干扰NLRP3及NLRP12对IL-6诱导全身型幼年特发性关节炎滑膜成纤维细胞增殖影响及机制研究  

Effects and Mechanisms of Interference with NLRP3 and NLRP12 on IL-6 Induced Synovial Fibroblast Proliferation in Systemic Juvenile Idiopathic Arthritis

作　　者：谢颖[1] 李丰[1] 唐盈 程苏云 张松 韦茹 徐彩虹 曾华松[1] XIE Ying;LI Feng;TANG Ying;CHENG Su-yun;ZHANG Song;WEI Ru;XU Cai-hong;ZENG Hua-song(Departmentof Rheumatology and Immunology of Guangzhou Women and Children's Medical Center Affiliated to Guangzhou Medical University,510000)

机构地区：[1]广州医科大学附属广州市妇女儿童医疗中心风湿免疫科,510120

出　　处：《岭南急诊医学杂志》2023年第5期401-403,412,共4页Lingnan Journal of Emergency Medicine

基　　金：广东省自然科学基金(2017A030313557);广东省广州市卫生局课题(20121A011071)。

摘　　要：目的:全身型幼年特发性关节炎(systemic juvenile idiopathic arthritis,sJIA)目前免疫机制尚不明确。本研究旨在探讨炎症小体对白介素(IL)6诱导的类风湿关节炎滑膜成纤维(Rheumatoid Arthritis Synovial Fibroblasts,RASF)细胞增殖及炎症因子分泌的影响。方法:构建IL-6诱导的RASF细胞增殖模型和设计合成2条核苷酸结合寡聚结构域样受体蛋白(NOD-like receptor protein)3和NLRP12的siRNA干扰片段,通过脂质体转染RASF细胞,建立NLRP3,NLRP12干扰的细胞株;实时荧光定量PCR和ELISA检测NLRP3、NLRP12 mRNA及炎症因子IL-1,IL-18,肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)的表达;再用IL-6处理细胞,MMT法检测细胞增殖;qPCR检测NLRP通路相关分子及核因子κB(NF-κB)p65,核因子κB受体活化因子配体(receptor activator of NF-κB ligand,RANKL)表达的影响。结果:IL-6呈剂量依赖性诱导的RASF细胞增殖增殖,NLRP3 mRNA和蛋白表达水平均明显升高(P<0.05),NLRP12较对照组无明显变化(P>0.05);转染siRNA后可显著降低NLRP3,NLRP12在RASF细胞内mRNA的表达(P<0.05),且NLRP3表达降低后会抑制IL-6诱导的RASF细胞增殖,而抑制NLRP12则促进IL-6诱导的RASF细胞增殖。结论:NLRP3与促进IL-6诱导的RASF细胞增殖有关,NLRP12可能参与抑制RASF细胞增殖。Objective:The mechanism of systemic juvenile idiopathic arthritis(sJIA)is still unknown.Objective to investigate the effect of inflammasome on IL-6-induced proliferation of RASF cells and secretion of inflammatory factors.Methods:The proliferation model of RASF cells induced by IL-6 was constructed;two siRNA interference fragments of NLRP3 and nlrp12 were designed and synthesized,and then transfected into RASF cells by liposome to establish NLRP3 and nlrp12 interference cell lines;real-time fluorescent quantitative PCR and ELISA were used to detect the expression of NLRP3,nlrp12 mRNA and inflammatory factors IL-1,IL-18 and TNF-a;Then IL-6 was used to treat the cells,and MMT was used to detect cell proliferation;qPCR was used to detect nlrp pathway related molecules and p65,RANKL expression.The mRNA expressions of NLRP3,NLRP12 and pathway related molecules were measured by qPCR.Results:the proliferation of RASF cells was induced by IL-6 in a dose-dependent manner,and the expression levels of NLRP3 mRNA and protein were significantly increased(P<0.05),but there was no significant change in nlrp12 compared with the control group(P>0.05);After transfection with siRNA,the expression of NLRP3 and nlrp12 mRNA in RASF cells was significantly decreased(P<0.05).The decreased expression of NLRP3 inhibited the proliferation of RASF cells induced by IL-6,while the inhibition of nlrp12 promoted the proliferation of RASF cells induced by IL-6.Conclusion:NLRP3 may be involved in promoting the proliferation of RASF cells induced by IL-6,and nlrp12 may be involved in inhibiting the proliferation of RASF cells.

关 键 词：炎症小体 干扰RNA 全身型幼年特发性关节炎 

分 类 号：R725.8[医药卫生—儿科]