逍遥丸对小鼠卵母细胞BMP-6/Smads通路表达的影响

Effect of Xiaoyao Pill on the Expression of BMP-6/Smads Signal Pathway in Mouse Oocytes

目的:观察逍遥丸对小鼠卵母细胞BMP-6、BMPR II、ALK-2、ALK-6、Smad1、Smad5、Smad8和Smad4表达的影响。方法:将200只雌性小鼠按随机数字表法分为疏肝高、低剂量组、COH组和空白对照组4组各50只。应用免疫组化和real-time PCR方法分别测定卵母细胞中BMP-6、BMPR II、ALK-2、ALK-6、Smad1、Smad5、Smad8和Smad4蛋白和mRNA的表达。结果:与空白对照组比较,COH组BMP-6、BMPR II、ALK-2、ALK-6、Smad1、Smad5、Smad8和Smad4蛋白和mRNA表达差异无统计学意义,各治疗组BMP-6、ALK-2、ALK-6、Smad1、Smad5、Smad8和Smad4蛋白和mRNA表达增加,疏肝高剂量组优于低剂量组;疏肝高剂量组BMPR II蛋白和mRNA表达增加。结论:逍遥丸可使BMP-6表达上调,激活其Ⅱ型受体BMPRⅡ形成受体复合物,然后募集Ⅰ型受体ALK-2和ALK-6使其活化,活化的Ⅰ型受体进一步磷酸化细胞内信号Smad1/5/8,然后与Smad4结合,从而启动信号传导干预卵泡发育和卵母细胞的质量。

Objective： To investigate the effects of Xiaoyao Pill on the expression of BMP-6,BMPR II,ALK-2,ALK-6,Smad1,Smad5,Smad8 and Smad4 in mouse oocytes. Methods： 200 mice were randomly divided into 4 groups： Shugan high dose group,Shugan low dose group,COH group and control group（ N = 50）,the expression of BMP-6,BMPR II,ALK-2,ALK-6,Smad1,Smad5,Smad8 and Smad4 in mouse oocytes were detected by immunohistochemistry and Real-time PCR.Results： Compared with the control group,the protein content and mRNA expression of BMP-6,BMPRII,ALK-2,ALK-6,Smad1,Smad5,Smad8 and Smad4 in the COH group have no statistical significance. the protein content and mRNA expression of BMP-6,ALK-2,ALK-6,Smad1,Smad5,Smad8 and Smad4 in the treatment groups increased. The protein and mRNA expression in Shugan high dose group were higher than the Shugan low group. The protein and mRNA expression of BMPR II in Shugan high dose group were increased. Conclusion： Xiaoyao Pill can improve the expression of BMP-6 in mouse oocytes,activate its second type of receptor BMPRⅡ,and then activate receptors ALK-2 /6,the acitvation of type 1receptors phosporylate intracellular signal Smad1 /5 /8,then combine with Smad4,and start the signal transduction of Smads pathway to intervene the development and quality of oocytes..