摘要: |
旨在研究FSH-AKT-FOXO1途径调控绵羊卵泡颗粒细胞增殖机制。利用不同浓度FSH处理绵羊颗粒细胞(GCs),采用CCK-8法检测细胞活性,流式细胞仪测定细胞凋亡和细胞周期变化,Western-Blot检测p-AKT和p-FOXO1蛋白质表达,qPCR分析PCNA、CCnd-2和Bcl-2基因表达量变化。结果表明:10 ng/mL 的FSH能显著提高GCs活性,抑制细胞凋亡(P<0.05);AKT/FOXO信号通路的抑制剂AT7867预处理GCs后明显降低其活性(P<0.05);在FSH的作用下,GCs中AKT蛋白的磷酸化水平显著升高,而FOXO1的磷酸化水平显著降低(P<0.05),同时显著上调了PCNA、CCnd-2和Bcl-2 的表达水平(P<0.05)。本研究表明FSH通过 AKT/FOXO1 信号途径促进了绵羊GCs的生长、增殖。 |
关键词: 绵羊 颗粒细胞 FSH AKT/FOXO1信号途径 绵羊 |
DOI:10.11841/j.issn.1007-4333.2020.03.06 |
投稿时间:2019-09-28 |
基金项目:国家自然科学基金项目(31460598) |
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Follicle-stimulating hormone regulating the proliferation of ovine follicular granulosa cells through AKT/FOXO1 pathway |
LIU Chunjie,WANG Zhaochen,DU Wei,ZHAO Yongchao,ZHANG Jiaxin
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(College of Animal Science/Key Laboratory of Animal Genetics、Breeding and Reproduction of Inner Mongolia Autonomous Region, Inner Mongolia Agricultural University, Hohhot 010018, China) |
Abstract: |
The purpose of this study was to investigate the proliferation of ovine follicular granulosa cells(GCs)regulated by FSH-AKT-FOXO1 pathway. After GCs were treated with difference concentrations FSH, the cells' viability were measured by CCK-8assays. The apoptosis and cell cyclewere were assayed by Flowcytometry. The phosphorylation level of AKT and FOXO1 protein were detected by Western-Blot. The transcriptional profile of PCNA, CCnd-2 and Bcl-2 were detected by qRT-PCR. The results showed that: adding 10 ng/mL FSH in the culturemedium significantly increased the viability of cells and inhibited apoptosis(P<0. 05). The viability of GCs decreased significantly after pre-treatment with the inhibitor AT7867 of AKT/FOXO1 signal pathway(P<0. 05). FSH significantly promoted AKT phosphorylation and decreased FOXO1 phosphorylation, FSH significantly up-regulated the expressions of CCnd-2, PCNA and Bcl-2 mRNA. In conclusion, FSH promoted the growth and proliferation of ovine follicular granulosa cells through AKT/FOXO1 pathway. |
Key words: ovine granulosa cell FSH AKT/FOXO1 signal pathway |