“菟丝子-枸杞子”药对修复生精功能障碍大鼠血睾屏障的机制研究

目的观察"菟丝子-枸杞子"药对对雷公藤多苷(GTW)诱导的生精障碍大鼠模型血睾屏障的影响,初步探讨其改善模型大鼠生殖功能的机理。方法将56只SD大鼠随机分为空白对照组、模型对照组、阳性对照组和药对组。以GTW 40mg/(kg·d)灌胃4周构建SD大鼠生精障碍模型,成模后阳性对照组每日给予左卡尼汀悬液0.2g/(kg·d)]灌胃,药对组每日给予菟丝子-枸杞子悬液6g/(kg·d)]灌胃,治疗4周后检测各组大鼠的精子质量、生殖激素和抑制素B、附睾肉毒碱,观察支持细胞形态,并采用Western blot检测睾丸组织血睾屏障相关蛋白的表达,分析比较不同组间的差异。结果药对组大鼠精子浓度(39.17±13.75)×10~6/mL]高于模型对照组(28.23±8.05)×10~6/mL],其差异具有统计学意义(P<0.05)。药对组(29.92±5.25)%]和左卡尼汀组精子活动率(29.31±6.33)%]均显著高于模型对照组大鼠(20.13±6.63)%],其差异具有统计学意义(P<0.01);药对组和左卡尼汀组大鼠卵泡生成素(FSH)、黄体生成素(LH)和泌乳素(PRL)与模型对照组相比均显著降低,附睾肉毒碱含量显著升高,其差异具有统计学意义(P<0.05或P<0.01)。药对组与模型对照组比较,支持细胞结构和形态改善,生精小管结构、排列较为正常,生精细胞明显增多,形态改善;药对组Occludin、ZO-1和connexin 43的蛋白表达比模型对照组显著增多,其差异具有统计学意义(P<0.05);β-catenin和N-cadherin的蛋白表达也有不同程度增高,虽然未见明显差异,但增高趋势明显。结论 "菟丝子-枸杞子"药对能够有效减轻GTW造成的生精功能障碍,其机制可能与修复血睾屏障损伤有关。

Study on the mechanism of semen cuscutae and fructus lycii repairing blood-testis barrier in rats with spermatogenic dysfunction

Objective To elucidate the effect of Semen Cuscutae-Fructus Lycii(SC-FL)on repairing blood-testis barrier induced by Glucosides of Tripterygium wilfordii hook.F(GTW),using a dyszoospermia rat model.Methods 56 sprague-dauley(SD)rats were randomly divided into four groups:normal control(treated with saline),GTW(treated with saline),GTW+levocarnitine(treated with levocarnitine),and GTW+SCFL(treated with SC-FL).GTW(40 mg/kg/d)was intragastrically administered to establish dyszoospermia models.After 4 weeks of treatment,semen quality,sex hormone,inhibin B,and L-carnitine levels were measured.Testicular tissue was observed using hematoxylin-eosin(H&E)staining.Western blotting was used to determine the expression of blood-testis barrier associated protein in rat testicular tissue.Results Compared with the GTW group(28.23±8.05)×10~6/mL],sperm concentration was significantly higher in the GTW+SCFL group(39.17±13.75)×10~6/mL](P<0.05).Sperm motility was much better in the GTW+SCFL(29.92±5.25)%]and GTW+levocarnitine group(29.31±6.33)%]than that in the GTW group(20.13±6.63)%](P<0.01).Compared with the GTW group,follicle stimulating hormone(FSH),luteinizing hormone(LH)and prolactin(PRL)were significantly lower in the GTW+SCFL and GTW+levocarnitine group,and L-carnitine levels was significantly higher in the two groups(P<0.05 or P<0.01).Histomorphology showed that in the GTW+SCFL group,the morphology of Sertoli cells and spermatogenic cells improved,the structure and arrangement of seminiferous tubules were better,and the amount of spermatogenic cells increased significantly,compared to those in the GTW group.Western blotting results showed that compared with the GTW group,the expression of occludin,ZO-1,and connexin 43 in the GTW+SCFL group was increased(P<0.05).The expression ofβ-catenin and N-cadherin was higher than those of the GTW group,although not statistically significant.Conclusions SC-FL can effectively improve the spermatogenic function of rats with spermatogenic dysfunction,and the mechanism may be related to repairing blood-testis barrier.