Resolvin D1 suppresses pannus formation via decreasing connective tissue growth factor caused by upregulation of miRNA-146a-5p in rheumatoid arthritis
Resolvin D1 suppresses pannus formation via decreasing connective tissue growth factor caused by upregulation of miRNA-146a-5p in rheumatoid arthritis
复制标题
Resolvin D1 通过减少类风湿性关节炎中 miRNA-146a-5p 上调引起的结缔组织生长因子来抑制血管翳形成
DOI：
10.1186/s13075-020-2133-2
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发表时间：
2020-03-27
影响因子：
4.9
通讯作者：
Wang, Jianguang
中科院分区：
文献类型：
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作者：
Sun, Weiwei;Ma, Jinglan;Wang, Jianguang
背景类风湿性关节炎（RA）是一种慢性自身免疫性疾病，其特征是炎症和关节僵硬，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，可促进成纤维细胞样滑膜细胞 (FLS) 增殖、血管翳形成以及软骨和骨损伤。 Resolvin D1（RvD1）可以促进急性炎症性疾病的炎症消退，最近，RvD1对慢性炎症性疾病的作用也引起了人们的关注。本研究旨在探讨RvD1对RA血管翳形成的影响及其机制。方法分别采用UPLC-MS/MS和ELISA法测定RA患者和健康人血清中RvD1和CTGF的水平。通过 qRT-PCR 和 ELISA 评估 CTGF 和炎症因子的水平。 MicroRNA 表达谱通过 miRNA 微阵列测定。通过管形成和鸡绒毛尿囊膜 (CAM) 测定评估 CTGF、RvD1 和 miR-146a-5p 对血管生成的影响。构建胶原诱导关节炎（CIA）小鼠来检测RvD1和miR146a-5p对RA的影响。 Western blotting检测STAT3活化情况。结果RA患者血清中RvD1水平降低，CTGF水平升高，且同时观察到RA患者血清中RvD1和CTGF浓度呈负相关。在 CIA 小鼠中，RvD1 抑制血管生成并降低 CTGF 的表达。同时，RvD1 显着降低 RA FLS 中的 CTGF 和促炎症细胞因子水平。此外，CTGF 抑制血管生成，RvD1 抑制 RA FLS 的增殖和迁移以及血管生成。 miRNA 微阵列和 qRT-PCR 结果显示 RvD1 上调 miRNA-146a-5p。转染实验表明，miRNA-146a-5p 可以降低炎症因子和 CTGF 水平。此外，miRNA-146a-5p 降低了体内 FLS 的增殖和血管生成。 miRNA-146a-5p 还抑制 CIA 小鼠的血管生成并下调 CTGF 的表达。最后，Western blot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CTGF和炎症介质的表达来减轻RA进展，从而减少血管翳形成和软骨损伤。
BackgroundRheumatoid 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.MethodsSerum 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.ResultsRvD1 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.ConclusionRvD1 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.