摘要
目的研究人工光环境对雌性豚鼠生殖内分泌的影响及日钟阴阳方对雌性豚鼠下丘脑中吻素Kisspeptin及吻素受体(KISS1R)的作用机制。方法将3月龄雌性豚鼠按体重的大小从1到36编号,获取随机数字,按余数分为6组,分别为自然光组、自然光食疗组、LED灯组、LED灯食疗组、荧光灯组和荧光灯食疗组,每组6只。用日钟阴阳方对自然光食疗组、LED灯食疗组、荧光灯食疗组进行干预12周。饲养12周后,于动情前期取材,HE染色,电镜下观察卵巢形态;ELISA测定豚鼠血清中雌二醇(E2)、促黄体生成素(LH)、促卵泡生成素(FSH)、睾酮(T)、抗苗勒氏管激素(AMH)、泌乳素(PRL)水平;Western Blot测定豚鼠卵巢内雄激素受体(AR)、促黄体生成素受体(LHR)、促卵泡生成素受体(FSHR)及下丘脑中KISS1R的蛋白含量及逆转录聚合酶链式反应法(RT-PCR)测定豚鼠卵巢内AR、LHR、FSHR mRNA及下丘脑中KISS1R mRNA表达水平。结果(1)自然光照组及自然光食疗组可见结构完整的卵母细胞,未见明显病理变化;荧光灯组及LED灯组卵巢卵母细胞消失,见大量炎性细胞浸润;荧光灯食疗组及LED灯食疗组卵巢可见卵母细胞,但卵母细胞周围粒细胞排列疏松,间隙增大。(2)与自然光照组比较,自然光食疗组血清E2、LH、FSH、T、AMH、PRL含量下降,下丘脑内KISS1R及卵巢内LHR、FSHR、AR蛋白含量下降(P<0.05);与荧光灯组比较,荧光灯食疗组血清血清E2、LH、FSH、T、AMH、PRL含量下降,下丘脑内KISS1R及卵巢内LHR、FSHR、AR蛋白含量下降(P<0.05);与LED灯组比较,LED灯食疗组血清E2、LH、FSH、T、AMH、PRL含量均下降,下丘脑内KISS1R及卵巢内LHR、FSHR、AR蛋白含量下降(P<0.05);与自然光照组比较,荧光灯、LED灯组血清E2、LH、FSH、T、AMH、PRL含量上升,下丘脑内KISS1R及卵巢内LHR、FSHR、AR蛋白含量上升(P<0.05);与自然光食疗组比较,荧光灯食疗、LED灯食疗组血清E2、LH、FSH、T、AMH、PRL含量上升,下丘脑内KISS1R及卵巢内LHR、FSHR、AR蛋白含量上升(P<0.05)。结论人工光环境的改变会对豚鼠生殖内分泌造成影响,日钟阴阳方可有效改善豚鼠生殖内分泌紊乱的结局。其可能机制与通过调控豚鼠下丘脑内KISS1R的表达,影响Kisspeptin的分泌有关。
Objective To study the effect of artificial light environment on reproductive endocrine of female guinea pigs and to observe the intervention effect of Rizhong Yinyang Formula(RZYYF)on Kisspeptin and its receptor KISS1R in hypothalamus of female guinea pigs.Methods The 3-month-old female guinea pigs were numbered from 1 to 36 according to their body weight,and the random numbers were obtained.They were divided into 6 groups according to the remainder:natural light group,natural light diet group,LED light group,LED light diet group,fluorescent light group,and fluorescent light diet group,6 guinea pigs in each group.The natural light diet group,LED light diet group,and fluorescent light diet group were intervened for 12 weeks.After feeding for 12 weeks,the materials were taken in the pre estrus period,stained with HE,and the morphology of ovary was observed under electron microscope.The hormone levels of estradiol(E_(2)),luteinizing hormone(LH),follicle stimulating hormone(FSH),testosterone(T),anti-Mϋllerian hormone(AMH)and prolactin(PRL)in guinea pig serum were measured by ELISA.The protein contents of androgen receptor(AR),luteinizing hormone receptor(LHR),follicle stimulating hormone receptor(FSHR)in guinea pig ovary and KISS1R in hypothalamus were measured by Western Blot,and mRNA expression levels of AR,LHR,FSHR in guinea pig ovary and KISS1R mRNA in hypothalamus were measured by reverse transcription polymerase chain reaction(RT-PCR).Results(1)Oocytes with complete structure were observed in natural light group and natural light diet group,and no obvious pathological changes were found.In the fluorescent lamp group and LED lamp group,ovarian oocytes disappeared and a large number of infiltrated inflammatory cells were seen.Oocytes could be seen in the ovaries of the two groups,but the arrangement of granulocytes around the oocytes was loose and the gap was enlarged.(2)Compared with the natural light group,the contents of serum E_(2),LH,FSH,T,AMH,and PRL in the natural light diet group decreased,and the protein contents of KISS1R in hypothalamus and LHR,FSHR and AR in ovary decreased(P<0.05).Compared with the fluorescent lamp group,the contents of serum E_(2),LH,FSH,T,AMH,and PRL in the fluorescent lamp diet group decreased,and the protein contents of KISS1R in hypothalamus and LHR,FSHR,and AR in ovary decreased(P<0.05).Compared with the LED light group,the contents of serum E_(2),LH,FSH,T,AMH,and PRL in the LED light diet group decreased,and the protein contents of KISS1R in hypothalamus and LHR,FSHR and AR in ovary decreased,except AMH in the LED light diet group(P<0.05).Compared with natural light group,the contents of serum E_(2),LH,FSH,T,AMH,and PRL in fluorescent lamp and LED lamp group increased,and the contents of KISS1R in hypothalamus and LHR,FSHR and AR in ovary increased(P<0.05).Compared with the natural light diet group,the contents of serum E_(2),LH,FSH,T,AMH,and PRL in the fluorescent light diet group and LED light diet group increased,and the contents of KISS1R in hypothalamus and LHR,FSHR and AR protein in ovary increased in the fluorescent light diet group and LED light diet group(P<0.05).Conclusions The changes of artificial light environment will affect the reproductive endocrine of guinea pigs.RZYYF effectively improved the outcomes of reproductive endocrine disorders of guinea pigs.The possible mechanism might be through regulating the expression of KISS1R in the hypothalamus of guinea pigs,which might be related to regulating KISS1R expression in hypothalamus of guinea pigs and affecting the secretion of Kisspeptin.
作者
施加裕
谈勇
SHI Jia-yu;TAN Yong(First Clinical Medical College,Nanjing University of Traditional Chinese Medicine,Nanjing,210029;Department of Reproductive Medicine,Affiliated Hospital,Nanjing University of Traditional Chinese Medicine,Nanjing,210001)
出处
《中国中西医结合杂志》
CAS
CSCD
北大核心
2021年第12期1468-1475,共8页
Chinese Journal of Integrated Traditional and Western Medicine
基金
国家自然科学基金面上项目(No.81873333,No.81674012)
江苏省科教强卫工程项目(No.YXZXB2016006)。
关键词
光环境
日钟阴阳方
性激素水平
吻素
吻素受体
light environment
Rizhong Yinyang Formula
sex hormone level
Kisspeptin
Kisspeptin receptor
