摘要
目的:研究黄芪甲苷对骨髓间充质干细胞(BMSCs)炎症模型的影响及分子机制。方法:采用全骨髓贴壁培养分离法获取小鼠BMSCs,并利用流式细胞术对特异分子标志(CD90和CD45)进行鉴定。脂多糖(LPS,500 ng·ml-1)处理BMSCs构建细胞炎症模型后,随机分为4组:单纯炎症模型组、黄芪甲苷低、中、高剂量组(100,500,1 000 ng·ml-1)。采用流式细胞术及ELISA检测细胞周期、凋亡比例及培养上清中细胞因子分泌情况;采用丝裂原活化蛋白激酶/细胞外调节蛋白激酶(MAPK/ERK)通路抑制剂U0126探究黄芪甲苷影响BMSCs的分子机制。结果:经3次传代的细胞属于CD90+/CD45-的BMSCs。与单纯炎症模型组相比,不同剂量黄芪甲苷处理后G1期细胞比例减少,细胞凋亡比例降低(P<0.05)。单纯炎症模型组的BMSCs可分泌白介素8(IL-8)、白介素1β(IL-1β)及肿瘤坏死因子-α(TNF-α),经黄芪甲苷处理后各细胞因子下降(P<0.05);加入MAPK/ERK通路抑制剂U0126后,IL-8、IL-1β及TNF-α水平高于黄芪甲苷处理组(500 ng·ml-1)(P<0.05),但与单纯炎症模型组相比差异无统计学意义(P>0.05)。结论:黄芪甲苷可能通过MAPK/ERK通路促进LPS诱导的BMSCs增殖,抑制其凋亡和促炎细胞因子的分泌。
Objective:To study the effect of asragalosideⅣon the inflammation model of bone mesenchymal stem cells(BMSCs)and the underlying mechanism.Methods:BMSCs were amplified and purified by the whole marrow adherent method,and flow cytometry(FCM)was used to detect the biomarker of mesenchymal stem cells(CD90 and CD45).The inflammation model of BMSCs was constructed by lipopolysaccharide(LPS,500 ng·ml-1),and then randomly divided into four groups:the simple inflammation model group,astragalosideⅣlow,medium and high dose groups(100,500 and 1000 ng·ml-1).FCM and ELISA were used to analyze the situation of cell cycle,apoptosis and cytokine secretion.The mitogen-activated protein kinase/extracellular regulated protein kinases(MAPK/ERK)inhibitors(U0126)were utilized to study the molecular mechanism of asragalosideⅣon BMSCs.Results:After three passages,the cells belonged to BMSCs of CD90+/CD45-type.Compared with those in the inflammation model group,the percentage of G1 cells and the apoptotic cells decreased after the treatment with different doses of astragalosideⅣ(P<0.05).The BMSCs in the simple inflammation model group secreted interleukin-8(IL-8),interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α),and the cytokines decreased after the treatment with astragalosideⅣ(P<0.05).After the MAPK/ERK inhibitors(U0126)were added,the levels of IL-8,IL-1βand TNF-αwere higher than those in astragalosideⅣgroup(500 ng·ml-1)(P<0.05),while there were no significant differences with those in the inflammation model group(P>0.05).Conclusion:AsragalosideⅣcan promote the proliferation,inhibit the apoptosis and promote the inflammatory cytokines excretion of LPS induced BMSCs through MAPK/ERK pathway.
作者
闾红燕
Lv Hongyan(Department of Pharmacy,Xiasha Branch,Sir Run Shaw Hospital,School of Medicine,Zhejiang University,Hangzhou 310000,China)
出处
《中国药师》
CAS
2019年第9期1625-1629,共5页
China Pharmacist
