摘要
目的:探讨左归丸防治骨质疏松症的作用机制。方法:用含或不含p38 MAPK特异性阻滞剂(SB203580)的孵育液预处理细胞60min,加入左归丸含药血清孵育48h后,采用MTT法检测细胞活力;采用蛋白质印迹分析方法测定p38活性和Runx2蛋白表达水平;采用实时定量PCR方法测定Runx2和Col I mRNA表达水平。结果:左归丸含药血清能够显著增加细胞活力和磷酸化p38蛋白表达水平,诱导Runx2和Col I表达(P<0.01)。SB203580抑制了左归丸含药血清诱导的细胞活力和p38蛋白磷酸化水平,同时下调了左归丸含药血清诱导的Runx2和Col I表达(P<0.01)。结论:左归丸含药血清可能部分通过活化p38 MAPK信号通路,上调Runx2表达,刺激Col表达,促进骨形成。
Objective： To explore the mechanism of Zuogui Pill （ZGP） to prevent and treat osteoporosis. Methods： MC3T3 cells were preincubated with or without a specific inhibitors （SB203580） of p38 mitogen activated protein kinase （MAPK） for 60min, and the cell viability was assayed by MTT method after the cell cultured with ZGP pharmacological serum for 48h. p38 MAPK activity and Runx2 protein expression were assayed by Western Blot analysis. Runx2 and type I collagen （Col I） mRNA expression were evaluated by the method of real-time quantitative reverse transcriptase polymerase chain reaction （qRT-PCR）. Results： Cell viability and phosphorylated level of p38 could be increased by ZGP pharmacological serum significantly （P〈0.01）. ZGP pharmacological serum induced expression of Runx2 and Col I （P〈0.01）. ZGP-induced cell viability and phosphorylated level of p38 were repressed by SB203580. ZGP-induced expression of Runx2 and Col I were also decreased by SB203580 （P〈0.01）. Conclusion： The osteogenic functions of ZGP pharmacological serum might partly develop through activation of p38 signaling pathway to up-regulate the expression of Runx2, and stimulate Col I expression.
出处
《中华中医药杂志》
CAS
CSCD
北大核心
2013年第5期1457-1461,共5页
China Journal of Traditional Chinese Medicine and Pharmacy
基金
National Natural Science Foundation of China(No.30873226)
Innovation Team Project of Liaoning Provincial Department of Education(No.LT2010068)
Support Programs of Colleges and Universities in Liaoning Province Talents(No.LR201025)
Science and Technology Department of Liaoning Province Natural Science Foundation Project Plan(No.201102148)~~
