摘要
目的:探讨不同褪黑素浓度下处理不同时间对牛颗粒细胞凋亡相关基因表达的影响.方法:牛颗粒细胞从卵巢卵泡(直径3～8 mm)中分离并培养,培养3代后用于试验,分别在培养液中加入终浓度为0、10-3、10-5、10-7、10-9、10-11 mol/L褪黑素,培养24、48、72 h后,分别收集细胞,提取总RNA,利用实时定量PCR方法检测各组细胞中抗凋亡(BCL2和BCL-XL)、促凋亡(BAX、TP53和CASP3)基因表达情况.结果:不同剂量褪黑素作用24、48、72 h,可显著提高牛颗粒细胞中BCL2和BCL-XL的表达水平,抑制BAX、TP53、CASP3基因表达(P＜0.05),但10-3、10-5 mol/L组的BCL-XL表达量均无显著性变化(P＞0.05),而72 h时,10-7、10-9、10-11 mol/L组TP53基因表达显著增加(P＜0.05).结论:不同浓度褪黑素处理不同时间对牛颗粒细胞中凋亡相关基因表达存在不同影响,褪黑素促进抗凋亡基因和抑制促凋亡基因的表达.
Abstract
Objective:To investigate the effects of different melatonin concentrations at different times on the expression of apoptosis-related genes in bovine granulosa cells.Methods:Bovine granulosa cells were isolated and cultured from bovine ovarian follicles(3-8 mm in diameter).Bovine granulosa cells were cultured for three generations and then entered the experiment.Melatonin of 0,10-3,10-5,10-7,10-9,10-11 mol/L was added to the culture medium.After 24,48,72 h of culture,the cells were collected and total RNA was extracted.The expressions of anti-apoptotic(BCL2 and BCL-XL)and pro-apoptotic(BAX,TP53 and CASP3)genes in each group were detected by real-time PCR.Results:The effects of different doses of melatonin for 24,48,72 h significantly promoted the expression levels of BCL2 and BCL-XL and inhibited the expression of BAX,TP53 and CASP3 genes in bovine granulosa cells(P＜0.05),but there was no significant change in the expression of BCL-XL in both the 10-3,10-5 mol/L groups(P＞0.05),whereas at 72 h,the 10-7,10-9,10-11 mol/L groups showed a significant increase in TP53 gene expression at 72 h(P＜0.05).Conclusion:Different concentrations and time of melatonin treatment have different effects on apoptosis-related genes in bovine granulosa cells.Melatonin promotes the expression of anti-apoptotic genes and inhibits the expression of pro-apoptotic genes.
基金项目
安徽省自然科学基金(2008085MC94)
安徽省重点研发计划(202204c06020076)
安徽省高等学校自然科学研究项目(KJ2021A0869)
维普
万方数据