FoxO1对高NEFA诱导的奶牛子宫内膜上皮细胞凋亡因子表达的影响

doi:10.11843/j.issn.0366-6964.2025.09.047

摘要/Abstract

摘要： 旨在探究叉头盒蛋白O1(fork head box O 1, FoxO1)对高浓度非酯化脂肪酸(non-esterified fatty acids, NEFA)状态下的奶牛子宫内膜上皮细胞凋亡因子表达变化的影响。通过实时荧光定量PCR法及Western blot法评价添加激活剂白藜芦醇(resveratrol)后沉默信息调节因子(silent information regulator 1, SIRT1)的表达效果，明确奶牛子宫内膜上皮细胞中FoxO1的表达情况。加入Resveratrol和高浓度的NEFA后，检测FoxO1通路关键因子及细胞凋亡相关因子蛋白的表达变化情况。与对照组相比，添加Resveratrol上调SIRT1的蛋白和mRNA表达水平(P<0.01)，下调Ac-FoxO1蛋白表达水平(P<0.01)；下调Caspase-3蛋白表达水平无显著影响(P>0.05)，上调Bcl-2、Bcl-xl的蛋白表达水平(P<0.01)，下调Bax和Bax/Bcl-2的蛋白表达水平(P<0.05)。与NEFA组相比，NEFA+Resveratrol组上调IGF-1、SIRT1和p-AKT蛋白表达水平(P<0.01)，下调FoxO1、p-FoxO1、Ac-FoxO1、FoxO1/p-FoxO1、AKT和AKT/p-AKT蛋白表达水平(P<0.01)，上调Bcl-xl和Bcl-2蛋白表达水平(P<0.01)，下调Caspase-3、Bax和Bax/Bcl-2蛋白表达水平(P<0.01)。激光共聚焦检测结果显示，NEFA+Resveratrol组与NEFA组相比，抑制FoxO1的表达水平(P<0.01)。Resveratrol可有效激活SIRT1的表达，进而抑制FoxO1的表达，降低高浓度NEFA状态下奶牛子宫内膜上皮细胞中凋亡因子的表达水平。

关键词: 奶牛子宫内膜上皮细胞, 非酯化脂肪酸, 沉默信息调节因子1, 叉头盒蛋白O1, 凋亡因子

Abstract: Changes in the expression of apoptotic factors in endometrial epithelial cells of dairy cows in the state of high concentration of non-esterified fatty acids (NEFA) were explored to be influenced by forkhead box protein O1 (FoxO1). The effect of silent information regulator 1 (SIRT1) expression was evaluated by real-time fluorescence quantitative PCR and Western blot after the addition of the activator resveratrol (Resveratrol), the expression profile of FoxO1 was clarified in the endometrial epithelial cells of dairy cows. Changes in the expression of key factors of the FoxO1 pathway and apoptosis-associated factor proteins were examined after the addition of Resveratrol and high concentrations of NEFA. The protein and mRNA expression levels of SIRT1 were up-regulated (P<0.01) and Ac-FoxO1 protein expression levels were down-regulated (P<0.01) by the addition of Resveratrol compared with the control group; Caspase-3 protein expression levels were not significantly affected, showing a down-regulation trend but not statistically different (P>0.05); the protein expression levels of Bcl-2 and Bcl-xl were up-regulated (P<0.01); the protein expression levels of Bax and Bax/Bcl-2 were down-regulated (P<0.05). Compared with the NEFA group, in the NEFA+Resveratrol group, IGF-1, SIRT1 and p-AKT protein expression levels were up-regulated (P<0.01); FoxO1, p-FoxO1, Ac-FoxO1, FoxO1/p-FoxO1, AKT and AKT/p-AKT protein expression levels were down-regulated (P<0.01). Bcl-xl and Bcl-2 protein expression levels were upregulated (P<0.01), Caspase-3, Bax and Bax/Bcl-2 protein expression levels were downregulated (P<0.01). The results of laser confocal assay showed that the expression level of FoxO1 was suppressed in the NEFA+Resveratrol group compared with the NEFA group (P<0.01). SIRT1 expression can be effectively activated by Resveratrol, FoxO1 expression was suppressed, and the expression level of apoptotic factors in the endometrial epithelial cells of dairy cows was reduced in the state of high concentration of NEFA.

Key words: bovine endometrial epithelial cells, non-esterified fatty acids, silent information regulator 1, forkhead box protein O1, apoptosis factors

中图分类号:

S854.43

刘佳金, 温小庆, 罗春海, 贾红豆, 王薇, 李丹阳, 付世新. FoxO1对高NEFA诱导的奶牛子宫内膜上皮细胞凋亡因子表达的影响[J]. 畜牧兽医学报, 2025, 56(9): 4708-4717.

LIU Jiajin, WEN Xiaoqing, LUO Chunhai, JIA Hongdou, WANG Wei, LI Danyang, FU Shixin. The Influence of FoxO1 on Expression of Apoptotic Factors in Dairy Cow Endometrial Epithelial Cells Induced by High NEFA Levels[J]. Acta Veterinaria et Zootechnica Sinica, 2025, 56(9): 4708-4717.

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