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目的:环境因素及抗癌药物的使用会导致生精细胞出现凋亡,从而引起男性不育。目前尚缺乏疗效明确的治疗药物。左卡尼汀被用来治疗男性不育取得了一定的疗效。本研究拟通过环磷酰胺诱导的大鼠不育模型,研究左卡尼汀对生精细胞的保护作用。创新点:首次观察了左卡尼汀改善化疗药物环磷酰胺所导致的生殖功能障碍的疗效。方法:采用环磷酰胺诱导大鼠不育模型成功后,给予左卡尼汀治疗15天,然后处死全部大鼠。检测血清中的睾酮和雌激素含量;采用全自动精子分析仪检测附睾组织中精子的数量和质量;采用苏木精-伊红(H&E)染色观察睾丸组织的病理损伤;通过末端脱氧核苷酸转移酶介导的d UTP缺口标记技术(TUNEL)的染色观察睾丸组织中生精细胞的凋亡;采用免疫组织化学法观察睾丸组织中LC3和Beclin-1的表达;采用蛋白质免疫印迹分析(Western blot)和多聚酶链式反应(PCR)分别检测组织中LC3和Beclin-1的表达。结论:左卡尼汀可以通过增加细胞自噬,抑制生精细胞凋亡,提高生精细胞的质量和数量,进而降低环磷酰胺引起的睾丸组织的损伤。
Purpose: Environmental factors and the use of anti-cancer drugs can lead to apoptosis of spermatogenic cells, causing male infertility. Currently there is a lack of effective treatment of drugs. L-carnitine has been used to treat male infertility achieved some effect. In this study, cyclophosphamide-induced rat infertility model to study the protective effect of levocarnitine on spermatogenic cells. Innovation: For the first time observed the efficacy of levocarnitine to improve the reproductive dysfunction caused by cyclophosphamide chemotherapy. Methods: After successful treatment of cyclophosphamide-induced rat model of infertility, L-carnitine was given for 15 days and all the rats were sacrificed. The quantity and quality of spermatozoa in the epididymis were detected by automatic sperm analyzer. The pathological damage of testis was observed by hematoxylin and eosin (H & E) staining. The expression of testosterone and estrogen in the testis was detected by terminal deoxynucleotide The apoptosis of spermatogenic cells in testis was observed by TUNEL staining. The expression of LC3 and Beclin-1 in testis was observed by immunohistochemical staining. Western blotting analysis blot and polymerase chain reaction (PCR) were used to detect the expression of LC3 and Beclin-1 respectively. CONCLUSION: L-carnitine can decrease the apoptosis of testicular tissue induced by cyclophosphamide by increasing autophagy, inhibiting the apoptosis of spermatogenic cells, improving the quality and quantity of spermatogenic cells.