雷公藤红素抑制TNF-α诱导的类风湿关节炎作用及其机制研究  

Study on the Inhibitory Effect and Mechanism of Celastrol Against Rheumatoid Arthritis Induced by TNF-α

作　　者：李科[1] 曹玉净[1,2] 钱亚男 李光辉 周松林[1] Li Ke;Cao Yujing;Qian Yanan;Li Guanghui;Zhou Songlin(Henan Provincial Hospital of Traditional Chinese Medicine,Zhengzhou 450053,China;College ofOrthopedics and Traumatology,Henan University of Chinese Medicine,Zhengzhou 450002,China)

机构地区：[1]河南省中医院,河南郑州450053 [2]河南中医药大学骨伤学院,河南郑州450002

出　　处：《亚太传统医药》2025年第3期16-21,共6页Asia-Pacific Traditional Medicine

基　　金：国家自然科学基金项目(82104833);河南省中医药科学研究专项课题(2024ZYZD06,2023ZY1008,2019ZYBJ15)。

摘　　要：目的:探究雷公藤红素抑制TNF-α诱导的类风湿关炎的作用及机制。方法:采用类风湿性关节炎成纤维细胞MH7A建立体外类风湿关节炎模型,并将实验分为空白组、模型组(TNF-α10 ng/mL)和给药组。以10%胎牛血清和1%青霉素-链霉素混合溶液的DMEM高糖培养基处理的MH7A细胞作为对照组,在MH7A细胞中加入浓度为10 ng/mL的TNF-α处理24 h作为模型组,在MH7A细胞中加入系列浓度的雷公藤红素处理2 h后再加入浓度为10 ng/mL的TNF-α共处理24 h作为给药组。使用CCK-8法检测雷公藤红素对MH7A的细胞毒性,并检测雷公藤红素对TNF-α诱导的RA-FLS滑膜炎症细胞异常增殖的抑制活性,利用酶联免疫吸附测定法(Elisa法)测定雷公藤红素对RA-FLS产生的IL-1β和IL-6和基质金属蛋白酶(MMP-1和MMP-3)表达的影响;Western blot检测雷公藤红素对RA-FLS凋亡相关蛋白和机制相关蛋白表达的影响。结果:与空白组相比,模型组被TNF-α激活,炎性细胞因子(P<0.01)、基质金属蛋白酶均显著性地增加(P<0.0001);与模型组比较,给药组的RA-FLS的增殖被显著性地抑制(P<0.001),IL-1β、IL-6、MMP-1及MMP-3的表达也显著性地降低;雷公藤红素治疗后能抑制磷酸化ERK通路蛋白的激活(P<0.01),促进RA-FLS的凋亡(P<0.05)。结论:雷公藤红素抑制TNF-α诱导的类风湿关节炎的作用机制可能与其抑制磷酸化ERK相关通路蛋白的激活,促进RA-FLS的凋亡相关。Objective:To explore the effect and mechanism of Celastrol in inhibiting tumor necrosis factor-alpha(TNF-α)induced rheumatoid arthritis.Methods:An in vitro rheumatoid arthritis model was established using human rheumatoid arthritis fibroblast-like synoviocytes(RA-FLS)MH7A.The groups were divided into a blank group,a model group(TNF-α 10 ng/mL),and a treatment group.MH7A cells treated with high-glucose DMEM culture medium containing 10% fetal bovine serum and 1% penicillin-streptomycin mixture served as the control group.MH7A cells were treated with a final concentration of 10 ng/mL TNF-α for 24 hours as the model group.MH7A cells were treated with a series of concentrations of Celastrol for 2 hours followed by a final concentration of 10 ng/mL TNF-α for 24 hours as the treatment group.The cytotoxicity of Celastrol on MH7A was detected using the CCK-8 assay,and the inhibitory activity of Celastrol on the abnormal proliferation of TNF-αinduced RA-FLS synovial inflammatory cells was detected.The expression of interleukin-1 beta(IL-1β)and interleukin-6(IL-6)and matrix metalloproteinases(MMP-1 and MMP-3)produced by RA-FLS was measured using the enzyme-linked immunosorbent assay(ELISA).The expression of apoptosis-related proteins and mechanism-related proteins in RA-FLS was detected by Western blot.Results:Compared to the control group,the model group showed a significant increase in inflammatory cytokines(P<0.01)and matrix metalloproteinases after being activated by TNF-α(P<0.0001).The proliferation of RA-FLS in the drug groups was significantly inhibited(P<0.001),and the expression of IL-1β,IL-6,MMP-1,and MMP-3 was also significantly reduced compared to the model group.Celastrol treatment inhibited the activation of phosphorylated ERK pathway proteins(P<0.01)and promoted apoptosis in RA-FLS(P<0.05).Conclusion:The mechanism by which Celastrol inhibits tumor necrosis factor-alpha(TNF-α)induced rheumatoid arthritis may be related to its inhibition of the activation of phosphorylated extracellular signal-regulat

关 键 词：类风湿关节炎 类风湿关节炎成纤维样滑膜细胞 雷公藤红素 肿瘤坏死因子Α 凋亡 ERK 

分 类 号：R269[医药卫生—中西医结合]