来源：风湿病与关节炎,2019,8（1）:10-14.

活血通络方对类风湿关节炎滑膜成纤维细胞miR-155及相关细胞因子表达的影响（基础研究）

靖卫霞¹，朱跃兰²，马俊福³，孙文燕⁴

　　【摘　要】目的：观察活血通络方对白细胞介素-1β（IL-1β）刺激类风湿关节炎（RA）滑膜成纤维细胞（HFLS）增殖和miR-155以及相关细胞因子分泌的影响。方法：HFLS-RA培养到第4代至第8代。经IL-1β诱导后给予含药血清，采用MTT法检测空白组、模型组、对照组、低剂量组、中剂量组、高剂量组含药血清对HFLS-RA增殖的影响，荧光定量PCR法检测miR-155及相关细胞因子表达。结果：MTT法测定HFLS-RA增殖活性的结果显示，模型组比正常组细胞数量明显增多（P < 0.01），对照组和高、中剂量组增殖活性明显降低（P < 0.01），低剂量组未表现出对HFLS-RA增殖有显著影响。对照组及低、中、高剂量组miR-155、IL-2、IFN-γ和RANKL相对表达水平均低于模型组，IL-4、IL-10相对表达水平高于模型组（P < 0.05）。高剂量组miR-155、IL-2、IFN-γ和RANKL相对表达水平均低于对照组，IL-4、IL-10相对表达水平高于对照组（P < 0.05），中剂量组miR-155、IL-2、IL-4、IL-10、IFN-γ和RANKL相对表达水平与对照组比较，差异无统计学意义（P > 0.05）。结论：活血通络方抑制HFLS-RA增殖，其治疗RA可能通过下调miR-155表达抑制RA滑膜成纤维细胞中IL-2、IFN-γ和RANKL等细胞因子表达，提高IL-4、IL-10等细胞因子表达相关。

　　【关键词】　关节炎，类风湿；活血通络方；miR-155；细胞因子；细胞实验；大鼠

Effect of Huoxue Tongluo Fang（活血通络方）on the Expression of miR-155 and Related Cytokines in Synovial Fibroblasts of Rheumatoid Arthritis

JING Wei-xia,ZHU Yue-lan,MA Jun-fu,SUN Wen-yan

　　【ABSTRACT】Objective:To observe the effects of Huoxue Tongluo Fang（活血通络方）on the proliferation of rheumatoid arthritis（RA）synovial fibroblasts（HFLS）stimulated by interleukin-1β（IL-1β）and the secretion of miR-155 and related cytokines.Methods:HFLS-RA was cultured to the fourth to eighth generations.After induction by IL-1β,drug-containing serum was given.MTT assay was used to detect the effect of blank group,model group,control group,low-dose group,medium-dose group and high-dose group of drug-containing serum on the proliferation of HFLS-RA.Fluorescence quantitative PCR was used to detect the expression of miR-155 and related cytokines.Results:MTT assay showed that the number of cells in the model group was significantly higher than that in the normal group（P < 0.01）.The proliferative activity of the control group and the high-dose and medium-dose groups was significantly lower（P < 0.01）.The low-dose group did not show significant effect on the proliferation of HFLS-RA.The relative expression levels of miR-155,IL-2,IFN-γ and RANKL in the control group,the low-dose group,the medium-dose group and the high-dose group were lower than those in the model group,while the relative expression levels of IL-4 and IL-10 were higher than those in the model group（P < 0.05）.The relative expression levels of miR-155,IL-2,IFN-γ and RANKL in the high dose group were lower than those in the control group,while the relative expression levels of IL-4 and IL-10 were higher than those in the control group（P < 0.05）.There was no significant difference in the relative expression levels of miR-155,IL-2,IL-4,IL-10,IFN-γ and RANKL between the middle dose group and the control group（P > 0.05）.Conclusion:Huoxue Tongluo Fang can inhibit the proliferation of HFLS-RA,and its treatment for RA may inhibit the expression of IL-2,IFN-γ and RANKL in RA synovial fibroblasts by down-regulating the expression of miR-155,and increase the expression of IL-4,IL-10 and other cytokines.

　　【Keywords】 arthritis,rheumatoid;Huoxue Tongluo Fang（活血通络方）;miR-155;cytokines;cell experi-ment;rats