神经营养因子受体Trkb对湖羊垂体细胞增殖及促性腺激素分泌的影响  

Effects of neurotrophic factor receptor Trkb on pituitary cell proliferation and gonadotropin secretion of Hu sheep

作　　者：陈培勇 蔡玉 杨花 徐辉 王锋 张艳丽 CHEN Peiyong;CAI Yu;YANG Hua;XU Hui;WANG Feng;ZHANG Yanli(Institute of Sheep and Goat Science,College of Animal Science and Technology,Nanjing Agricultural University,Nanjing 210095,China)

机构地区：[1]南京农业大学动物科技学院羊业科学研究所,江苏南京210095

出　　处：《南京农业大学学报》2024年第1期166-173,共8页Journal of Nanjing Agricultural University

基　　金：国家自然科学基金项目(32072737);江苏现代农业(肉羊)产业体系项目(060/61KA210031)。

摘　　要：[目的]本研究旨在探究神经营养因子酪氨酸激酶B受体(Trkb)基因对湖羊垂体促性腺激素分泌的影响。[方法]利用qPCR方法对Trkb进行组织表达谱分析;构建Trkb过表达载体并转染至湖羊垂体细胞,利用qPCR、Western blot、EdU以及ELISA等技术检测过表达Trkb对垂体细胞增殖及促性腺激素分泌的影响。[结果]Trkb在湖羊心、肝、脾、肺、肾以及下丘脑和垂体等各个组织中均有表达,但在垂体中表达水平显著高于其他组织(P<0.05)。Trkb在湖羊垂体组织不同发育阶段差异表达,其中在6月龄垂体组织中高表达(P<0.05),在5日龄和3月龄表达水平较低。与对照组相比,过表达Trkb基因显著促进了垂体细胞增殖率(P<0.05),增殖标记基因Pcna表达水平与Bcl2/Bax比值均显著提高(P<0.05)。此外,过表达Trkb显著提高了促性腺激素相关基因Fshβ和Lhβ的表达水平,促进了垂体细胞促卵泡素(FSH)分泌(P<0.05)。[结论]过表达Trkb能够显著促进湖羊垂体细胞增殖,降低细胞凋亡水平从而显著提高促性腺激素的分泌水平。本研究初步验证Trkb基因在湖羊垂体细胞中功能,为深入研究Trkb调控垂体功能的分子机制提供了试验依据。[Objectives]The study aimed to investigate the neurotrophic factor tyrosine kinase B receptor(Trkb)gene effects on the pituitary gonadotropin secretion of Hu sheep.[Methods]Tissue expression profiling of Trkb was performed using qPCR.The overexpression vector of Trkb was constructed and transfected into Hu sheep pituitary cells.The effects of overexpression of Trkb on pituitary cell proliferation and gonadotropin secretion were detected by qPCR,Western blot,EdU and ELISA.[Results]Trkb was expressed in the heart,liver,spleen,lung,kidney,hypothalamus,pituitary and other tissues of Hu sheep,but the expression level in the pituitary was significantly higher than that in other tissues(P<0.05).Trkb was differentially expressed in pituitary tissues of Hu sheep at different developmental stages,and was highly expressed in pituitary tissues at 6 months of age(P<0.05),and the expression level was lower at 5 days and 3 months of age.Compared with the control group,overexpression of Trkb gene significantly promoted pituitary cell proliferation rate(P<0.05),and the expression levels of proliferation marker gene Pcna and Bcl2/Bax ratio significantly increased(P<0.05).In addition,overexpression of Trkb significantly increased the expression levels of gonadotropin-related genes Fshβ and Lhβ,and significantly promoted the secretion levels of follicle stimulating hormone(FSH)in pituitary cells(P<0.05).[Conclusions]Overexpression of Trkb could significantly promote cell proliferation efficiency of pituitary cells,decrease the level of cell apoptosis,and thus significantly increase the secretion of gonadotropin in Hu sheep.This study preliminarily verified the function of Trkb in Hu sheep pituitary cells and provided a experimental basis for further study of the molecular mechanism of Trkb regulating pituitary function.

关 键 词：湖羊 垂体 酪氨酸激酶B受体(Trkb) 细胞增殖 促性腺激素 

分 类 号：S852.6[农业科学—基础兽医学]