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

Abstract

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

CLC Number:

S854.43

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.

Figures/Tables 6

Table 1

Fig. 1

Fig. 2

Fig. 3

Fig. 4

Fig. 5

References 23

1	KELTOND F,LISSEMOREK D,MARTINR E.Recommendations for recording and calculating the incidence of selected clinical diseases of dairy cattle[J].J Dairy Sci,1998,81(9):2502-2509. doi: 10.3168/jds.S0022-0302(98)70142-0
2	刘羽. 规模牧场奶牛围产后期主要疾病病因调查分析[D]. 泰安: 山东农业大学, 2023.
	LIU Y. Investigation and analysis of the main causes of diseases in dairy cows during the late stage of perinatal in large-scale ranches[D]. Taian: Shandong Agricultural University, 2023. (in Chinese)
3	汤兆鹏,王冠男,周子凡,等.奶牛胎衣不下的诊断和治疗[J].畜牧兽医杂志,2024,43(3):127-130.
	TANGZ P,WANGG N,ZHOUZ F,et al.Diagnosis and treatment of retained placenta indairy cows[J].Journal of Animal Science and Veterinary Medicine,2024,43(3):127-130.
4	宋玉锡. 亚临床酮病奶牛产后乏情的关键蛋白筛选及作用机制研究[D]. 大庆: 黑龙江八一农垦大学, 2024.
	SONG Y X. Screening of key proteins and study on its action mechanism of dairy cows with postpartum anestrus caused by subclinical ketosis[D]. Daqing: Heilongjiang Bayi Agricultural University, 2024. (in Chinese)
5	邢菲菲. 高NEFA对奶牛子宫内膜上皮细胞ECM降解关键因子的影响[D]. 大庆: 黑龙江八一农垦大学, 2023.
	XING F F. Effect of high concentration NEFA on key factors of ECM degradation in endometrial epithelial cells of dairy cows[D]. Daqing: Heilongjiang Bayi Agricultural University, 2023. (in Chinese)
6	ROUPEK M,VEERLAS,OLSONJ,et al.Transcription factor binding site analysis identifies FOXO transcription factors as regulators of the cutaneous wound healing process[J].PLoS One,2014,9(2):e89274. doi: 10.1371/journal.pone.0089274
7	LINKW.Introduction to FOXO biology[J].Methods Mol Biol,2019,1890,1-9.
8	HENRIQUESV,MACHADOS,LINKW,et al.Monitoring the transcriptional activity of FOXO transcription factors by analyzing their target genes[J].Methods Mol Biol,2019,1890,103-113.
9	MEIZ G,HUANGY G,FENGZ T,et al.Electroacupuncture ameliorates cerebral ischemia/reperfusion injury by suppressing autophagy via the SIRT1-FOXO1 signaling pathway[J].Aging,2020,12(13):13187-13205. doi: 10.18632/aging.103420
10	MOTTAM C,DIVECHAN,LEMIEUXM,et al.Mammalian SIRT1 represses forkhead transcription factors[J].Cell,2004,116(4):551-563. doi: 10.1016/S0092-8674(04)00126-6
11	温小庆. 高NEFA经FoxO1途径诱导奶牛子宫内膜上皮细胞凋亡的调节机制[D]. 大庆: 黑龙江八一农垦大学, 2024.
	WEN X Q. Mechanism regulation of bovine endometrial epithelial cells apoptosis induced by high NEFA via FoxO1 pathway[D]. Daqing: Heilongjiang Bayi Agricultural University, 2024. (in Chinese)
12	郑程远. miRNA-185调控VEGFA信号通路影响奶牛胎衣不下的分子机制[D]. 大庆: 黑龙江八一农垦大学, 2018.
	ZHENG C Y. miRNA-185 regulates the VEGFA signaling pathway in dairy cows with retained fetal membranes[D]. Daqing: Heilongjiang Bayi Agricultural University, 2018. (in Chinese)
13	PAUDYALS,MAUNSELLF,SMITHJ,et al.Association of rumination time with non-esterified fatty acid, β-hydroxybutyrate, and serum calcium concentrations in transition dairy cows[J].AABP Conference Proc,2015,210-211.
