类叶牡丹提取物通过NF-κB信号通路影响类风湿关节炎成纤维样滑膜细胞的增殖及凋亡  

Effects of Caulophyllum robustum Maxim extract on the proliferation and apoptosis of fibroblast-like synoviocytes in rheumatoid arthritis through NF-κB signaling pathway

作　　者：曾茂湘 吕兆宇 李妍 ZENG Maoxiang;LYU Zhaoyu;LI Yan(Department of Surgery,the Second People′s Hospital of Longgang District,Shenzhen 518112,China)

机构地区：[1]深圳市龙岗区第二人民医院外二科,广东深圳518112

出　　处：《沈阳医学院学报》2020年第3期216-219,223,共5页Journal of Shenyang Medical College

基　　金：深圳市龙岗区医疗卫生科技计划项目(No.LGKCYLWS2019000048)

摘　　要：目的:研究类叶牡丹提取物(Caulophyllum robustum Maxim extract,CRME)对类风湿关节炎(rheumatoid arthritis,RA)成纤维样滑膜细胞(fibroblast-like synoviocytes,FLS)的增殖和凋亡的影响及相关分子机制。方法:取RA患者和非RA患者的滑膜组织,分离培养FLS,采用MTT法检测50、100、500μg/ml CRME和NF-κB信号通路抑制剂(PDTC)对FLS细胞活力的影响;采用ELISA法检测各剂量CRME对各组细胞白细胞介素-6(IL-6)、白细胞介素-4(IL-4)、血管内皮生长因子(VEGF)、Bax、Bcl-2和p65表达水平的影响;采用Annexin V-FITC流式双染检测CRME、PDTC对FLS细胞凋亡的影响。结果:RA-FLS组细胞增殖活性较正常组升高(P<0.05),RA-FLS+PDTC组和RA-FLS+各剂量CRME组RA-FLS细胞增殖活性较RA-FLS组降低(P<0.05)。和正常组比较,RA-FLS组细胞的IL-6、IL-4、VEGF、Bcl-2和p65表达水平升高(P<0.05);和RA-FLS组比较,RA-FLS+PDTC组和RA-FLS+50、100、500μg/ml CRME组RA-FLS细胞的IL-6表达水平降低,Bax表达水平升高(P<0.05),RA-FLS+PDTC组和RA-FLS+100、500μg/ml CRME组RAFLS细胞的IL-4表达水平升高(P<0.05),RA-FLS+500μg/ml CRME组RA-FLS细胞的VEGF表达水平降低(P<0.05),RA-FLS+PDTC组和RA-FLS+500μg/ml RA-FLS细胞的p65表达水平降低(P<0.05)。和RA-FLS组比较,RA-FLS+PDTC组和RA-FLS+100、500μg/ml CRME组RA-FLS细胞凋亡率均升高(P<0.05)。结论:CRME可降低VEGF水平,进而抑制血管翳形成,通过抑制NF-κB信号通路,减轻炎症反应,促进FLS凋亡,进而发挥抗RA的药理作用。Objective:To investigate the effects of Caulophyllum robustum Maxim extract(CRME)on the proliferation and apoptosis of fibroblast-like synoviocytes(FLS)in rheumatoid arthritis(RA)and the related molecular mechanism.Methods:Synovium tissues was taken from patients with RA and non-RA.FLS were prepared from synovium tissues.The effects of 50,100,500μg/ml CRME and PDTC(NF-κB pathway inhibitor)on the viability of FLS were detected by MTT method.The effects of different doses of CRME on the expression of IL-6,IL-4,VEGF,Bax,Bcl-2 and p65 were detected by ELISA method.The effects of CRME and PDTC on apoptosis of FLS were detected by Annexin V-FITC flow cytometry.Results:The proliferation activity of FLS in RA-FLS group was significantly higher than that in non-RA-FLS group(P<0.05),and the proliferation activity of RAFLS in PDTC group and CRME group was lower than that in RA-FLS group(P<0.05).Compared with the non-RA-FLS group,the expression levels of IL-6,IL-4,VEGF,Bcl-2 and p65 in RA-FLS group increased markedly(P<0.05).Compared with RA-FLS group,the IL-6 level decreased and the Bax level increased in PDTC group and 50,100,500μg/ml CRME groups(P<0.05),the IL-4 level increased in PDTC group and 100,500μg/ml CRME groups(P<0.05),the VEGF level decreased in the500μg/ml CRME group(P<0.05),and the p65 level decreased in PDTC group and 500μg/ml CRME group(P<0.05).Compared with RA-FLS group,apoptosis rate of RA-FLS cells increased in the PDTC group and 100,500μg/ml CRME groups(P<0.05).Conclusion:CRME can inhibit NF-κB signal pathway,alleviate inflammatory reaction,promote apoptosis of RA-FLS,inhibit pannus formation,and play the pharmacological role of anti-RA.

关 键 词：类叶牡丹提取物 类风湿关节炎 成纤维样滑膜细胞 细胞增殖 细胞凋亡 NF-ΚB信号通路 

分 类 号：R593[医药卫生—内科学]