Lian-Hua H E, Qian-Qian Wang, Cong-Cong Sun, Na Lin, Chun-Fang Liu
Institute of Chinese Materia Medica, China Academy of Chinese Medical Scineces Beijing 100700, China Shenzhen Hospital, Peking University Shenzhen 518036, China.
Institute of Chinese Materia Medica, China Academy of Chinese Medical Scineces Beijing 100700, China.
Zhongguo Zhong Yao Za Zhi. 2020 Oct;45(19):4712-4718. doi: 10.19540/j.cnki.cjcmm.20200506.401.
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.
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）成纤维样滑膜细胞增殖、迁移、黏附和侵袭的影响，并从蛋白激酶B（Akt）和丝裂原活化蛋白激酶（MAPK）信号通路方面探讨其作用机制。本实验用TNF-α（20 ng·mL⁻¹）诱导人RA成纤维样滑膜细胞系（MH7A）。加入不同浓度的紫草素（0.025、0.05、0.1 pmol·L⁻¹）后，分别采用MTT法、划痕试验、黏附试验、Transwell侵袭试验检测MH7A细胞的增殖、迁移、黏附和侵袭能力。采用蛋白质免疫印迹法检测MH7A细胞中Akt和MAPK信号通路分子的蛋白表达。结果显示，与对照组相比，TNF-α可显著诱导MH7A细胞的增殖、迁移、黏附和侵袭，并增加Akt、JNK、p38和细胞外调节蛋白激酶（ERK）的磷酸化水平。与TNF-α组相比，紫草素处理24 h后对TNF-α诱导的MH7A细胞增殖无显著影响，处理48 h后可呈浓度依赖性降低TNF-α诱导的MH7A细胞增殖。紫草素还可在24 h内显著降低TNF-α诱导的MH7A细胞的迁移、黏附、侵袭以及Akt、JNK、p38、ERK的磷酸化水平。这些结果表明，紫草素可降低TNF-α诱导的MH7A细胞的增殖、迁移、黏附和侵袭能力，其机制可能与下调Akt和MAPK信号通路的激活有关。
