摘要
为揭示高NEFA经FoxO1途径诱导奶牛子宫内膜上皮细胞凋亡的机制,研究通过添加0、0.2、1.2 mM的NEFA刺激奶牛子宫内膜上皮细胞12 h,应用流式细胞技术检测细胞凋亡和细胞周期情况,应用WB和qRT-PCR技术检测FoxO1、pFoxO1、Caspase-3、Bcl-2和Bax的蛋白和mRNA表达水平。结果显示:NEFA在低浓度0.2 m M时细胞凋亡无明显变化,在高浓度1.2 mM时细胞凋亡极显著增多(P<0.01);NEFA在高浓度1.2 mM时极显著上调G1期细胞数量,显著下调G2期细胞数量;NEFA在低浓度0.2 mM时对细胞内Caspase-3、Bcl-2、Bax、FoxO1和p-FoxO1蛋白表达无显著影响(P>0.05);NEFA在1.2 mM时极显著上调细胞内Caspase-3、Bax、FoxO1和p-FoxO1的蛋白表达(P<0.01),NEFA在高浓度1.2 mM时极显著下调Bcl-2的蛋白表达(P<0.01)。NEFA在低浓度0.2 mM时对细胞内Caspase-3、Bcl-2、Bax、FoxO1的mRNA表达无显著影响(P>0.05);NEFA在1.2 mM时极显著上调细胞内Caspase-3、Bax和FoxO1的mRNA表达水平(P<0.01),极显著下调Bcl-2的mRNA水平(P<0.01)。研究表明,高浓度NEFA可以通过FoxO1途径诱导奶牛子宫内膜上皮细胞凋亡和细胞周期阻滞。
In order to reveal the mechanism of apoptosis induced by high NEFA through FoxO1 pathway,the endometrial epithelial cells of dairy cows were stimulated by adding 0 mM,0.2 mM and 1.2 mM NEFA for 12 h,and apoptosis and cell cycle were detected by flow cytometry.The protein and mRNA expression levels of FoxO1,p-FoxO1,Caspase-3,Bcl-2 and Bax were detected by WB and qRT-PCR.The results showed that apoptosis of NEFA cells at low concentration of 0.2 mM did not change significantly,and apoptosis of NEFA cells at high concentration of 1.2 mM significantly increased(P<0.01).At a high concentration of 1.2 mM,NEFA significantly increased the number of cells in G1 phase and significantly decreased the number of cells in G2 phase.NEFA at low concentration of 0.2 mM had no significant effect on the expression of Caspase-3,Bcl-2,Bax,FoxO1 and P-FoxO1 proteins in cells(P<0.05);NEFA significantly up-regulated the expression of Caspase-3,Bax,FoxO1 and P-FoxO1 in cells at 1.2 mM(P<0.01),NEFA significantly down-regulated the expression of Bcl-2 protein at a high concentration of 1.2 mM(P<0.01).NEFA at low concentration of 0.2 mM had no significant effect on mRNA expression of Caspase-3,Bcl-2,Bax and FoxO1 in cells(P<0.05);NEFA significantly up-regulated mRNA expression levels of Caspase-3,Bax and FoxO1 in cells at 1.2 mM(P<0.01),significantly down-regulated the mRNA level of Bcl-2(P<0.01).Studies showed that high concentrations of NEFA could induce apoptosis and cell cycle arrest of cow endometrial epithelial cells through FoxO1 pathway.
作者
温小庆
罗春海
曹一鸣
李美娟
付世新
Wen Xiaoqing;Luo Chunhai;Cao Yiming;Li Meijuan;Fu Shixin(College of Animal Science and Technology,Heilongjiang Bayi Agricultural University,Daqing163319)
出处
《黑龙江八一农垦大学学报》
2024年第5期56-62,共7页
journal of heilongjiang bayi agricultural university
基金
黑龙江省自然科学基金(LH2021C073)。
