瘦素对绵羊卵泡颗粒细胞增殖/凋亡的影响  

Effect of Leptin on the Proliferation/Apoptosis of Sheep Follicular Granulosa Cells

作　　者：贾凯琪 郭翔宇 党文庆[1] 李雅婷 马晓燕 王锴[1] 吕丽华 JIA Kaiqi;GUO Xiangyu;DANG Wenqing;LI Yating;MA Xiaoyan;WANG Kai;Lü Lihua(College of Animal Science,Shanxi Agricultural University,Taigu 030801,China)

机构地区：[1]山西农业大学动物科学学院,山西太谷030801

出　　处：《山西农业科学》2021年第8期999-1005,共7页Journal of Shanxi Agricultural Sciences

基　　金：山西省青年三晋学者专项(J241644001)。

摘　　要：为了研究不同浓度瘦素对绵羊卵泡颗粒细胞的增殖凋亡作用及其作用机制,在发情期采集成年健康绵羊的双侧卵巢,收集卵泡颗粒细胞(GCs)进行培养,通过免疫荧光方法检测和确认所分离的细胞是卵泡颗粒细胞,通过EdU检测添加不同质量浓度瘦素(0、10、25、50、100 ng/mL)对细胞的增殖作用,通过qPCR、Western blotting检测瘦素对卵泡颗粒细胞的作用以及是否通过JAK2/STAT3通路发挥作用。结果表明,添加不同浓度瘦素处理GCs 24 h,添加25、50 ng/mL瘦素可促进颗粒细胞增殖;瘦素处理后能增强LEPR、JAK2、Bcl-2基因和LEPR、JAK2、P-STAT3、Bcl-2蛋白的表达,降低BAX基因的表达。不同浓度瘦素对绵羊卵泡颗粒细胞有增殖作用,50 ng/mL作用最佳;瘦素通过与受体相互作用激活JAK2/STAT3通路对绵羊卵泡颗粒细胞发挥抗凋亡作用,可为治疗人类和家畜代谢紊乱导致的低生育力或者不育提供新思路。To study the effects of different concentrations of leptin on the proliferation and apoptosis of sheep follicular granulosa cells and its mechanism,bilateral ovaries of healthy and mature sheep were collected during estrus,and granulosa cells(GCs)were collected for culture.The isolated cells detected and confirmed by immunofluorescence method were follicular granulosa cells.EdU was used to detect the effects of different concentrations of leptin(0,10,25,50,100 ng/mL)on cell proliferation.The effect of leptin on follicular granulosa cells and whether it played a role through the JAK2/STAT3 pathway were detected by qPCR and Western blotting.The results showed that different concentrations of leptin were added to GCs for 24 h,25 ng/mL leptin and 50 ng/mL leptin promoted the proliferation of granulosa cells.Leptin could increase the expression of LEPR,JAK2,Bcl-2 gene and LEPR,JAK2,P-STAT3,Bcl-2 protein,amd decreased the expression of BAX gene.Leptin at different concentrations had a proliferation effect on sheep follicular granulosa cells,and 50 ng/mL had the best effect.Leptin activated the JAK2/STAT3 pathway through interaction with receptors to play an anti-apoptotic role in sheep follicular granulosa cells,which provided a new idea for the treatment of low fertility or infertility caused by metabolic disorders in human and livestock.

关 键 词：绵羊 瘦素 卵泡颗粒细胞 免疫荧光方法 增殖 JAK2/STAT3 

分 类 号：S814.1[农业科学—畜牧学]