14	曹丽光,刘思宇,栾奕,等.白藜芦醇缓解NEFA诱导的奶牛乳腺上皮细胞的周期阻滞和凋亡[J].中国兽医学报,2024,44(5):1032-1040.
	CAOL G,LIUS Y,LUANY,et al.Resveratrolalleviates NEFA-induced cellcyclearrestand apoptosisin bovine mammary epithelial cells[J].Chinese Journal of Veterinary Science,2024,44(5):1032-1040.
15	哈依努尔,李俊,陈永恒,等.翼螺旋转录因子FOXO1的DNA结合域的表达、纯化及结合特性[J].中南大学学报,2017,42(1):1-7.
	HAY N E,LIJ,CHENY H,et al.Expression and purification of FOXO1 DNA binding domain and its DNA properties[J].Journal of Central South University(Science and Technology),2017,42(1):1-7.
16	RODRIGUEZ-COLMANM J,DANSENT B,BURGERINGM T.FOXO transcription factors as mediators of stress adaptation[J].Nat Rev Mol Cell Biol,2024,25(1):46-64. doi: 10.1038/s41580-023-00649-0
17	MEGIDA I A,KAMAL EL-DINM M M,MORSHEDYN A,et al.The association between silent mating type information regulator 2 homolog 1 (SIRT1) gene polymorphism and systemic lupus erythromatosis (SLE)[J].QJM: Int J Med,2021,114(Supplement_1):hcab091.013. doi: 10.1093/qjmed/hcab091.013
18	ZHANGM,ZHANGQ,HUY,et al.miR-181a increases FoxO1 acetylation and promotes granulosa cell apoptosis via SIRT1 downregulation[J].Cell Death Dis,2017,8(10):e3088-e3088. doi: 10.1038/cddis.2017.467
19	张洪开,徐韬钧,具英花,等.PI3K和MAPK信号通路通过FOXO1转录因子诱导A549细胞周期的阻滞和凋亡的研究[J].中国医科大学学报,2012,41(10):908-911.
	ZHANGH K,XUT J,JUY H,et al.PI3K/AKT and MAPK/ERK pathways induce cell cycle arrest and apoptosis in A549 cell through the regulation of FOXO1 transcription factor[J].Journal of China Medical University,2012,41(10):908-911.
20	车敏,张辉.紫铆因通过激活SIRT1介导FOXO1/NF-κB信号通路改善坐骨神经损伤[J].中国医科大学学报,2024,53(2):102-107.
	CHEM,ZHANGH.Butein ameliorates sciatic nerve injury by activating SIRT1 and mediating the FOXO1/NF-κB signaling pathway[J].Journal of China Medical University,2024,53(2):102-107.
21	张国华. 系统性红斑狼疮外周血CD4⁺ T细胞Sirt1的表达及Resveratrol对CD4⁺ T细胞亚群及功能影响的初探[D]. 北京: 北京协和医学院, 2011.
	ZHANG G H. Expression of Sirt1 in CD4⁺ T cell and the effect of Resveratrol on percentage of CD4⁺ T cell subgroups and their cytokines and on fuction of CD4⁺ T cell and PBMC isolated from peripheral blood of patients with systemic lupus erythematosus[D]. Beijing: Peking Union Medical College, 2011. (in Chinese)
22	刘远波,王霞,包太成,等.白藜芦醇激活SIRT1抑制高糖诱导的神经细胞凋亡[J].中南医学科学杂志,2021,49(2):152-157.
	LIUY B,WANGX,BAOT C,et al.Resveratrol inhibits HG-induced apoptosis of PC12 cell by SIRT1 activation[J].Medical Science Journal of Central South China,2021,49(2):152-157.
23	LINK N,ZHANGK,ZHAOW,et al.Insulin-like growth factor 1 promotes cell proliferation by downregulation of G-protein-coupled receptor 17 expression via PI3K/Akt/FoxO1 signaling in SK-N-SH Cells[J].Int J Mol Sci,2022,23(3):1513. doi: 10.3390/ijms23031513

基因名称 Gene	引物序列(5′→ 3′) Primer sequences	片段大小/bp Fragment length
SIRT1	F：GAAGACAATGGGCCGGGCTTAC R：GCCGCCTCTTCCTCCTCCTC	3 956
β-actin	F：TCCTGCGGCATTCACGAAACTAC R：GTGTTGGCGTAGAGGTCCTTGC	1 948

The Influence of FoxO1 on Expression of Apoptotic Factors in Dairy Cow Endometrial Epithelial Cells Induced by High NEFA Levels

RichHTML

PDF (PC)

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 6

References 23

Related Articles 1

Recommended Articles

Metrics

Comments