Aim To study the effects of Zuogui pill medicated serum via ERKl/2-dependent pathway on MC3T3-E1 cell gene expression. Methods Conjugated Estrogens Tablets(BML) as a positive control, the SD female rats were fed with high, medium or lowdose of Zuogui Pill suspension. Zuogui pill medicated serum was separated by abdominal aortic blood after 7 days' feeding. MTT assay was applied to test the effects of Zuogui pill medicated serum or PD98059 stimulation on the MC3T3-E1 cellular viability. The level of ERK1/2 protein phosphorylation and TGF-β1, Smad4 protein was analyzed with Western blot. The expression of Cbfαl ,COL I mRNA was detected by Real Time RT-PCR. Results Zuogui pill medicated serum and BML promoted MC3T3-E1 cell proliferation, inducedERK1/2 protein phosphorylation, increased TGF -β1, Smad4 protein production and Cbfal, COL I mRNA expression. The proliferation of MC3T3-E1 cell was decreased, ERK1/2 protein phosphorylation and Smad4 protein production were reduced, and gene expression except for TGF-β1 were down-regulated after the addition of PD98059. Conclusion Zuogui pill is effective in promoting osteoblast proliferation and differentiation. Zuogui pill may play an estrogen-like role through the regulation of the ERK/Smads signaling pathway, so as to carry the point of prevention and treatment of osteoporosis.%目的 研究ERK1/2信号通路在左归丸含药血清调控MC3T3-E1细胞基因表达中的作用.方法以倍美力为阳性对照药,对SD ♀大鼠灌服高、中、低剂量的左归丸混悬液,d 7腹主动脉取血分离含药血清.采用MTT法检测左归丸含药血清和PD98059对MC3T3-E1细胞增殖作用的影响,采用Western blot法检测ERK1/2蛋白的磷酸化水平及TGF-β1、Smad4蛋白表达,采用Real Time RT-PCR法检测Cbfα1、COLⅠmRNA表达.结果 左归丸含药血清和倍美力能促进MC3T3-E1细胞增殖,诱导ERK1/2蛋白的磷酸化,促进TGF-β1、Smad4蛋白分泌,上调其Cbfα1、COLⅠmRNA表达;加入PD98059后MC3T3-E1细胞增殖下降、ERK1/2蛋白的磷酸化水平降低、TGF-β1 蛋白表达进一步升高、Smad4蛋白表达降低、Cbfα1、COLⅠmRNA表达下调.结论 左归丸能有效促进成骨细胞增殖和分化;左归丸可能是通过发挥雌激素样作用调控了ERK/Smads信号通路,从而达到防治骨质疏松的目的.
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