五子衍宗丸对幼年大鼠睾丸支持细胞增殖的影响及机制

Influence and mechanism of Wuzi Yanzong Wan on proliferation of Sertoli cells in infant rats

目的探讨五子衍宗丸对幼年大鼠睾丸支持细胞的增殖作用及其机制。方法分离并培养20 d龄SD雄性大鼠的睾丸支持细胞,将支持细胞随机分为正常组和五子衍宗丸低、中、高剂量(5、10、15 g/L)组。采用MTT比色法检测各组支持细胞OD值的变化,采用免疫组化的方法检测各组支持细胞中Ki67阳性颗粒的表达,采用Western Blot方法检测各组支持细胞中Akt和p-Akt的表达。结果与正常组相比,经五子衍宗丸处理后的睾丸支持细胞增殖能力明显增加(P<0.05或P<0.01);Ki67阳性颗粒的表达明显增多(P<0.05或P<0.01);p-Akt的相对表达量明显增加(P<0.05或P<0.01)。结论五子衍宗丸可能通过激活Akt信号通路,上调p-Akt的表达,促进幼年睾丸支持细胞的增殖。

ObjectiveTo discuss the effect and mechanism of Wuzi Yanzong Wan （Five Seeds for Multiplying Pills） in the proliferation of Sertoli cells in infant rats. Methods The Sertoli cells were separated from male SD rats （20 d old） and cultured, and then randomly divided into normal group, lowdose Wuzi Yanzong Wan group （5 g/L, low-dose group）, mid-dose Wuzi Yanzong Wan group （10 g/L, mid-dose group） and high-dose Wuzi Yanzong Wan group （15 g/L, high-dose group）. The changes of optical density （OD） of Sertoli cells were detected by using methyl thiazolyl tetrazolium test （MTT） in all groups. The expression of Ki67 positive granules in Sertoli cells were detected by using immunohistochemistry technique in all groups. The expressions of Akt and p-Akt in Sertoli cells were detected by using Western blot method. Results Compared with normal group, the proliferation of Sertoli cells treated with Wuzi Yanzong Wan was promoted significantly （ P 〈 0.05 or P 〈 0.01 ）. The expression of Ki67 positive granules increased significantly （P 〈 0.05 or P 〈 0. 01 ）, and relative expression of p-Akt increased significantly （ P 〈 0. 05 or P 〈 0.01 ）. Conclusion Wuzi Yanzong Wan can possibly promote the proliferation of Sertoli ceils in infant rats through activating Akt signal transduction pathway and up-regulating p-Akt expression.