C反应蛋白在类风湿关节炎发病中的作用机制

发布时间：2018-09-10 14:24

【摘要】：目的了解C反应蛋白在类风湿关节炎发病机制中的作用。方法体外培养RA及正常滑膜成纤维细胞,qRT-PCR检测RA及正常滑膜成纤维细胞中CRP及其受体CD32、CD64 mRNA的表达,免疫组化及western blot检测RA及正常滑膜组织中CRP、CD32、CD64蛋白的表达。加入外源性CRP刺激RA及正常滑膜成纤维细胞后检测各种炎症因子和金属基质蛋白酶MMPs的jmRNA表达及RA滑膜成纤维细胞磷酸化p65的表达。分别加入CD32拮抗剂和NF-κB拮抗剂后再重复CRP刺激检测RA滑膜成纤维细胞炎症因子mRNA的表达。同样,分别加入IL-1β、 TNF-α刺激RA滑膜成纤维细胞检测CRP mRNA的表达。加入NF-κB拮抗剂后再重复IL-1β、TNF-α刺激RA滑膜成纤维细胞检测CRP mRNA的表达。结果与正常滑膜成纤维细胞相比,RA中CRP及其受体CD32和CD64mRNA及蛋白表达均明显升高。加入外源性CRP刺激后RA滑膜成纤维细胞中各种炎症因子和MMP-3较刺激前升高,RA滑膜成纤维细胞p65的磷酸化水平升高。加入CD32拮抗剂再重复CRP刺激后RA滑膜成纤维细胞中各炎症因子和MMP-3mRNA表达比加入拮抗剂前的作用明显减弱,加入NF-κB拮抗剂再重复CRP刺激后RA滑膜成纤维细胞中各炎症因子和MMP-3 mRNA表达比加入拮抗剂前的作用明显减弱。IL-1β或TNF-α刺激RA滑膜成纤维细胞检测CRP mRNA的表达明显升高,加入NF-κB拮抗剂后再重复IL-1β或TNF-α刺激则CRP mRNA表达明显减弱。结论在RA中CRP通过与受体CD32、CD64结合,激活NF-κB通路,刺激各种炎症因子和MMP-3的分泌。同时IL-1 β和TNF-α的产生又可通过激活NF-κB通路,刺激CRP的产生,周而复始,导致炎症反复加重,病情迁延不愈。CRP在RA中不仅是标志物,而且参与其发病。
[Abstract]:Objective to investigate the role of C reactive protein in the pathogenesis of rheumatoid arthritis. Methods the expression of CRP and its receptor CD32,CD64 mRNA in RA and normal synovial fibroblasts was detected by qRT-PCR in cultured RA and normal synovial fibroblasts, and the expression of CRP,CD32,CD64 protein in RA and normal synovial tissues was detected by immunohistochemistry and western blot. After RA and normal synovial fibroblasts were stimulated by exogenous CRP, the jmRNA expression of various inflammatory factors and metalloproteinases MMPs and the expression of phosphorylated p65 in RA synovial fibroblasts were detected. After adding CD32 antagonist and NF- 魏 B antagonist, the expression of inflammatory factor mRNA in RA synovial fibroblasts was detected by repeated CRP stimulation. Similarly, RA synovial fibroblasts were stimulated by IL-1 尾 and TNF- 伪 to detect the expression of CRP mRNA. After adding NF- 魏 B antagonist, IL-1 尾 -TNF- 伪 stimulated RA synovial fibroblasts to detect the expression of CRP mRNA. Results compared with normal synovial fibroblasts, the expression of CRP and its receptors, CD32, CD64mRNA and protein in RA were significantly increased. The levels of p65 phosphorylation of RA synovial fibroblasts were increased by adding exogenous CRP to RA synovial fibroblasts after stimulation of various inflammatory factors and MMP-3. The expression of inflammatory factors and MMP-3mRNA in RA synovial fibroblasts after repeated CRP stimulation with CD32 antagonist was significantly weaker than that before adding the antagonist. After repeated CRP stimulation with NF- 魏 B antagonist, the expression of inflammatory factors and MMP-3 mRNA in RA synovial fibroblasts was significantly decreased. IL-1 尾 or TNF- 伪 stimulated RA synovial fibroblasts significantly increased CRP mRNA expression. After the addition of NF- 魏 B antagonist and repeated stimulation of IL-1 尾 or TNF- 伪, the expression of CRP mRNA decreased significantly. Conclusion CRP activates the NF- 魏 B pathway and stimulates the secretion of various inflammatory factors and MMP-3 by binding to the receptor CD32,CD64 in RA. At the same time, the production of IL-1 尾 and TNF- 伪 can stimulate the production of CRP by activating the NF- 魏 B pathway, which leads to the repeated exacerbation of inflammation, and it is not only a marker in RA, but also involved in the pathogenesis of RA.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.22

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