菟丝子提取物对肾阳虚大鼠睾丸P450arom、CYP19表达及性激素和精子的影响

Expression of P450arom CYP19 Gonadal Hormone Levels and Influence of the Number of Spermium in the Testis of Infertile Rats with Kidney-yang Deficiency after Treated by Extractive of Cuscuta

目的:观察菟丝子提取物作用于腺嘌呤诱导的肾阳虚不育大鼠睾丸中P450arom、CYP19的表达、激素含量及精子数量的变化,探讨肾虚引起的雄性不育的病理机制及菟丝子提取物的作用。方法:30只雄性大鼠随机分为3组:正常对照组、模型组、治疗组,每组10只。用腺嘌呤150mg/(kg.d),分别给模型组及治疗组大鼠灌胃30天,诱导肾阳虚不育模型。诱导模型成功后,模型组用生理盐水灌胃30天;用菟丝子提取物200mg/(kg.d)给治疗组灌胃30天;用免疫组化法和Western-blot检测P450arom及CYP19在大鼠睾丸中的表达;用扫描电镜及透射电镜观察曲细精管中精子数量的变化;精子自动检测仪扫描观察精子的密度;放射免疫的方法检测FSH、LH、T、E2含量。结果:CYP19在大鼠睾丸的间质细胞、长形精子细胞中均有表达,其免疫阳性物质位于间质细胞质和长形精子细胞的头、尾部;P450arom表达于睾丸间质细胞的细胞质中。模型组大鼠睾丸CYP19、P450arom则明显低于正常对照组(P<0.05),治疗组CYP19、P450arom的表达明显高于模型组(P<0.05),正常对照组与治疗组之间无显著差异(P>0.05);正常对照组及治疗组曲细精管中的精子数量及FSH、LH、T和E2的含量明显高于模型组,正常对照组与治疗组之间无显著差异(P>0.05)。结论:P450arom及CYP19在睾丸表达的异常可能与肾虚引起的雄性大鼠不育有关,菟丝子提取物能够增加肾虚大鼠睾丸中P450arom、CYP19的表达,提高精子的数量及FSH、T、E2含量,为进一步探讨P450arom及CYP19在雄性肾虚不育模型大鼠精子生成和成熟中的作用及菟丝子提取物的作用机制提供了实验依据。

Objective：To observe the expression of P450arom, CYP19 hormone levels and the number of spermium in the testis of infertile rats with adenine-modeled kidney-yang deficiency after treated by extractive of Cuscuta;to study the pathological mechanism of infertility with kidney-yang deficiency and the effect of Cuscuta Flavonoid. Methods ： Twenty 3 month male SD rats were divided randomly into 3 groups with 10 in each normal control, mode group, treatment group. The experimental rats had been fed with adenine （ 150mg · kg^-1 · d^-1 ）for 30 days except normal control. After been fed with adenine, treatment group was fed with extractive of Cuscuta for 30 days. The expression levels of P450 aromatase, CYP19 were measured with immunohistochemical SABC and Western-blot method, while the change of the number of spermium was observed by scanning electron microscope and transmission electron microscope technique. Use radio- immunity method to detect the levels of FSH, LH, T and E2. Results ： CYP19 was expressed in cell of Leydig, dolicho-nematoblast in rats＇ testis with their positive immuno-responsive substance locating in the cytoplasm but showed negative response in spermatogenic cell and Sertoli＇s cell; P450aromatase was positive expressed in cytoplasm of Leydig＇s cell but showed negative response in spermatogenic cell and Sertoli＇s cell. Compared with the normal control, P450aromatase and CYP19 expression were lower in mode group （P〈0.05）; P450aromatase and CYP19 expression were higher in the treatment group compared with mode group（P〈0.05 ）. Scanning electron microscope and transmission electron microscope showed that in treatment group the number of spermium increased significantly compared with mode group and radio-immunity method showed that FSH, LH, T and E2 in treatment group were higher than in mode group （P〈0.05）, hut there was no obvious difference between normal control and treatment group （P〉0.05）. Conclusion ： The abnormality expression P450arom, CYP19 in testis was possible with infertility with kidney-yang deficiency ; Cuscuta Flavonoid can improve the expression P450arom, CYP19, the number of spermium and the hormone levels. For further approach P450arom, CYP19 in infertility rats with kidney-yang deficiency gonepoiesis and correlation and the mechanism of the extractive of Cuscuta provided preliminary experimental evidence.