大黄素对人类风湿关节炎成纤维样滑膜细胞自噬的影响
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湖北民族大学风湿性疾病发生与干预湖北省重点实验室

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国家自然科学基金项目(81860757):三百棒有效成分群通过PI3K/Akt/mTOR信号通路调控RA成纤维细胞自噬与滑膜增殖的作用机制


Effects of emodin on autophagy and cycle arrest in fibroblast-like synoviocytes in rheumatoid arthritis
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Hubei Minzu university Hubei Key Laboratory of Occurrence Intervention of Rheumatic Diseases

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    目的:基于cGAS/STING信号通路研究大黄素(EMD)对人类风湿关节炎成纤维样滑膜细胞(MH7A)细胞自噬的潜在作用。方法:采用CCK-8法检测MH7A细胞增殖的结果,并根据细胞存活率筛选出药物的浓度,并加入自噬抑制剂3-MA进一步验证大黄素对自噬的影响;采用MDC法检测MH7A细胞自噬情况;采用蛋白免疫印迹法(Western blot)检测cGAS、STING、p-STING、LC3-I、LC3-II、p62、Beclin-1的蛋白表达水平。结果:MDC染色结果提示,大黄素能够增强MH7A细胞自噬;Western Blot结果提示,大黄素可降低MH7A细胞自噬相关蛋白cGAS、STING、p-STING、P62的表达,增加LC3-II和Beclin-1的表达。加入自噬抑制剂3-MA后,MH7A细胞P62蛋白表达升高,LC3-II和Beclin-1蛋白表达降低。结论:大黄素可能通过下调cGAS/STING信号通路加速自噬,抑制MH7A细胞增殖。

    Abstract:

    Objective: Based on the cGAS/STING signaling pathway to investigate the potential effect of emodin (EMD) on autophagy of human rheumatoid arthritis fibroblast synovial cells (MH7A). Methods: CCK-8 method was used to detect the results of MH7A cell proliferation, and the concentration of drug was screened according to cell survival rate. Then, autophagy inhibitor 3-MA was added to further verify the effect of emodin on autophagy. Autophagy of MH7A cells was detected by MDC method. The protein expression levels of cGAS, STING, p-STING, LC3-I, LC3-II, p62 and Beclin-1 were detected by Western blot. Results: MDC staining indicated that emodin could enhance autophagy of MH7A cells. Western Blot results indicated that emodin could decrease the expressions of autophagy related proteins cGAS, STING, p-STING and P62, and increase the expressions of LC3-II and Beclin-1 in MH7A cells. After addition of autophagy inhibitor 3-MA, the expression of P62 protein in MH7A cells increased, while the expression of LC3-II and Beclin-1 protein decreased. Conclusion: Emodin may accelerate autophagy and inhibit MH7A cell proliferation by down-regulating cGAS/STING signaling pathway.

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  • 收稿日期:2023-04-09
  • 最后修改日期:2023-09-13
  • 录用日期:2024-01-05
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