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2020, v.45(19) 4712-4718

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紫草素对类风湿关节炎成纤维样滑膜细胞功能的影响
Effect of shikonin on function of rheumatoid arthritis fibroblast like synoviocytes

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;Shenzhen Hospital, Peking University;

摘要(Abstract):

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-α could 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-α group, 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.

关键词(KeyWords): 紫草素;MH7A细胞;增殖;迁移;侵袭;黏附
shikonin;MH7A cells;proliferation;migration;invasion;adhesion

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金面上项目(81573674)

作者(Author): 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;Shenzhen Hospital, Peking University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200506.401

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