菟丝子黄酮干预雷公藤多苷所致雄性幼鼠生殖损伤

目的:研究不同剂量菟丝子黄酮干预雷公藤多苷(GTW)所致雄性幼鼠生殖损伤的影响。方法:雄性SD幼鼠80只,分2批喂养,每批40只。每批随机分为5组,空白组ig羧甲基纤维素钠(CMC-Na)],多苷组(ig GTW,9 mg·kg~(-1)·d~(-1)),黄酮高、中、低剂量组(ig菟丝子+GTW,0.2 g·kg~(-1)+9 mg·kg~(-1),0.1 g·kg~(-1)+9 mg·kg~(-1),0.05 g·kg~(-1)+9 mg·kg~(-1)),持续给药4,12周,分别在停药时麻醉处死留取睾丸组织,称重检测脏器指数,苏木素伊红(HE)染色观察睾丸组织病理变化,实时荧光定量PCR(Real-time PCR)及免疫印迹法(Western blot)检测雄激素受体(AR)mRNA及蛋白表达。结果:给药4,12周时多苷组大鼠体重、睾丸质量较空白组显著降低(P0.01);黄酮组大鼠体重、睾丸质量较多苷组显著提高(P0.01);黄酮中剂量组大鼠体重、睾丸质量较黄酮高、低剂组明显增高(P0.05)。给药4周时,多苷组大鼠睾丸曲细精管间距变宽、曲细精管扭曲、水肿,严重者曲细精管破裂,间质细胞散乱,精原细胞、精母细胞明显减少,12周时损伤程度更重;黄酮组中剂量上述病理损伤较轻。给药4,12周后,多苷组大鼠睾丸组织AR mRNA及蛋白表达明显下调(P0.05,P0.01),黄酮中剂量可以明显提高大鼠睾丸组织AR mRNA及蛋白表达(P0.05,P0.01)。结论:GTW可以明显降低大鼠体重、睾丸质量;下调睾丸组织AR mRNA及蛋白水平表达;GTW对雄性大鼠睾丸组织有明显的损伤作用,且随剂量的增加和时间延长损伤程度越重;菟丝子黄酮中剂量组保护这种损伤作用更加显著。

Effect of Different Doses of Cuscutae Semen Flavone on Tripterygium Glycosides-induced Reproductive Damage in Male Juvenile Rats

Objective: To investigate the effects of different doses of Cuscutae Semen flavone on glycosides of Tripterygium wilfordii (GTW)-induced reproductive damage in male juvenile rats. Method: The 80 3-4 week-old SD male rats were randomly divided into 2 batches for feeding, n=40 in each batch. The rats in each batch were further randomly divided into 5 groups, n=8 in each group:blank group(ig CMC-Na), GTW (ig 9 mg·kg^-1·d^-1) group, Cuscutae Semen flavone groups (high dose group:0.2 g·kg^-1·d^-1+GTW 9 mg·kg^-1·d^-1, middle dose group:0.1 g·kg^-1·d^-1+GTW 9 mg·kg^-1·d^-1, low dose group:0.05 g·kg^-1·d^-1+GTW 9 mg·kg^-1·d^-1). In the flavone groups, GTW was mixed with flavone into rats by intragastric administration for continuous 4 weeks and 12 weeks. After treatment, rats were killed under anesthesia and testicular tissues were taken. The body weight of the rats was observed and organ indexes of testicular tissue specimens were detected. The pathological changes of testicular tissues were observed by HE staining method, and the androgen receptor (AR) mRNA and protein expression levels were detected by Real-time PCR and Western blot. Result: GTW can significantly reduce body weight and testicular weight after 4 weeks and 12 weeks of treatment. Compared with the blank group, the body weight and testicular weight of rats in GTW group were significantly lower (P < 0.01), compared with GTW group, the body weight and testicular weight of rats in flavone groups were significantly increased (P < 0.01), the body weight and testicular weight of rats in flavone middle dose group were significantly higher than those in high dose group and low dose group (P < 0.05).After treatment for 4 weeks, space of convoluted tubule in testis of rats in GTW group was broadened, convoluted tubule was distorted with edema, in severe cases, convoluted tubule was broken with scattered mesenchymal cells and significantly reduced spermatogonia and spermatocyte, the testicular damage was heavier at week 12. The above pathological damage was lighter in flavone middle dose group. After 4 weeks administration, AR mRNA and protein expression levels in testicular tissues of rats in GTW group were significantly down-regulated (P < 0.05), and after 12 weeks administration, AR mRNA and protein expression levels were decreased more significantly (P < 0.01). Flavone group (middle dose) can significantly increase the AR mRNA and protein expression levels in testicular tissues (P < 0.05, P < 0.01). Conclusion: GTW can significantly reduce the body weight and testicular weight of rats, down-regulate AR mRNA and protein expression levels, GTW had significant damaging effect to the testicular tissues of male rats in a dose-dependent and time-dependent manner, Cuscutae Semen flavone middle dose group had a more significant protective effect for such damages.