精氨酸对干扰素-tau处理牛子宫内膜上皮细胞基因表达的影响

doi:10.16431/j.cnki.1671-7236.2021.07.025

中国畜牧兽医 ›› 2021, Vol. 48 ›› Issue (7): 2495-2503.doi: 10.16431/j.cnki.1671-7236.2021.07.025

精氨酸对干扰素-tau处理牛子宫内膜上皮细胞基因表达的影响

余婕¹, 胡修忠¹, 向敏¹, 王定发¹, 陶弼菲¹, 夏瑜¹, 陶虎², 熊琪², 程蕾¹

1. 武汉市农业科学院, 武汉 430065;
2. 湖北省农业科学院畜牧兽医研究所, 动物胚胎工程与分子育种湖北省重点实验室, 武汉 430064

收稿日期:2020-11-05 出版日期:2021-07-20 发布日期:2021-07-15
通讯作者: 熊琪, 程蕾 E-mail:phenixxq@163.com;chenglei_011@126.com
作者简介:余婕(1990-),女,湖北十堰人,硕士,畜牧师,研究方向:动物营养与饲料科学,E-mail:yujiehzau@163.com
基金资助:
动物胚胎工程及分子育种湖北省重点实验室开放课题（KLAEMB-2019-05）；武汉市农业科学院创新基金项目（CX201902）

Effects of Arginine on Expression of Genes in Bovine Endometrial Epithelial Cells Treated with Interferon-tau

YU Jie¹, HU Xiuzhong¹, XIANG Min¹, WANG Dingfa¹, TAO Bifei¹, XIA Yu¹, TAO Hu², XIONG Qi², CHENG Lei¹

1. Wuhan Academy of Agricultural Sciences, Wuhan 430065, China;
2. Hubei Key Laboratory of Animal Embryo Engineering and Molecular Breeding, Institute of Animal Science and Veterinary Medicine, Hubei Academy of Agricultural Sciences, Wuhan 430064, China

Received:2020-11-05 Online:2021-07-20 Published:2021-07-15

摘要/Abstract

摘要： 试验旨在研究精氨酸（Arg）对干扰素-tau（interferon-tau，IFNT）处理的牛子宫内膜上皮细胞的调控作用及其可能的机制。将牛子宫内膜上皮细胞接种至6孔细胞培养板中，当达到70%~80%融合度时，向细胞培养基中加入不同浓度的IFNT（0、100 ng/mL），12 h后收集细胞检测未折叠蛋白反应（unfolded protein response，UPR）通路相关基因的表达情况；用不同浓度（0、0.05、0.1、0.2、0.5、1、2 mmol/L）Arg处理牛子宫内膜上皮细胞，24 h后用CCK8法检测其对增殖率的影响，筛选出Arg的最佳作用浓度；将子宫内膜上皮细胞随机分为Arg饥饿组、Arg饥饿+IFNT处理组、Arg添加+IFNT处理组，在Arg饥饿或添加处理6 h后加入100 ng/mL IFNT，IFNT处理12 h后检测UPR通路（BiP、PERK、CHOP、ATF6）、凋亡（BAX、Caspase9）和容受性（HOXA10）相关基因表达情况。结果显示，IFNT刺激可诱导牛子宫内膜上皮细胞UPR通路相关基因BiP、PERK、CHOP、ATF6的表达显著上调；0.5 mmol/L Arg可显著提高牛子宫内膜上皮细胞的增殖率（P<0.05）；Arg缺失可显著上调IFNT诱导下牛子宫内膜上皮细胞中BiP、PERK、CHOP、ATF6和BAX基因mRNA的表达（P<0.05），并显著下调HOXA10基因mRNA的表达（P<0.05），但对Caspase9基因mRNA表达无显著影响（P>0.05）。结果表明，Arg缺失可导致牛子宫内膜上皮细胞发生内质网应激，并影响子宫内膜宫受性的建立，可为进一步揭示Arg对牛子宫内膜的调控机制及制定减少妊娠早期胚胎丢失的营养调控策略提供参考。

关键词: 精氨酸(Arg); 干扰素-tau; 牛子宫内膜上皮细胞; 未折叠蛋白反应

Abstract: This experiment was aimed to investigate the regulatory effect of arginine(Arg) on bovine endometrium treated with interferon-tau(IFNT) and its potential molecular mechanism.Bovine endometrial epithelial cells were inoculated into 6-well cell culture plate,different concentrations of IFNT (0,100 ng/mL) were added into the culture medium after the confluence of cells reached 70% to 80%.Cells were harvested to determine the expression of genes related to unfolded protein response (UPR) pathway after 12 h;Different concentrations of Arg (0,0.05,0.1,0.2,0.5,1,2 mmol/L) were subjected to bovine endometrial epithelial cells to detect the proliferation rate for 24 h by CCK8 method,and the optimal concentration of Arg was selected;Bovine endometrial epithelial cells were randomly divided into Arg starvation group,Arg starvation+IFNT treatment group,Arg+IFNT treatment group,100 ng/mL IFNT was added after Arg deprivation or supplementation for 6 h.The expression of genes associated with UPR pathway (BIP,PERK,CHOP,ATF6),apoptosis (Bax,Caspase9) and endometrial receptivity (HOXA10) were detected after IFNT stimulation for 12 h.The results showed that:IFNT stimulation significantly up-regulated the expression of BiP,PERK,CHOP and ATF6 genes implicated in UPR pathway in bovine endometrial epithelial cells.The proliferation rate of bovine endometrial epithelial cells were significantly increased by 0.5 mmol/L Arg (P<0.05);Arg deprivation significantly up-regulated the mRNA level of Bip,PERK,CHOP,ATF6 and BAX genes in IFNT-treated bovine endometrial epithelial cells (P<0.05),and significantly down-regulated the mRNA level of HOXA10 (P<0.05),but had no significant effect on the expression of Caspase9 mRNA (P>0.05).The results indicated that Arg deficiency could lead to endoplasmic reticulum stress in bovine endometrial epithelial cells and influence the establishment of endometrial receptivity,which could provide reference for further revealing the regulation mechanism of Arg on bovine endometrium and formulating nutritional regulation strategies to reduce embryo loss in early pregnancy.

Key words: arginine (Arg); interferon-tau; bovine endometrial epithelial cell; unfolded protein response

中图分类号:

S814.8

余婕, 胡修忠, 向敏, 王定发, 陶弼菲, 夏瑜, 陶虎, 熊琪, 程蕾. 精氨酸对干扰素-tau处理牛子宫内膜上皮细胞基因表达的影响[J]. 中国畜牧兽医, 2021, 48(7): 2495-2503.

YU Jie, HU Xiuzhong, XIANG Min, WANG Dingfa, TAO Bifei, XIA Yu, TAO Hu, XIONG Qi, CHENG Lei. Effects of Arginine on Expression of Genes in Bovine Endometrial Epithelial Cells Treated with Interferon-tau[J]. China Animal Husbandry and Veterinary Medicine, 2021, 48(7): 2495-2503.

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精氨酸对干扰素-tau处理牛子宫内膜上皮细胞基因表达的影响

Effects of Arginine on Expression of Genes in Bovine Endometrial Epithelial Cells Treated with Interferon-tau

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