肉苁蓉苯乙醇苷对环磷酰胺致小鼠生精障碍的治疗作用及其机制

摘要

目的：探讨肉苁蓉苯乙醇苷对环磷酰胺致生精障碍小鼠的治疗作用，并初步阐明其作用机制。方法：乙醇浸提法提取肉苁蓉苯乙醇苷。40只BALB／C小鼠随机分为对照组、模型组、肉苁蓉苯乙醇苷低剂量组（50 mg·kg-1）和高剂量组（100 mg· kg-1），每组10只。除对照组外，其余各组小鼠每天注射环磷酰胺（80 mg·kg-1）连续5 d，制备生精障碍小鼠模型。末次给药后，肉苁蓉苯乙醇苷低剂量组和高剂量组小鼠分别给予相应剂量灌胃，对照组和模型组小鼠给予等体积生理盐水灌胃，每天1次，连续30 d。末次灌胃后24 h，取小鼠睾丸组织。酶联免疫法测定小鼠睾丸组织匀浆中睾酮水平，比较各组小鼠精子密度、精子活率及精子畸形率， HE染色观察睾丸组织形态学变化。结果：与对照组比较，模型组小鼠精子密度和精子活率降低（P＜0.01），精子畸形率升高（P＜0.01），睾丸组织匀浆中睾酮水平降低（P＜0.01）；与模型组比较，肉苁蓉苯乙醇苷各剂量组小鼠精子密度和精子活率明显升高（P＜0.01），精子畸形率降低（P＜0.01），睾丸组织匀浆中睾酮水平升高（P＜0.05或P＜0.01）。组织学观察，模型组小鼠睾丸生精小管萎缩、退化，生精上皮明显变薄、生精细胞排列紊乱，腔内精子数目减少；肉苁蓉苯乙醇苷组小鼠睾丸生精上皮层数明显增多，层次分明，生精细胞排列整齐、紧密、规则，管腔内可见精子。结论：肉苁蓉苯乙醇苷对环磷酰胺所致小鼠生精障碍具有明显的治疗作用，其机制可能与改善睾丸组织中睾酮水平有关。%Objective To explore the therapeutic effect of phenylethanoid glycosides on cyclophosphamide (CTX)-induced dyszoospermia in mice and to preliminary elucidate the mechanisms involved in the process. Methods Phenylethanoid glycoside was extracted by ethanol extraction.Forty male BALB/C mice were randomly divided into control group,model group,low dose of phenylethanoid glycosides group (50 mg· kg-1 )and high dose of phenylethanoid glycosides group (100 mg·kg-1 ).Except control group,the dyszoospermia mouse model was established by peritoneal injection of CTX at the daily dose of 80 mg· kg-1 ,once daily for successive 5 d. After modeling, phenylethanoid glycosides were intragastrically administered at corresponding doses to each phenylethanoid glycosides group.Equal volume of normal saline was given to the mice in control group and model group by gastrogavage.All the medication was performed once daily for successive 30 d.The testis tissue was obtained 24 h after the last intragastric administration.The level of testosterone in the testis tissue homogenate was determined by enzyme linked immunosorbent assay.The sperm counts, the motility rates, and the teratospermia rates in various groups were compared.The morphological changes of the testis tissue were observed using HE staining.Results Compared with control group, the sperm count and the motility rate were decreased, the teratospermia rate was increased,and the testosterone level in the testis tissue homogenate was decreased in model group(P<0.01).Compared with model group,the sperm counts and the motility rates were increased,the teratospermia rates were decreased, and the testosterore levels in the testis tissue homogenate were increased in phenylethanoid glycosides groups (P<0.05 or P<0.01).The histological results showed atrophy and degeneration of seminiferous tuble,thicker seminiferous epithelium and azoospermic lumina in model group;the number of seminiferous epithelial layers was increased and the seminiferous cells orderly arranged, and many sperms were found in the tubules in phenylethanoid glycosides groups.Conclusion Phenylethanoid glycosides has obviously therapeutical effect on CTX-induced dyszoospermia in mice,and its mechanisms might be correlated with recovering the testosterone level.