miR-96对牦牛颗粒细胞凋亡和增殖的影响
Effects of miR-96 on the apoptosis and proliferation of Bos grunniens granulosa cells
贾跃宁 1彭巍 2徐尚荣 2刘海婷 1黄荣 2舒适 2王国文 2付长其 2张君2
作者信息
- 1. 青海大学农牧学院,青海西宁 810016
- 2. 青海大学畜牧兽医科学院,青海西宁 810016
- 折叠
摘要
为研究牦牛卵巢组织内miR-96对颗粒细胞凋亡、增殖及激素分泌的影响,本试验通过采集成年牦牛卵巢内颗粒细胞,分离培养后将其分为4个处理组,分别转染miR-96模拟物(mimics)和抑制物(inhibitor)及两者的阴性对照(NC)后培养并检测颗粒细胞受到的影响.使用流式细胞术和CCK-8法对细胞凋亡和增殖情况进行检测,采用qPCR和Western Blot检测凋亡相关基因表达情况,ELISA检测转染后类固醇激素分泌情况.结果表明:miR-96-mim-ics使颗粒细胞凋亡发生抑制促进细胞增殖,并促使凋亡相关基因Bcl-2表达极显著升高,Bax表达极显著降低;ELISA检测类固醇激素水平变化显示,与mimics NC组相比miR-96-mimics组细胞培养液中孕酮(P4)分泌水平极显著降低,雌二醇(E2)分泌水平极显著升高;与inhibitor NC组相比miR-96-inhibitor组孕酮(P4)分泌水平极显著升高,雌二醇(E2)分泌水平显著降低.miR-96可促进颗粒细胞增殖并抑制凋亡,同时会对生殖激素分泌产生影响,从而参与调控牦牛卵泡发育与闭锁的过程.
Abstract
The granulosa cells collected from adult Bos grunniens ovaries are taken as research mate-rials to investigate the effects of miR-96 on the apoptosis,proliferation and hormone secretion of the cells in Bos grunniens ovarian tissue.The cells are divided into four groups after isolation and cultiva-tion,transfected with miR-96 mimics and inhibitors as well as their negative controls(NC).The effects on the cells are cultured and detected as well.Flow cytometry and CCK-8 assay are used to detect cell apoptosis and proliferation.Besides,qPCR and Western Blot are used to detect the expres-sion of apoptosis-related genes.The steroid hormone secretion after transfection is detected by using ELISA.The results show that miR-96-mimics group inhibit granulosa cells apoptosis and promote cell proliferation.In addition,it significantly increases the expression of apoptosis-related gene and significantly decreases the expression of Bax.According to the variations steroid hormone levels de-tected by ELISA,it can be concluded that compared with mimics NC group,the secretion level of P4 in the cell culture medium of miR-96-mimics group is significantly reduced,while the secretion level of E2 is significantly increased.Compared with inhibitor group,the secretion level of P4 of miR-96-inhibitor group is significantly rised,while that of E2 is significantly decreased.In summa-ry,miR-96 can promote granulosa cell proliferation,inhibit apoptosis and affect reproductive hor-mone secretion,thereby participating in the regulation of follicular development and occlusion of Bos grunniens.
关键词
牦牛/miR-96/颗粒细胞/凋亡/增殖Key words
Bos grunniens/miR-96/granulosa cell/apoptosis/proliferation引用本文复制引用
基金项目
青海省科技厅项目(2022-NK-119)
青海省昆仑英才高端创新创业人才计划培养领军人才项目(2021)()
出版年
2024
NETL
NSTL
万方数据