温阳生精汤对肾阳虚不育大鼠睾丸Smads基因表达的调控

Regulatory Effect of Wenyang Shengjing Decoction on Smads Expressions in Testis of Sterile Rats with Shen-yang Deficiency

目的探讨转化生长因子-β(transforming growth factor-β,TGF-β)超家族的细胞内信号内转导分子Smads(mother against dpp)基因腺嘌呤致雄性肾阳虚不育大鼠睾丸中表达的定位和分布及温阳生精汤对Smads基因表达的调控作用。方法将实验大鼠随机分为3组,分别为正常对照组、肾阳虚不育组和温阳生精汤组。应用免疫组织化学SABC法和半定量RT-PCR法检测实验大鼠睾丸中Smad1、Smad2、Smad4的定位和分布,并通过图像分析系统进行统计学分析;应用放射免疫法测定血清中睾酮(testosteone,T)、促黄体激素(luteinizing hormone,LH)和促卵泡激素(follicle-sti mulating hormone,FSH)含量;同时测定大鼠体重、睾丸和附睾重量及精子数量。结果Smad1和Smad2在实验大鼠睾丸的各级生精细胞中均有表达,其免疫反应阳性物质位于细胞质内,Smad2在支持细胞的胞质内亦可见阳性表达,两者在间质细胞呈阴性反应;Smad4在实验大鼠睾丸间质细胞的胞质内有表达,生精细胞及支持细胞呈阴性反应。肾阳虚不育组大鼠睾丸中Smad1的表达明显低于正常对照组(P<0.05),Smad2和Smad4的表达明显强于正常对照组(P<0.05)。温阳生精汤能明显增加Smad1在肾阳虚不育大鼠睾丸中的表达(P<0.05),降低Smad2和Smad4的表达(P<0.05)。与正常对照组比较,肾阳虚不育组大鼠体重明显下降,附睾的精子数量明显减少;血清中T显著降低,FSH、LH增高(P<0.05);而各实验组大鼠睾丸及附睾重量无明显变化(P>0.05);温阳生精汤组大鼠体重和精子数量较肾阳虚不育组显著增加;血清T水平增加,FSH、LH水平降低(P<0.05)。结论温阳生精汤不仅能够通过提高肾阳虚不育大鼠血清中T水平,降低FSH和LH水平,增加大鼠的精子数量;而且可以通过调控大鼠睾丸中TGF-β超家族的细胞内信号转导分子Smads基因的表达水平,直接或间接地影响T、FSH和LH水平,促进精子生成,达到治疗男性不育的目的。

Objective To study the localization and distribution of expressions of Smads （mother against dpp）, the intracellular signal transduction molecules in the transforming growth factor-β （TGF-β） family, in the testis of male sterile rats with Shen-yang deficiency induced by adenine and to observe the effect of Wenyang Shengjing Decoction （WSD） on these expressions. Methods Rats were randomly divided into the control group, the model group and the WSD group. Localization and distribution of Smad 1, Smad 2 and Smad 4 expressions were detected by immunohistochemistry SABC and semi-quantitative RT-PCR and analyzed statistically by image analysis system; the contents of testosterone （T）, luteinizing hormone （LH）, and foUicle-stimulating hormone （FSH） were detected by radioimmunoassay; and the weights of body, testis and epididymis, as well as sperm number of rats were also measured. Results Smad 1 and Smad 2 were expressed in cytoplasm of all levels of spermatogenic cells in rats＇ testis with their positive immuno-responsive substance locating in the cytoplasm, and positive Smad 2 expression could also be found in cytoplasm of Sertoli＇s cell, but both of them Smad 4 was positively expressed in cytoplasm of Leydig＇s cell but showed negative response in spermatogenic cell and Sertoli＇s cell. Compared with the normal control, Smad 1 expression was lower （P 〈 0.05）, but Smad 2 and Smad 4 were higher in the model group （both P 〈 0.05）, these abnormal changes could be reversed by WSD treatment （all P〈0.05）. Compared with the normal control group, the body weight, sperm number and serum T level were lower, and levels of FSH and LH were higher （all P〈0.05） in the model group, which could all be improved by WSD （P〈0.05） ; the weights of testis and epididymis were unchanged in all groups （P 〉 0.05 ）. Conclusion WSD could not only increase the sperm number through elevating serum T level and decreasing the levels of FSH and LH, but also by way of regulating Smads genes expression to adjust the levels of sex hormones, promote the production of sperm directly or indirectly, so as to treat male infertility.