Piwil2及Stat3、Bcl-2蛋白在雄性不育小鼠睾丸组织中表达?

Expression of Piwil2,Stat3 and Bcl-2 Proteins in Infertile Mouse Testis

本研究主要是探讨Piwil2、Stat3、Bcl-2蛋白在不育的雄性小鼠中的表达量及三者之间的表达位置相关性。取雄性昆明种小鼠60只,随机分为实验组与对照组,每组30只。实验组采用雷公藤多苷药物灌胃造小鼠不育模型28 d;对照组按照同样剂量的生理盐水进行灌胃,持续时间及频次同实验组。造模后将两组雄性小鼠分别与雌性小鼠交配;再处死雄性小鼠取出两组的睾丸组织,采用免疫组织化学染色法和蛋白印迹检测法分别检测样本中Piwil2、Stat3及Bcl-2蛋白的表达状况。将实验组与对照组的检测结果进行比较,观察两组的蛋白表达差异性及三个蛋白表达的相关性。H.E染色显示,实验组小鼠睾丸组织生精小管结构与对照组相比,明显被破坏,精原细胞及初次级精母细胞数量明显减少,结合与雌鼠交配后受精能力明显下降的结果,说明不育造模成功。免疫组化(IHC)染色结果显示,实验组Piwil2、Stat3及Bcl-2蛋白的染色程度及阳性细胞数均明显低于对照组(P <0.01)。Western Blot结果同样显示,三种蛋白在实验组的表达量明显低于对照组(Piwil2蛋白P <0.05,Stat3蛋白P <0.05,Bcl-2蛋白P <0.01)。本研究说明,Piwil2、Stat3及Bcl-2蛋白在雄性不育小鼠中表达量均显著降低,这三个蛋白对小鼠精子生成及小鼠不育的发生起到重要调节作用。

We aimed to investigate the expression of Piwil2,Stat3 and Bcl-2 proteins and their location correlation in sterile male mice.Methods:60 male Kunming mice were randomly divided into the experimental group and the control group,30 in each group.The sterile mice were modeled with administration of Chinese herb Tripterygium wilfordii for 28 d.The control group was intragastrically administered with the same dose of normal saline for the same duration.After modeling,two groups of male mice were mated with female mice respectively.The samples are respectively detected for expressions of Piwil2 protein,Stat3 protein and Bcl-2 protein by immunohistochemical staining and Western blot.We compared the expressions of these three proteins between the two groups.The HE staining showed that the structure of seminiferous tubules in the testis tissue of the experimental group was significantly damaged,the number of spermatogenetic cells including spermatogonia and primary spermatocytes was significantly reduced,and the fertilization ability was significantly decreased after mating,indicationg that the model establishment was successful.The results of IHC staining showed that the expressions of Piwil2 protein,Stat3 protein and Bcl-2 protein and the number of positive cells in the experimental group were significantly lower than those in the control group(P<0.001)(Table 2).Western blot results also showed significantly decreased expressions of these three proteins in the experimental group than in the control group(Piwil2 protein:P<0.05,Stat3 protein:P<0.05,Bcl-2 protein:P<0.01)(Fig.3,Fig.4).This study shows that the expressions of Piwil2,Stat3 and Bcl-2 proteins in the testis may play important roles in the occurrence of infertility.