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外源性神经激肽B对雌性大鼠生殖轴GnRH和Kisspeptin表达的影响
李海玲,司文宇,宋爽,康铁柱,姚郅璆,叶菁,李福宝,方富贵
0
(安徽农业大学 动物科技学院, 合肥 230036)
摘要:
为研究外源性神经激肽B(NKB)对雌性大鼠下丘脑促性腺激素释放激素(GnRH)和下丘脑-垂体-卵巢轴(HPOA)GnRH和Kisspeptin表达的影响,将40只60 d的Sprague-Dawley(SD)雌性大鼠随机分为试验组和对照组,侧脑室分别注射NKB溶液和等体积生理盐水,20 min后处死采集样品。采用免疫荧光技术分析各组织的GnRH和Kisspeptin蛋白表达,实时荧光定量PCR(RT-qPCR)分析GnRHKiss1mRNA的表达量变化。结果显示,与对照组相比,试验组下丘脑GnRH mRNA显著增加(P<0.05),Kiss1 mRNA表达量显著降低(P<0.05);试验组下丘脑视前区(POA)中GnRH阳性细胞数量显著增加(P<0.05),弓状核(ARC)和正中隆起(ME)中GnRH阳性细胞数量增加(P>0.05),但下丘脑ARC,POA,室旁核(PVN)和前腹侧室旁核(AVPV)中Kisspeptin阳性细胞数显著减少(P<0.05);且NKB抑制腺垂体中Kisspeptin的表达(P<0.05),但不影响卵巢中Kisspeptin的表达。综上,NKB通过调节HPOA Kisspeptin和下丘脑GnRH的表达来调控雌性大鼠的生殖。
关键词:  大鼠  神经激肽B  Kisspeptin  促性腺激素释放激素  下丘脑-垂体-卵巢轴
DOI:10.11841/j.issn.1007-4333.2021.01.08
投稿时间:2019-12-03
基金项目:国家自然科学基金项目(31972629)
Effects of exogenous neurokinin B on the expression of GnRH and Kisspeptin in the reproductive axis of adult female rats
LI Hailing,SI Wenyu,SONG Shuang,KANG Tiezhu,YAO Zhiqiu,YE Jing,LI Fubao,FANG Fugui
(College of Animal Sciences and Technology, Anhui Agricultural University, Hefei 230036, China)
Abstract:
To investigate the effect of exogenous neurokinin B(NKB)on the expression of gonadotropin releasing hormone(GnRH)in the hypothalamus and Kisspeptin in hypothalamic-pituitary-ovarian axis(HPOA)in female rats, forty Sprague-Dawley rats at the age of 60 d were divided into experimental group and control group randomly. Rats in the experimental group were injected with NKB solution into the lateral ventricle, and those in the control group were treated with equal volume of saline. All rats were sacrificed 20 min after the lateral ventricle treatment. Immunofluorescence method was used to analyze the expression of GnRH and Kisspeptin protein in each tissue. The expression levels of GnRH mRNA and Kiss1 mRNA in hypothalamus were analyzed by RT-qPCR. The results showed that: The expression of GnRH mRNA in the hypothalamus was significantly increased(P <0. 05)and Kiss1 mRNA was significantly decreased(P<0. 05)in experimental group compared with the control group; The number of GnRH-positive cells was significantly increased in preoptic area(POA)(P<0. 05)of the hypothalamus, and was increased in arcuate nucleus(ARC)and median eminence(ME)(P>0. 05), but a significantly decrease(P<0. 05)was found in the number of Kisspeptin-positive cells in ARC, POA, paraventricular nucleus(PVN)and anterior ventral paraventricular nucleus(AVPV)of the hypothalamus; NKB inhibited the expression of Kisspeptin in the anterior pituitary(P<0. 05), but did not affect the expression of Kisspeptin in the ovary. In summary, NKB regulates the reproduction of female rats by regulating the expression of Kisspeptin and GnRH on HPOA.
Key words:  rat  neurokinin B  Kisspeptin  gonadotropin releasing hormone  hypothalamic-pituitary-ovarian axis