通精灵对精索静脉曲张模型大鼠生精细胞凋亡及线粒体超微结构的影响

Effect of Tongjingling Prescription on Apoptosis and Mitochondrial Ultrastructure of Spermatogenic Cells in Testis of Rats with Experimental Varicocele

目的:探讨通精灵对实验性精索静脉曲张大鼠生精细胞凋亡与线粒体超微结构的影响以及其部分作用机制。方法:雄性Wistar大鼠75只随机分为假手术组、模型组、高剂量组、中剂量组、低剂量组,每组15只。制备大鼠精索静脉曲张模型,造模完成后4周开始给药,持续8周。观察大鼠的一般情况;Annexin V-FITC方法检测各组生精细胞凋亡;免疫组化检测Cyt-c蛋白表达;透射电镜观测各组生精细胞线粒体超微结构。结果:与假手术组比较,模型组生精细胞凋亡率明显升高(P<0.01)。与模型组相比较,通精灵各组凋亡率均有下降,差异具有显著性(P<0.01)。Cyt-c在间质细胞和各级生精细胞的细胞质中可见棕黄色表达,以精母细胞较明显。与假手术组相比较,模型组Cyt-c表达升高(P<0.01)。与模型组比较,通精灵各组Cyt-c表达降低(P<0.01)。透射电镜结果显示模型组生精细胞线粒体数目减少,超微结构发生改变,线粒体损伤。通精灵各组线粒体存在不同程度的超微结构的改变,但改变程度与模型组相比较轻。结论:精索静脉曲张模型大鼠生精细胞凋亡率明显升高,线粒体超微结构改变,线粒体损伤明显,生精细胞Cyt-c蛋白表达升高,通精灵可能是通过保护生精细胞线粒体超微结构,抑制生精细胞Cyt-c蛋白表达,从而调节线粒体途径引起的精索静脉曲张大鼠生精细胞过度凋亡。

Objective:To explore the effects of Tongjingling on apoptosis and mitochondrial ultrastructure of spermatogenic cells in the testis of rats with experimental varicocele.Methods:Seventy-five Wistar male rats were divided into five groups:sham operation group,model group,Tongjingling Prescription high dose group,middle dose group and low dose group,15 mice of each group.Experimental varicocele was created in the rats by partial ligation of left renal vein.Four weeks after establishing the model,medicine was given for 8 weeks.We observed the general situation of rats,apoptosis was examined by flow cytometric analysis and the expression of Cyt-c was measured by Immunohistochemical analysis.The mitochondrial ultrastructure of spermatogenic cells was examined with transmission electron microscope.Results:Compared with sham operation group,the apoptosis rate was significantly increased in model group(P<0.01).Compared with model group,the apoptosis rate was significantly reduced in Tongjingling groups(P<0.01).Cyt-c content in model group was significantly higher than that of the sham operation group(P<0.05),also the cyt-c content in Tongjingling groups but still significantly lower than that of the model group(P<0.05).Microscopic examination showed less damage in Tongjingling group than that in model group.Conclusion:The apoptosis rate was significantly increased,microscopic examination damage and Cyt-c content increased in the testis of rats with experimental varicocele.There is significantly increased apoptosis,microscopic changes in mitochondria,damage of mitochondria and elevated Cyt-c protein expression in the testis of rats with experimental varicocele.So we infer that the apoptosis of spermatogenic cells is associated with damage of mitochondria.Tongjingling could reduce apoptosis of spermatogenic cells by reducing mitochondrial damage.