透骨血竭散对类风湿关节炎滑膜成纤维细胞的作用机制  

Effect of Tougu Xuejie Powder on Synovial Fibroblasts in Rat Model of Rheumatoid Arthritis

作　　者：邓文雯 肖智 姜如 DENG Wen-wen;XIAO Zhi;JIANG Ru(Department of Integrative Traditional Chinese and Western Medicine,the Third Hospital of Changsha,Changsha Hunan 410035)

机构地区：[1]长沙市第三医院中西医结合科,湖南长沙410035

出　　处：《世界中西医结合杂志》2023年第12期2379-2385,共7页World Journal of Integrated Traditional and Western Medicine

基　　金：长沙市自然科学基金项目(kq2014011)。

摘　　要：目的探讨透骨血竭散(TGXJS)对类风湿关节炎(Rheumatoid arthritis,RA)大鼠滑膜成纤维细胞(Synovial fibroblasts,SFs)增殖、凋亡和炎症因子的影响及可能的作用机制。方法培养大鼠SFs,并设置细胞分组,细胞分为5组,分别为对照组(正常培养不做处理,Control),模型组(IL-1β处理24 h,Model),透骨血竭散组(IL-1β和透骨血竭散共处理24 h,TGXJS)、透骨血竭散+sh-ALX3组(转染ALX3基因敲降质粒24 h后,加IL-1β和透骨血竭散共处理24 h,TGXJS+sh-ALX3),透骨血竭散+sh-ALX3+OE-ITGB1组(转染ALX3基因敲降质粒和ITGB1基因过表达质粒24 h后,加IL-1β和透骨血竭散共处理24 h,TGXJS+sh-ALX3+OE-ITGB1)。Western-blot检测各组细胞ALX3和ITGB1蛋白表达情况;CCK8细胞增殖实验检测各组细胞72 h后细胞增殖率;ELISA法检测各组细胞炎症因子IL-6、IL-10和TNF-α水平;流式细胞术检测各组细胞的凋亡情况。结果与对照组比较,模型组ALX3和ITGB1蛋白表达降低,细胞增殖率增高,IL-6、IL-10和TNF-α水平提升,细胞凋亡率降低,差异有统计学意义(P<0.05)。与模型组比较,透骨血竭散组ALX3和ITGB1蛋白表达增高,细胞增殖率降低、IL-6和TNF-α水平降低,IL-10水平增加,细胞凋亡率增加,差异有统计学意义(P<0.05);与透骨血竭散组比较,透骨血竭散+sh-ALX3组,ALX3和ITGB1蛋白表达降低,细胞增殖率增高,IL-6和TNF-α水平水平上升,IL-10水平下降,细胞凋亡率降低,差异有统计学意义(P<0.05),透骨血竭散+sh-ALX3+OE-ITGB1组ALX3蛋白表达降低,ITGB1蛋白表达上升,细胞增殖率降低,IL-6和TNF-α水平降低,IL-10水平上升,细胞凋亡增加,差异有统计学意义(P<0.05)。结论透骨血竭散通过激活ALX3基因,上调ITGB1表达,抑制RA大鼠SFs增殖、凋亡和炎症反应。Objective To investigate the effects of Tougu Xuejie Powder(TGXJS)on the proliferation,apoptosis,and inflammatory cytokines of synovial fibroblasts(SFs)in the rat model of rheumatoid arthritis(RA)and decipher the possible mechanism.Methods Rat SFs were cultured and classified into the control(normal culture without treatment),model[interleukin(IL)-1βtreatment for 24 h],TGXJS(IL-1β+TGXJS treatment for 24 h),TGXJS+sh-ALX3[24 h after transfection with the plasmid for the knockdown of the aristaless-like homeobox 3(ALX3)gene,cells were treated with IL-1β+TGXJS for 24 h],and TGXJS+sh-ALX3+OE-ITGB1[24 h after transfection with the plasmid for the knockdown of ALX3 gene and the plasmid for the overexpression of the integrinβ1(ITGB1)gene,cells were treated with IL-1β+TGXJS for 24 h]groups.Western blotting was employed to determine the protein levels of ALX3 and ITGB1 in each group.The CCK8 assay kit was used to detect the cell proliferation rate of each group after 72 h.The levels of inflammatory cytokines[IL-6,IL-10,and tumor necrosis factor(TNF)-α]were measured by the enzyme-linked immunosorbent assay.The apoptosis of cells in each group was detected by flow cytometry.Results Compared with the control group,the model group showed down-regulated expression of ALX3 and ITGB1,increased cell proliferation rate,elevated levels of IL-6,IL-10,and TNF-α,and decreased cell apoptosis rate(P<0.05).Compared with the model group,the TGXJS group presented up-regulated protein levels of ALX3 and ITGB1,decreased cell proliferation rate,lowered IL-6 and TNF-αlevels,elevated IL-10 level,and increased apoptosis rate(P<0.05).Compared with the TGXJS group,the TGXJS+sh-ALX3 group showed down-regulated protein levels of ALX3 and ITGB1,increased cell proliferation rate,elevated IL-6 and TNF-αlevels,lowered IL-10 level,and decreased apoptosis rate(P<0.05).The TGXJS+sh-ALX3+OE-ITGB1 group with down-regulated protein level of ALX3 and up-regulated protein level of ITGB1 showed decreased cell proliferation rate,lowered IL-6 and TNF-αlev

关 键 词：透骨血竭散 转录调节因子Alx3 β1型整合素 滑膜成纤维细胞 增殖 凋亡 炎症反应 

分 类 号：R285.5[医药卫生—中药学]