摘要
目的观察AT1受体对雄性小鼠生殖系统的调控作用,初步探讨中药复方五子衍宗丸改善♂AT1+/-小鼠生殖功能的作用机制。方法选择8周龄C57BL/6正常雄性小鼠20只,♂AT1+/-小鼠40只,随机分为A组(正常雄性小鼠)、B组(♂AT1+/-小鼠组)、C组(♂AT1+/-小鼠+五子衍宗丸组)。A组和B组予以0.9%NaC l溶液灌胃,C组予以五子衍宗丸(5.8 g/kg)灌胃,每日2次,每次0.5 mL。采用RT-PCR法测定睾丸、下丘脑、垂体各组AT1表达,取样前行受孕率测定,并对各组精子进行形态观察,ELISA酶免法测定各组血浆性激素水平,包括促性腺激素释放激素(GnR H)、促卵泡生成素(FSH)、黄体生成素(LH)和睾酮(T)。结果在睾丸、垂体、下丘脑组织中,B组和C组仅检测出少量AT1,与正常组比较,差异有统计学意义(P<0.01),说明模型成功;与B组和C组合笼的雌鼠无一受孕;B组和C组精子形态明显异常,与正常组比较,差异有统计学意义(P<0.01),经过五子衍宗丸干预后,精子正常率明显提高,C组与B组差异有统计学意义(P<0.05)。B组GnR H水平与A组差异有统计学意义(P<0.01)。经过五子衍宗丸干预后,C组GnR H、FSH明显提高,与B组差异有统计学意义(P<0.05);C组第4周与同组第2周相比,GnR H、FSH、LH水平均有提高(P<0.05)。结论AT1基因敲除可导致雄性小鼠不育,五子衍宗丸可能通过提高♂AT1+/-小鼠的性激素水平,提高精子正常率,从而改善雄性小鼠生殖功能。
Objective To observe the regulation effect of AT1 receptor on the reproductive system of male mice,and preliminarily explore the mechanism of improving the reproductive function of♂AT1+/-mice by Wuzi Yanzong pills,a traditional Chinese medicine compound.Methods Twenty 8-week-old C57 BL/6 normal male mice and 40 AT1 KO heterozygous male mice(♂AT1+/-)were randomly divided into group A(normal male mice group),group B(♂AT1+/-mice group),and group C(♂AT1+/-mice+Wuzi Yanzong pills group).The mice in group A and group B were given intragastric administration with 0.9%NaC l solution,and the mice in group C with Wuzi Yanzong pills at 5.8 g/kg,twice daily with 0.5 mL each time.The expression of AT1 in testis,hypothalamus and pituitary was determined by RT-PCR method.Pregnancy rate was measured before sampling,and sperm morphology was observed in each group.Plasma sex hormone levels,including gonadotropin releasing hormone(Gn RH),follicle stimulating hormone(FSH),luteinizing hormone(LH)and testosterone(T),were measured by ELISA enzyme immunoassay.Results Compared with the normal group,only a small amount of AT1 was detected in testis,pituitary and hypothalamus tissues in both groups of B and C(P<0.01),indicating that the model was successful.None of the female mice in cages with mice from group B and C were pregnant,but the sperm morphology in group B and C was significantly abnormal as compared with that in the normal group(P<0.01).After the intervention of Wuzi Yanzong pills,the normal rate of sperm was significantly increased,and there was significant difference between group C and group B(P<0.05).The level of Gn RH in group B was significantly higher than that in group A(P<0.01).After the intervention of Wuzi Yanzong pills,the levels of Gn RH and FSH in group C were significantly higher than those in group B(P<0.05).The levels of GnRH,FSH and LH in group C at the 4 th week were higher than those in the second week of the same group(P<0.05).Conclusion The knockout of AT1 gene can lead to infertility in male mice.
作者
邹贇
赵方方
李慧
张亚亨
赵雪昊
刘雪乐
吴欣怡
杨雪军
ZOU Yun;ZHAO Fangfang;LI Hui;ZHANG Yaheng;ZHAO Xuehao;LIU Xuele;WU Xinyi;YANG Xuejun(Department of Nephrology,Shuguang Hospital Affiliated to Shanghai University of Traditional Chinese Medicine,Shanghai 201203,China)
出处
《上海中医药杂志》
2020年第6期97-100,共4页
Shanghai Journal of Traditional Chinese Medicine
基金
国家自然科学基金项目(81202669)
关键词
肾素-血管紧张素系统
不育
小鼠
肾主生殖
五子衍宗丸
renin-angiotensin system
infertility
mice
kidney governing reproduction
Wuzi Yanzong pills