汉黄芩素通过激活活性氧簇介导的p38MAPK信号通路诱导类风湿关节炎成纤维样滑膜细胞凋亡

发布时间：2018-09-15 19:19

【摘要】：目的探讨汉黄芩素对类风湿关节炎成纤维样滑膜细胞(rheumatoid arthritis fibroblast-like synoviocytes,RA-FLS)凋亡的影响以及作用机制。方法类风湿关节炎患者关节滑液经原代培养,应用3~5代传代的成纤维样滑膜细胞。按照不同方法处理分为6组:空白对照组、低剂量汉黄芩素组(终浓度为20μg/mL)、中剂量汉黄芩素组(终浓度为50μg/mL)、高剂量汉黄芩素组(终浓度为100μg/mL)、100μg/mL汉黄芩素+NAC组、100μg/mL汉黄芩素+SB203580组。MTT法检测细胞增殖情况,流式细胞仪检测细胞凋亡率,罗丹明123染色荧光显微镜照相法检测线粒体膜电位(mitochondrial membrane potential,MMP),DCFH-DA染色荧光显微镜照相法检测细胞内活性氧簇(reactive oxygen species,ROS)水平,Western blot法检测p38MAPK通路相关蛋白的表达。结果与正常对照组相比,各剂量汉黄芩素均能够抑制RA-FLS细胞的增殖,增加其凋亡,降低MMP以及增加ROS水平,激活p38MAPK信号通路,并且呈现剂量依赖性关系。ROS清除剂NAC和p38抑制剂SB203580可以明显降低汉黄芩素诱导的RA-FLS细胞的凋亡,并且5 mmol/L NAC下调汉黄芩素引起的p38MAPK磷酸化。结论汉黄芩素能够诱导RA-FLS细胞凋亡,ROS/p38MAPK信号通路参与了其凋亡的过程。
[Abstract]:Objective to investigate the effect of baicalin on apoptosis of fibroid synovial cells (rheumatoid arthritis fibroblast-like synoviocytes,RA-FLS) in rheumatoid arthritis (RA) and its mechanism. Methods the synovial fluid of patients with rheumatoid arthritis was cultured in primary culture. According to different methods, they were divided into 6 groups: blank control group, The cell proliferation was detected with low dose of baicalin (20 渭 g/mL), middle dose (50 渭 g/mL) and high dose (final concentration of 100 渭 g/mL) of 100 渭 g/mL and 100 渭 g/mL SB203580. The apoptosis rate was detected by flow cytometry. Detection of mitochondrial membrane potential (mitochondrial membrane potential,MMP) by Rhodamine 123 staining fluorescence microscope) DCFH-DA staining fluorescence microscopy was used to detect the level of reactive oxygen species (reactive oxygen species,ROS) in cells. Western blot was used to detect the expression of p38MAPK pathway related proteins. Results compared with the control group, baicalin could inhibit the proliferation of RA-FLS cells, increase its apoptosis, decrease the level of MMP and increase the level of ROS, and activate the p38MAPK signaling pathway. In a dose-dependent manner, Ros scavenger NAC and p38 inhibitor SB203580 could significantly reduce the apoptosis of RA-FLS cells induced by baicalin, and 5 mmol/L NAC down-regulated p38MAPK phosphorylation induced by baicalin. Conclusion: baicalin can induce apoptosis of RA-FLS cells via Ros / p38 MAPK signaling pathway.
【作者单位】：河南省中医院风湿病科;河南中医药大学第一附属医院风湿病科;
【分类号】：R259

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