白藜芦醇和miR-222协同调控猪卵巢颗粒细胞凋亡  

Resveratrol and miR-222 Synergistically Regulate Apoptosis of Porcine(Sus scrofa)Ovarian Granulosa Cells

作　　者：章会斌 周忍 许黎明 陈一歌 张鹏 杨明月 杨敏 丁月云 殷宗俊 张晓东 ZHANG Hui-Bin;ZHOU Ren;XU Li-Ming;CHEN Yi-Ge;ZHANG Peng;YANG Ming-Yue;YANG Min;DING Yue-Yun;YIN Zong-Jun;ZHANG Xiao-Dong(College of Animal Science and Technology,Anhui Agricultural University/Anhui Provincial Key Laboratory of Genetic Resources Protection and Biological Breeding of Local Livestock and Poultry,Hefei 230036,China)

机构地区：[1]安徽农业大学动物科技学院/地方畜禽遗传资源保护与生物育种安徽省重点实验室,合肥230036

出　　处：《农业生物技术学报》2021年第6期1094-1102,共9页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31972531;31572377;31402037);国家科技支撑计划(2015BAD03B01);农业农村部动物(家禽)遗传育种与繁殖重点实验室开放课题(poultrylab 2019-6)。

摘　　要：卵巢颗粒细胞凋亡影响动物卵泡发育及排卵过程,白藜芦醇在卵巢颗粒细胞凋亡过程中发挥重要作用。本研究旨在解析白藜芦醇介导miR-222调控猪卵巢颗粒细胞(porcine ovarian granulosa cells,pGCs)凋亡的机制,为白藜芦醇在养猪生产中的应用提供理论支持。实验以不同浓度的白藜芦醇(0~100μmol/L)处理分离培养获得的原代pGCs,在倒置显微镜下观察pGCs形态变化,借助流式细胞术检测细胞凋亡率及细胞周期分布,利用qRT-PCR检测细胞周期相关基因CyclinD、CyclinB、p21及miR-222的表达变化;并检测miR-222过表达状态下pGCs凋亡的变化。结果显示当白藜芦醇浓度达到100μmol/L时,p GCs的形态发生改变,细胞增殖率显著下降,细胞周期进程被阻碍;miR-222的表达水平随白藜芦醇浓度的增加呈先下降后上升的趋势,75μmol/L时为拐点(表达量最低);在75μmol/L白藜芦醇处理的pGCs中过表达miR-222,pGCs凋亡率显著下降。以上结果表明白藜芦醇(100μmol/L以下)可抑制pGCs增殖,并以浓度依赖的方式调控细胞中miR-222的表达;且在75μmol/L白藜芦醇处理的pGCs中过表达miR-222,可降低细胞凋亡率。本研究结果为白藜芦醇协同miRNA调控pGCs的生物学功能提供了参考数据。Apoptosis of ovarian granulosa cells affects follicle development and ovulation in animals.Resveratrol plays an important role in the apoptosis of ovarian granulosa cells.This research was aimed to provide data support for the application of resveratrol in pig(Sus scrofa)production by analyzing the mechanism of resveratrol and miR-222 regulate the apoptosis of porcine ovarian granulosa cells(pGCs).The primary pGCs were treated with different concentrations of resveratrol(0~100μmol/L),the morphological changes of pGCs were observed under an inverted microscope,the apoptosis rate and cell cycle distribution were detected by flow cytometry,and the expression of CyclinD,CyclinB,p21 and miR-222 were detected by the qRT-PCR method.The pGCs apoptosis change was also detected in the miR-222 overexpression condition.The results showed that when the concentration of resveratrol reached 100μmol/L,the morphology of pGCs was changed,the apoptosis of pGCs was decreased and the cell cycle of pGCs was inhibited.The expression level of miR-222 was affected by different concentrations of resveratrol and reached the lowest level at 75μmol/L.The pGCs apoptosis was significantly decreased when miR-222 overexpressed in 75μmol/L resveratrol.These results indicated that resveratrol(100μmol/L)could inhibit pGCs proliferation,and regulate miR-222 expression depend on the concentration.Overexpression of miR-222 could decrease pGCs apoptosis with 75μmol/L resveratrol treatment.The results of the study provide more reference data for resveratrol combined miRNA to regulate the biological functions of pGCs.

关 键 词：白藜芦醇 MIR-222 颗粒细胞 猪 

分 类 号：S828[农业科学—畜牧学]