黄精不同部位改善“肾精亏虚”雄鼠生殖功能低下的作用研究

Effects of different fractions of Polygonati Rhizoma on reproductive dysfunction in male mice with kidney essence deficiency

研究黄精不同部位对房事不节致肾精亏虚模型雄鼠生殖功能低下的作用与机制。选取性功能良好的雄性ICR小鼠50只,随机分为正常组(NC)、模型组(MC)、黄精正丁醇部位组(0.4 g·kg^(-1), HJCT)、黄精剩余部位组(0.4 g·kg^(-1), HJSY)、黄精超微粉组(1.0 g·kg^(-1), HJCW)。采用Coolidge效应诱导雄性小鼠房事不节,建立肾精亏虚生殖功能低下小鼠模型。造模同时给予相应药物,连续8周。测定小鼠肛温、抓力、自主活动次数、负重游泳时间、性行为(捕捉、骑跨和射精)等肾精亏虚证候指标;采用ELISA法测定血清中睾酮(T)、促黄体生成素(LH)、精子生成素(FSH)、皮质醇(COR)、促性腺激素释放激素(GnRH)、雌二醇(E_(2))等激素水平及精子内精浆果糖含量;采用苏木素-伊红(HE)染色观察睾丸组织病理变化;检测精子质量(精子密度、精子活力、精子存活率以及精子畸形率);采用Western blot法检测睾丸组织增殖及凋亡蛋白表达。黄精正丁醇部位、剩余部位以及超微粉能显著增加模型小鼠抓力、肛温、自主活动次数、负重游泳时间以及性行为(捕捉、骑跨以及射精)次数,升高血清中T、LH、GnRH、E_(2)激素含量,降低COR和FSH水平激素含量;增加睾丸组织的湿重;增加模型小鼠精子密度、精子活力以及精子存活率,降低精子畸形率;升高模型小鼠精囊中的精浆果糖的含量;减轻小鼠睾丸病变,抑制睾丸组织细胞的凋亡。黄精正丁醇部位改善下丘脑-垂体-性腺轴(HPGA)紊乱,增强性行为以及改善精子质量方面优于黄精剩余部位和黄精超微粉;黄精超微粉抑制睾丸组织细胞凋亡方面优于黄精正丁醇部位和黄精剩余部位。黄精各部位均能够改善“肾精亏虚”雄鼠生殖功能,抑制睾丸组织细胞凋亡情况,其作用机制可能与改善HPGA紊乱有关。

This study aims to reveal the effects and mechanisms of different fractions of Polygonati Rhizoma on the reproductive dysfunction in male mice with kidney essence deficiency due to excess of sexual intercourse.Fifty male ICR mice with good sexual function were selected and randomized into normal(NC),model(MC),n-butanol fraction of Polygonati Rhizoma(0.4 g·kg^(-1),HJCT),remaining fraction of Polygonati Rhizoma(0.4 g·kg^(-1),HJSY),and ultra-micro powder of Polygonati Rhizoma(1.0 g·kg^(-1),HJCW)groups.The Coolidge effect was used to induce excess of sexual intercourse in the mice,and a mouse model of reproductive dysfunction with kidney essence deficiency was established.The mice were administrated with corresponding drugs at the same time of modeling for 8 consecutive weeks.The anal temperature,grip strength,locomotor activities,weight-bearing swimming time,and sexual behaviors(catching,straddling,and ejaculation)were measured.Enzyme-linked immunosorbent assay was employed to measure the levels of testosterone(T),luteinizing hormone(LH),follicle-stimulating hormone(FSH),cortisol(COR),gonadotropin-releasing hormone(GnRH),and estradiol(E_(2))in the serum and the content of fructose in seminal plasma.Hematoxylin-eosin(HE)staining was employed to observe the pathological changes in the testicular tissue.The sperm quality was evaluated based on the density,viability,survival rate,and deformation rate of sperms.Western blot was employed to determine the expression of proteins involved in proliferation and apoptosis in the testicular tissue.The results showed that HJCT,HJSY,and HJCW significantly increased the grip strength,anal temperature,locomotor activities,weight-bearing swimming time,and sexual behaviors,elevated the serum levels of T,LH,GnRH,and E_(2),lowered the levels of COR and FSH,and increased the wet weight of the testicular tissue in the model mice.In addition,they increased the density,viability,and survival and reduced the deformation rate of sperms,increased the content of fructose in the seminal plasma,alleviated the pathological changes and inhibited apoptosis in the testicular tissue in the model mice.HJCT was superior to HJSY and HJCW in ameliorating the hypothalamic-pituitary-gonadal axis(HPGA)disorder,enhancing sexual behaviors,and improving sperm quality.HJCW outperformed HJCT and HJSY in inhibiting apoptosis in the testicular tissue.In conclusion,HJCT,HJSY,and HJCW can improve the reproductive function and inhibit apoptosis in the testicular tissue of the male mice with kidney essence deficiency by ameliorating the HPGA disorder.