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目的初步研究自噬在精索静脉曲张大鼠模型中的表达改变和可能的理论意义,及初步探讨自噬和凋亡的可能关系,为精索静脉曲张致不育机制的研究提供新的方向。方法取SD雄性大鼠40只,分为正常对照组、VC组(VC造模组)、VC手术组(VC+手术治疗组)、假手术组,每组10只。正常对照组不做任何处理,VC组和VC手术组行造模,假手术组打开腹腔但不做任何处理,第30天时对VC手术组行左侧精索静脉高位结扎手术治疗。第60天时对所有组大鼠行左侧睾丸切除术。对睾丸组织行HE染色、TUNEL染色,透射电镜观察用于睾丸生精小管上皮细胞自噬体;Western-blot、RT-PCR检测LC3-Ⅱ和Beclin1含量变化。结果透射电镜观察正常对照组和假手术组生精小管上皮细胞中较少见自噬体和自噬溶酶体,VC组和VC手术组中自噬体、自噬溶酶体明显增多。Western-blot及RT-PCR检测示,Beclin-1和LC3-Ⅱ/Ⅰ在VC组和VC手术组中表达明显高于正常对照组和假手术组。结论 VC可致大鼠睾丸生精小管上皮细胞中自噬体表达增多的现象,并且初步探讨自噬与凋亡呈正相关的关系。对VC睾丸细胞自噬的研究,可能进一步为VC的临床治疗提供新的靶点和理论依据。
Objective To study the change of autophagy in rat model of varicocele and its possible theoretical significance, and to explore the possible relationship between autophagy and apoptosis in order to provide a new direction for the study on the mechanism of infertility induced by varicocele . Methods Forty SD male rats were divided into normal control group, VC group (VC model group), VC group (VC + operation group) and sham operation group (n = 10). The normal control group without any treatment, VC group and VC surgery group modeling, sham operation group open the abdominal cavity without any treatment, on the 30th day VC surgery group left varicocele ligation surgery. On the 60th day, all the rats underwent left orchidectomy. The testicular tissue was stained with HE staining, TUNEL staining and transmission electron microscopy. The contents of LC3-Ⅱ and Beclin1 were detected by Western-blot and RT-PCR. Results Compared with normal control group and sham-operation group, the number of autophagosomes and autophagy lysosomes in the control group and the sham operation group was significantly lower than that in the VC group and VC group. The autophagosomes and autophagy lysosomes increased significantly in the VC and VC groups. Western-blot and RT-PCR showed that the expression of Beclin-1 and LC3-Ⅱ / Ⅰ in VC group and VC group was significantly higher than that in normal control group and sham operation group. Conclusion VC can induce the increase of autophagosome expression in rat testicular seminiferous tubule epithelial cells, and preliminary study the relationship between autophagy and apoptosis. The study on the autophagy of VC testicular cells may provide new targets and theoretical basis for the clinical treatment of VC.