摘要
目的探讨蒲公英甾醇对类风湿关节炎成纤维样滑膜细胞(MH7A)的增殖、迁移和凋亡及TLR4/NF-κB信号通路的影响和机制研究。方法CCK-8检测蒲公英甾醇的毒性;ELISA法检测MH7A细胞中炎性介质的分泌水平;EDU检测MH7A细胞的增殖能力;Transwell法检测MH7A细胞的迁移能力;细胞流式术检测MH7A细胞的凋亡能力;Western blot法检测MH7A细胞中TLR4、p-NF-κB p65和NF-κB p65蛋白的表达情况。结果蒲公英甾醇的最佳作用浓度为5、10、15μg/ml;经LPS诱导后,蒲公英甾醇对MH7A细胞炎性因子的分泌量有抑制作用(P<0.05);与LPS组比较,不同浓度的蒲公英甾醇使MH7A细胞的增殖能力和迁移能力下降,凋亡率提高(P均<0.05);Western blot法检测结果显示,与LPS组比较,不同浓度蒲公甾醇使TLR4蛋白表达降低,NF-κB p65的磷酸化程度降低(P均<0.05)。结论蒲公英甾醇能降低MH7A细胞的增殖能力、迁移能力并诱导其凋亡,可能与其降低TLR4和NF-κB p65的磷酸化程度有关。
Objective To investigate the effect and mechanism of taraxasterol on the proliferation,migration and apoptosis of fibroblast-like synoviocytes(MH7A)and the TLR4/NF-κB signaling pathway in rheumatoid arthritis.MethodsCCK-8 was ued to detect the toxicity of taraxasterol.ELISA was used to detect the secretion of inflammatory mediators in MH7A cells.EDU was used to detect the proliferation capacity of MH7A cells.Transwell was used to detect the migration capacity of MH7A cells.Cell flow cytometry was used to detect the apoptotic capacity of MH7A cells.Western blot was used to detect the expression of TLR4,p-NF-κB p65 and NF-κB p65 proteins in MH7A cells.ResultsThe optimal concentration of taraxasterol is 5,10,15μg/ml.After LPS induction,taraxasterol inhibited the secretion of inflammatory factors in MH7A cells(P<0.05).Compared with the LPS group,taraxasterol at different concentrations decreased the proliferation and migration capacity of MH7A cells and increased the apoptosis rate(all P<0.05).Western blot results showed that compared with the LPS group,taraxasterol at different concentrations reduced the expression of TLR4 protein and reduced the degree of phosphorylation of NF-κB p65(all P<0.05).ConclusionTaraxasterol can reduce the proliferation,migration and induce apoptosis of MH7A cells,which may be related to the reduction of the phosphorylation of TLR4 and NF-κB p65.
作者
万海山
代伟宏
黄泽晓
吴开弟
曹国良
何和与
Wan Haishan;Dai Weihong;Huang Zexiao(Department of Emergency Traumatology,Second Affiliated Hospital of Hainan Medical College,Hainan 570311,China)
出处
《医学研究杂志》
2020年第11期126-131,共6页
Journal of Medical Research
基金
海南省卫生计生行业科研项目(1801320712A2080)。
