中文摘要：

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,d7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。

英文摘要：

Aim To study the effects of Zuogui pill medicated serum via ERK1/2-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 Cbfα1,COLⅠ mRNA was detected by Real Time RT-PCR.Results Zuogui pill medicated serum and BML promoted MC3T3-E1 cell proliferation,induced ERK1/2 protein phosphorylation,increased TGF-β1,Smad4 protein production and Cbfα1,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.