昆母汤对类风湿关节炎成纤维样滑膜细胞bax、bcl-2的影响  

Effects of Kunmu Decoction on Bax and Bcl-2 Expression in Fibroblast-like Synoviocytes of Rheumatoid Arthritis Patients

作　　者：林昌松[1] 陈秀敏[2,3] 刘清平[1] 徐强[1] 关彤[1] 陈纪藩[1] 刘风震[4] 吴莹[4] 

机构地区：[1]广州中医药大学第一附属医院风湿病科,广东广州510405 [2]广东省中医院 [3]广州中医药大学博士后科研流动站,广东广州510120 [4]广州中医药大学,广东广州510405

出　　处：《中药新药与临床药理》2014年第6期656-659,共4页Traditional Chinese Drug Research and Clinical Pharmacology

基　　金：国家自然科学基金面上项目(81173634);广东省自然科学基金自由申请项目(S2011010005660)

摘　　要：目的观察昆母汤醇提液(KMT)对人类风湿关节炎成纤维样滑膜细胞(RA-FLS)B淋巴细胞瘤-2基因(bcl-2)、bcl-2相关X蛋白(bax)m RNA及蛋白的影响,探讨昆母汤治疗RA的部分作用机制。方法滑膜组织取自行关节置换术或关节镜检查术的8例活动性类风湿关节炎(RA)患者,分离出FLS并进行原代培养,用逆转录-聚合酶链反应(RT-PCR)观察bax、bcl-2 m RNA表达的变化,采用酶联免疫吸附测定(ELISA)法检测bax、bcl-2蛋白含量的变化。结果与肿瘤坏死因子-α(TNF-α)组比较,KMT高剂量组可以明显促进bax m RNA表达,明显抑制bcl-2 m RNA表达,且bax m RNA/bcl-2 m RNA比值明显提高(P<0.05,P<0.01);KMT中、高剂量组可以提高bax蛋白含量,各剂量组均可降低bcl-2蛋白含量,bax/bcl-2的蛋白比值明显提高(P<0.05,P<0.01),且有一定的剂量依赖性。结论昆母汤可能通过抑制bcl-2的表达,同时促进bax的表达,调节bax/bcl-2比值来发挥对RA-FLS的促凋亡作用,具有治疗RA的潜在价值。Objective To observe the effects of alcohol extract of Kunmu decoction（KMT）on the expression of B-cell lymphoma-2（bcl-2） and bcl-2 associated X protein（bax） in human rheumatoid arthritis fibroblast-like synoviocytes（RA-FLS）in vitro,and to explore the possible therapeutic mechanism. Methods Synovial tissues were obtained from patients with active RA subjecting to the revised criteria of the American College of Rheumatology,who received joint replacement or arthroscopy. Real time-PCR was used for the expression of bax and bcl-2 m RNA in RA-FLS.Enzyme-linked immunosorbent assay（ELISA） was used to detect the expression of bax and bcl-2 protein. Results High-dosage KMT had obvious inhibitive effect on the expression of bax induced by tumor necrosis factor alpha（TNF-α）（P〈0.01）. Compared with TNF-α group,methotrexate（MTX） and high-dosage KMT had obvious inhibitory effect on the expression of bcl-2（P〈0.05,P〈0.01）. Compared with MTX group,high-dosage KMT had obvious inhibitory effect on the expression of bcl-2 m RNA and protein（P〈0.05, P〈0.01）. Low-, moderate- and high-dosage KMT increased the ratio of bax/bcl-2 in a dose-dependent manner. Conclusion K unmu decoction may promote the apoptosis of RA-FLS by regulating the expression of bcl-2, bax m RNA and protein and the ratio of bax/bcl-2,showing possible therapeutic effect for RA.

关 键 词：昆母汤 类风湿关节炎 成纤维样滑膜细胞 B淋巴细胞瘤-2基因 bcl-2相关X蛋白 

分 类 号：R285.5[医药卫生—中药学]