摘要
目的:本实验通过研究黄芪甲苷对肿瘤坏死因子α(tumor necrosis factor-α,TNF-α)诱导的人类风湿关节炎成纤维样滑膜细胞(human fibroblast-like synoviocytes,HFLS-RA)增殖作用的影响,探讨其调控HFLS-RA内白细胞介素6(Interleukin-6,IL-6)、c-Jun氨基末端激酶1(JNK1)、c-Jun氨基末端激酶1/2(JNK1/2)蛋白表达的作用机制。方法:体外培养HFLS-RA,采用TNF-α及不同浓度的黄芪甲苷(astragaloside IV,AS-Ⅳ)干预细胞。本次实验共分为空白组(10%胎牛血清培养基培养细胞)、TNF-α组(10 ng/mL TNF-α)、AS-ⅣH组(10 ng/mLTNF-α+100μg/mLAS-Ⅳ)、AS-ⅣM组(10 ng/mLTNF-α+50μg/mLAS-Ⅳ)、AS-ⅣL组(10 ng/mLTNF-α+25μg/mL AS-Ⅳ)5组,MTT法检测不同浓度的黄芪甲苷对TNF-α诱导的HFLS-RA增殖活力的影响,ELISA法检测细胞上清液中IL-6的表达水平,Western Blot法检测细胞中JNK1、JNK1/2蛋白表达水平。结果:黄芪甲苷能够抑制TNF-α诱导的HFLS-RA细胞增殖,降低HFLS-RA中IL-6、JNK1、JNK1/2蛋白表达。结论:黄芪甲苷通过调控JNK信号通路相关细胞因子表达抑制TNF-α诱导的HFLS-RA细胞增殖。
Objective:To study the proliferous inhibiting effect of Astragaloside IV on human rheumatoid arthritis fibroblast-like synoviocytes(HFLS-RA)reduced by tumour necrosis factor(TNF-α),and investigate the molecular mechanism of Astragaloside IV in regulating IL-6,JNK1,and JNK1/2 proteins in HFLS-RA cells.Methods:HFLS-RA was cultured in vitro,the cells were treated with TNF-αand different concentrations of astragaloside.The experiment was divided into 5 groups:Blank group:culture medium containing 10%fetal bovine serum;TNF-αgroup:10 ng/mL TNF-α;AS-ⅣH group:10 ng/mLTNF-α+100μg/mLAS-Ⅳ;AS-ⅣM groups:10 ng/mL TNF-α+50μg/mLAS-Ⅳ;AS-ⅣL groups:10 ng/mLTNF-α+25μg/mLAS-Ⅳ,the proliferation of HFLS-RA was detected by MTT assay.The expression of IL-6 in the supernatant was detected by ELISA.The JNK1 and JNK1/2 proteins were detected by Western Blot.Results:Astragaloside IV can inhibit the proliferation of HFLS-RA induced by TNF-αand decrease the expression of IL-6,JNK1 and JNK1/2 in HFLS-RA.Conclusion:Astragaloside IV inhibits the proliferation of HFLS-RA cells which is reduced by TNF-αby regulating the expression of cytokines related to the JNK signaling pathway.
作者
冯美杰
李蕾
王颖航
徐兢鸿
潘志
FENG Mei-jie;LI Lei;WANG Ying-hang;XU Jing-hong;PAN Zhi(Changchun University of Chinese Medicine,Changchun 130117,China;Affiliated Hospital of Changchun University of Chinese Medicine,Changchun 130021,China)
出处
《中国中医基础医学杂志》
CAS
CSCD
北大核心
2020年第10期1484-1487,共4页
JOURNAL OF BASIC CHINESE MEDICINE
基金
国家自然科学基金资助项目(81273651)-从ERK5信号通路探讨消癥活络方抑制肝星状细胞活化机制的研究
国家自然科学基金资助项目(81374025)-SOCS-1在糖尿病肾病发生中的作用及荔枝核活性成分对其干预机制的研究
吉林省中医药科技项目(2018041)-当归补血汤防治RA的临床前基础研究。
