摘要
为了观察紫草素对肿瘤坏死因子-α(TNF-α)诱导的类风湿关节炎(RA)成纤维样滑膜细胞的增殖、迁移、黏附和侵袭能力影响,并从蛋白激酶B(Akt)和丝裂原活化蛋白激酶(MAPK)信号通路探索其作用机制,该实验采用TNF-α(20 ng·mL-1)体外诱导RA患者成纤维样滑膜细胞系(MH7A),加入不同浓度紫草素(0.025,0.05,0.1 pmol·L^-1)作用后,分别采用四甲基偶氮唑蓝比色法、划痕试验、黏附试验及Transwell侵袭试验检测MH7A细胞的增殖活性以及迁移、黏附和侵袭能力,蛋白免疫印迹法检测MH7A细胞中Akt和MAPK信号通路分子的蛋白表达水平。结果显示,与对照组相比,TNF-α能明显诱导MH7A细胞的增殖、迁移、黏附和侵袭能力,升高Akt,c-Jun氨基末端激酶(JNK),p38及细胞外调节蛋白激酶(ERK)的磷酸化水平;与TNF-α组相比,紫草素作用24 h对TNF-α诱导的MH7A细胞增殖活性没有明显影响,作用48 h能浓度依赖地降低TNF-α诱导增加的MH7A细胞增殖活性,作用24 h内还能显著降低TNF-α诱导异常升高的MH7A细胞迁移、黏附、侵袭能力及Akt,JNK,p38,ERK的磷酸化水平。这些结果提示紫草素可降低TNF-α诱导异常升高的MH7A细胞增殖、迁移、黏附和侵袭能力,其机制可能与下调Akt和MAPK信号通路的活化相关。
To observe the effect of shikonin on the proliferation,migration,adhesion and invasion of rheumatoid arthritis(RA)fibroblast like synoviocytes induced by tumor necrosis factor-α(TNF-α),and to explore its mechanism of action from aspects of protein kinase B(Akt)and mitogen activated protein kinase(MAPK)signaling pathways.TNF-α(20 ng·mL^-1)was used in this experiment to induce human RA fibroblast like synovial cell line(MH7 A).After addition of different concentrations of shikonin(0.025,0.05,0.1 pmol·L^-1),the proliferation,migration,adhesion and invasion ability of MH7 A cells were detected by MTT test,scratch test,adhesion test,Transwell invasion test,respectively.Protein expression of Akt and MAPK signaling pathway molecules in MH7 A cells was detected by Western blot.The results showed that as compared with the control group,TNF-αcould significantly induce the proliferation,migration,adhesion and invasion of MH7 A cells,and increase the phosphorylation level of Akt,JNK,p38 and extracellular regulatory protein kinase(ERK).As compared with the TNF-αgroup,shikonin had no significant effect on TNF-α-induced proliferation of MH7 A cells after 24 h treatment,and it could reduce the TNF-α-induced proliferation of MH7 A cells in a concentration dependent manner after 48 h treatment.Shikonin also significantly reduced the TNF-α-induced migration,adhesion,invasion and phosphorylation levels of Akt,JNK,p38,ERK in MH7 A cells within 24 h.These results suggested that shikonin could reduce the proliferation,migration,adhesion and invasion ability of MH7 A cells induced by TNF-α,and its mechanism may be related to the down-regulation of Akt and MAPK signaling pathway activation.
作者
何莲花
汪倩倩
孙丛丛
林娜
刘春芳
HE Lian-hua;WANG Qian-qian;SUN Cong-cong;LIN Na;LIU Chun-fang(Institute of Chinese Materia Medica,China Academy of Chinese Medical Scineces,Beijing 100700,China;Shenzhen Hospital,Peking University,Shenzhen 518036,China)
出处
《中国中药杂志》
CAS
CSCD
北大核心
2020年第19期4712-4718,共7页
China Journal of Chinese Materia Medica
基金
国家自然科学基金面上项目(81573674)。
关键词
紫草素
MH7A细胞
增殖
迁移
侵袭
黏附
shikonin
MH7A cells
proliferation
migration
invasion
adhesion
作者简介
何莲花,助理研究员,主要从事中药抗类风湿关节炎相关研究,E-mail:helianhua126@126.com;通信作者:刘春芳,副研究员,主要从事中药抗炎免疫药理研究,E-mail:cfliu@icmm.ac.cn。
