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| 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 |
| Wogonin induces apoptosis of fibroblast-like synoviocytes via reactive oxygen species-dependent activation of p38 mitogen-activated protein kinase in rheumatoid arthritis |
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| DOI：10.3969/j.issn.1006.7108.2017.07.011 |
| 中文关键词: 汉黄芩素 类风湿关节炎 成纤维样滑膜细胞 细胞凋亡 中医中药 |
| 英文关键词:Wogonin Rheumatoid arthritis Fibroblast like synoviocytes Cell apoptosis Traditional Chinese Medicine |
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| 中文摘要: |
| 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)调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20 (μg/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。 |
| 英文摘要: |
| Objective To explore the effect of wogonin on the apoptosis of rheumatoid arthritis fibroblast-like synoviocytes (RA- FLS) and the associated molecular mechanisms. Methods FLSs were harvested from RA patients and cultured in joint synovial fluid and through 3-5 passages. With different treatments, cells were divided into six groups： control group, low-dose wogonin group (20μg /mL of wogonin) , middle-dose of wogonin group (50μg /mL of wogonin)，high-dose of wogonin group (100μg / mL of wogonin) , 100μg /mL Wogonin + NAC group, and 100μg /mL Wogonin + SB203580 group. The proliferation of RA-FLS was measured with MTT assay. The apoptosis was measured with flow cytometric analysis. Mitochondrial membrane potential (MMP) was measured with rhodamine 123 staining. The intracellular levels of reactive oxygen species (ROS) were tested with DCFH-DA staining. The expression of proteins in p38 MAPK pathway was examined with Western blotting. Results Compared to the control group, different concentrations of wogonin inhibited RA-FLS growth, induced apoptosis, decreased mitochondrial membrane potential, and promoted p38MAPK phosphorylation and ROS production, and was in a concentration-dependent pattern. The apoptotic ratio of wogonin-treated group was markedly decreased by the ROS scavenger NAC or the p38MAPK inhibitor SB203580. and pretreatment of RA-FLS with 5 mmol/L NAC markedly reversed the wogonin-induced enhancement of p-p38 MAPK. Conclusion Wogonin effectively induces the apoptosis of FLS from rheumatoid arthritis patients through the induction of ROS formation and p38 MAPK activation. |
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