畜牧兽医学报2018，Vol.49Issue(4)：701-708,8.DOI:10.11843/j.issn.0366-6964.2018.04.006
TGF-β调节仔猪睾丸支持细胞增殖的机制
Mechanism of TGF-Beta Regulating Proliferation of Boar Sertoli Cells
摘要
Abstract
The objective of the study was to investigate the effect of TGF-β on the expression of proliferation-related proteins and genes of boar Sertoli cells,and reveal the mechanism of TGF-β regulating proliferation of boar Sertoli cells.In this study,TGF-β and TGF-β/Smad pathway inhibitor LY2109761 was used to stimulate cultured immature boar Sertoli cells.Cell viability and cell cycle were detected by CCK-8 kit and flow cytometry,respectively.The phosphorylation levels of Smad3 were detected by Western blotting.The expressions of c-Myc mRNA,Skp2 mRNA and ssc-miRNA-24 were detected by real-time quantitative PCR (RT-qPCR).Furthermore,sscmiRNA-24 mimics and inhibitor were transfected into Sertoli cells,and then Sertoli cells were treated with TGF-β,following detected the expression of related genes and proteins.The results showed that TGF-β influenced cell viability in a dose-dependent manner.180 pg · mL-1 TGF-β increased cell proliferation index(PI)and S-phase cell fraction (SPF),inhibited the phosphorylation of Smad3 in a time-dependent (0-24 h) manner,increased the abundance of c-Myc mRNA,Skp2 mRNA,ssc-miRNA-24.10 μmol · L-1 LY2109761 promoted TGF-β-induced cell viability,increased cell proliferation index and S-phase cell fraction.In addition,ssc-miRNA-24 mimics decreased phosphorylation level of Smad3,promoted the expression of c-Myc and Skp2 mRNA.However,ssc-miRNA-24 inhibitor had an opposite effect on these parameters.The above results revealed that TGF-β decreased phosphorylation level of Smad3 by increasing the expression of sscmiRNA-24,and enhanced the expression of c-Myc and Skp2 mRNA to promote the proliferation of boar Sertoli cells.关键词
TGF-β/支持细胞/增殖/Smad3/ssc-miRNA-24Key words
TGF-β/Sertoli cells/proliferation/Smad3/ssc-miRNA-24分类
农业科技引用本文复制引用
黄莎,孙思,邓洁,胡煜,练雨,王鲜忠..TGF-β调节仔猪睾丸支持细胞增殖的机制[J].畜牧兽医学报,2018,49(4):701-708,8.基金项目
国家自然科学基金项目(31672624 （）
31072183) （）
中央高校基本科研业务费专项资金(XDJK2017D029) （XDJK2017D029）
