying mechanism. METHODS Serum levels of RvD1 and CTGF were determined in RA patients and healthy persons by UPLC-MS/MS and ELISA respectively. The levels of CTGF and inflammatory factors were assessed by qRT-PCR and ELISA. MicroRNA expression profile was determined by miRNA microarray. The effects of CTGF, RvD1, and miR-146a-5p on angiogenesis were evaluated with tube formation and chick chorioallantoic membrane (CAM) assays. Collagen-induced arthritis (CIA) mice were constructed to detect the effects of RvD1 and miR146a-5p on RA. STAT3 activation was determined by Western blotting. RESULTS RvD1 levels decreased while CTGF levels increased in RA patients' serum, and an inverse correlation of the concentrations of RvD1 and CTGF in the serum of RA patients was synchronously observed. In CIA mice, RvD1 suppressed angiopoiesis and decreased the expression of CTGF. Simultaneously, RvD1 significantly decreased CTGF and pro-inflammation cytokines levels in RA FLS. Furthermore, CTGF suppressed angiopoiesis and RvD1 inhibited the proliferation and migration of RA FLS and angiopoiesis. MiRNA microarray and qRT-PCR results showed that RvD1 upregulated miRNA-146a-5p. The transfection experiments demonstrated that miRNA-146a-5p could decrease inflammatory factors and CTGF levels. Moreover, miRNA-146a-5p decreased the proliferation of FLS and angiogenesis in vivo. MiRNA-146a-5p also suppressed angiogenesis and downregulated the expression of CTGF in CIA mice. Finally, Western blot results revealed that miRNA-146a-5p inhibited the activation of STAT3. CONCLUSION RvD1 is prone to alleviate RA progression through the upregulation of miRNA-146a-5p to suppress the expression of CTGF and inflammatory mediators, thereby decreasing pannus formation and cartilage damage.

中文翻译：

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Resolvin D1通过减少类风湿关节炎中miRNA-146a-5p的上调引起的结缔组织生长因子来抑制pan的形成。

背景技术类风湿关节炎（RA）是一种慢性自身免疫性疾病，其特征在于炎症和关节僵硬，最终导致组织破坏。结缔组织生长因子（CTGF）是RA进展的关键因素，可促进成纤维样滑膜细胞（FLS）增殖，血管pan形成以及软骨和骨骼的损伤。Resolvin D1（RvD1）可以促进急性炎症疾病的炎症消退，最近，RvD1对慢性炎症疾病的作用也引起了人们的注意。这项研究旨在检查RvD1对RA中pan的形成的作用及其潜在机制。方法采用UPLC-MS / MS和ELISA法分别测定RA患者和健康人的血清RvD1和CTGF水平。通过qRT-PCR和ELISA评估CTGF和炎性因子的水平。通过miRNA微阵列确定MicroRNA表达谱。CTGF，RvD1和miR-146a-5p对血管生成的影响通过试管形成和鸡绒膜尿囊膜（CAM）分析进行评估。构建胶原诱导的关节炎（CIA）小鼠，以检测RvD1和miR146a-5p对RA的作用。STAT3激活通过蛋白质印迹法确定。结果RA患者血清中RvD1水平降低而CTGF水平升高，同时观察到RA患者血清中RvD1和CTGF浓度呈负相关。在CIA小鼠中，RvD1抑制了血管生成并降低了CTGF的表达。同时，RvD1显着降低RA FLS中的CTGF和促炎细胞因子水平。此外，CTGF抑制血管生成，RvD1抑制RA FLS和血管生成的增殖和迁移。MiRNA芯片和qRT-PCR结果表明RvD1上调了miRNA-146a-5p。转染实验表明，miRNA-146a-5p可以降低炎症因子和CTGF水平。而且，miRNA-146a-5p降低了体内FLS的增殖和血管生成。MiRNA-146a-5p还可抑制CIA小鼠的血管生成并下调CTGF的表达。最后，蛋白质印迹结果显示miRNA-146a-5p抑制STAT3的激活。结论RvD1易于通过上调miRNA-146a-5p抑制RA的进展，从而抑制CTGF和炎症介质的表达，从而减少血管pan的形成和软骨损伤。MiRNA芯片和qRT-PCR结果表明RvD1上调了miRNA-146a-5p。转染实验表明，miRNA-146a-5p可以降低炎症因子和CTGF水平。而且，miRNA-146a-5p降低了体内FLS的增殖和血管生成。MiRNA-146a-5p还可抑制CIA小鼠的血管生成并下调CTGF的表达。最后，蛋白质印迹结果显示miRNA-146a-5p抑制STAT3的激活。结论RvD1易于通过上调miRNA-146a-5p抑制RA的进展，从而抑制CTGF和炎症介质的表达，从而减少血管pan的形成和软骨损伤。MiRNA芯片和qRT-PCR结果表明RvD1上调了miRNA-146a-5p。转染实验表明，miRNA-146a-5p可以降低炎症因子和CTGF水平。而且，miRNA-146a-5p降低了体内FLS的增殖和血管生成。MiRNA-146a-5p还可抑制CIA小鼠的血管生成并下调CTGF的表达。最后，蛋白质印迹结果显示miRNA-146a-5p抑制STAT3的激活。结论RvD1倾向于通过上调miRNA-146a-5p抑制RA的发展，从而抑制CTGF和炎症介质的表达，从而减少血管pan的形成和软骨损伤。miRNA-146a-5p降低了FLS的增殖和体内血管生成。MiRNA-146a-5p还可抑制CIA小鼠的血管生成并下调CTGF的表达。最后，蛋白质印迹结果显示miRNA-146a-5p抑制STAT3的激活。结论RvD1易于通过上调miRNA-146a-5p抑制RA的进展，从而抑制CTGF和炎症介质的表达，从而减少血管pan的形成和软骨损伤。miRNA-146a-5p降低了FLS的增殖和体内血管生成。MiRNA-146a-5p还可抑制CIA小鼠的血管生成并下调CTGF的表达。最后，蛋白质印迹结果显示miRNA-146a-5p抑制STAT3的激活。结论RvD1倾向于通过上调miRNA-146a-5p抑制RA的发展，从而抑制CTGF和炎症介质的表达，从而减少血管pan的形成和软骨损伤。