基于HSP60、HSP90表达探讨续断种子方对少弱精子症模型大鼠精液质量的影响

Based on the expression of HSP60 and HSP90,to explore the effect of Xuduan Zhongzi prescription on semen quality of oligoasthenospermia model rats

目的:探讨续断种子方对少弱精子症模型大鼠睾丸生精细胞HSP60、HSP90表达及精液质量的影响。方法:将40只SPF级雄性大鼠随机分为空白对照组10只和模型组30只,造模成功后按照随机计数表法将模型组分为模型对照组、左卡尼汀组和续断按种子方组各10只。左卡尼汀组和续断种子方组大鼠根据人体等效计量给予对应药物灌胃,空白对照组和模型对照组给予等量生理盐水灌胃,连续给药8周后处死各组大鼠。取各组左侧附睾进行精子质量检测,取各组左侧睾丸采用HE染色方法观察组织形态学变化,采用免疫组化法和Western Blot法检测睾丸组织中HSP60和HSP90的表达情况。结果:HE染色结果可见,与空白对照组相比,模型对照组大鼠曲细小管明显萎缩退化且排列不规则,生精上皮呈大量空泡状,支持细胞变性,间质细胞内炎性浸润明显,间质细胞增生,上皮细胞散在排列,大量脱落变性,精子少见。续断种子方干预后以上病变得到明显改善,生精小管结构相对规整,排列有序,较模型对照组致密,支持细胞与间质细胞明显增多,生精上皮较模型组增厚,层次丰富,管腔内可见大量精子。免疫组化和Western Blot检测显示,模型大鼠经续断种子方干预后HSP60、HSP90表达量显著增加(P<0.01)。精子质量检测结果显示,与空白对照组相比,模型对照组精子浓度和活动率明显降低(P<0.01),经续断种子方干预后,模型大鼠精子浓度及活动率均有明显提高(P<0.01)。结论:续断种子方可能通过提高HSP60、HSP90表达量,抑制少弱精子症模型大鼠睾丸生精细胞凋亡,从而改善精子质量。

Objective:To investigate the effect of Xuduan Zhongzi prescription on the expression of HSP60 and HSP90 in testicular spermatogenic cells and semen quality of oligozoospermia and asthenospermia model rats.Methods:A total of 40 SPF male rats were randomly divided into 10 blank control group and 30 model group.After successful modeling,the model group was divided into model control group,levocarnitine group and Xuduan Zhongzi prescription group with 10 rats in each group.The rats in levocarnitine group and Xuduan Zhongzi prescription group were given corresponding drugs by gavage according to the equiva⁃lent measurement of humanbody,the blank control group and model control group were given the same amount of normal saline by gavage,and the rats in each group were killed after 8 weeks of continuous administration.Take the left epididymis of each group for sperm quality detection,take the left testis of each group,observe the histomorphological changes by HE staining,and detect the expression of HSP60 and HSP90 in testicular tissue by immunohistochemistry and Western blot.Results:HE staining showed that compared with the blank control group,the convoluted tubules in the model group were significantly atrophic,degenerated and irregularly arranged,the spermatogenic epithelium showed a large number of vacuoles,Sertoli cell degeneration,obvious in⁃flammatory infiltration in stromal cells,stromal cell proliferation,scattered rows of epithelial cells,a large number of exfoliation and degeneration,and sperm were rare.After the intervention of Xuduan Zhongzi prescription,the above lesions were significantly improved,the structure of seminiferous tubules was relatively regular and orderly,denser than that of the model group,sertoli cells and stromal cells increased significantly,the seminiferous epithelium was thicker than that of the model group,with rich lay⁃ers,and a large number of sperm could be seen in the lumen.Immunohistochemistry and Western Blot showed that the expression of HSP60 and HSP90 increased significantly after the intervention of Xuduan Zhongzi prescription in model rats(P<0.01).The sperm quality test results showed that compared with the blank control group,the sperm concentration and activity rate in the mod⁃el group decreased significantly,(P<0.01).After continuous seed cutting,the sperm concentration and activity rate of model rats were significantly increased,(P<0.01).Conclusion:Xuduan Zhongzi prescription may inhibit the apoptosis of testicular spermato⁃genic cells in oligoasthenospermia model rats by increasing the expression of HSP60 and HSP90,so as to improve sperm quality.