中国农业科学2025，Vol.58Issue(7)：1451-1462,12.DOI:10.3864/j.issn.0578-1752.2025.07.015
谷氨酰胺对氧化应激状态下胎衣不下奶牛内质网应激的影响
Effect of Gln on Endoplasmic Reticulum Stress in Retained Fetal Membranes Cows Under Oxidative Stress via the PI3K/AKT Pathway
摘要
Abstract
[Objective]This study aimed to determine whether glutamine could alleviate endoplasmic reticulum stress in the maternal placenta of cows with retained fetal membranes(RFM)caused by oxidative stress through the PI3K pathway,and to explore the protective mechanism of glutamine.[Method]Three cows with RFM and three healthy cows(NRFM)were selected based on whether the fetal membranes could be normally expelled within 12 hours after parturition.The content of glutamine(Gln)in the serum of RFM and NRFM cows was determined.The expression changes of endoplasmic reticulum stress(GRP78,p-PERK,PERK,p-IRE1α,IRE1α,and ATF6)and key apoptosis factors(Caspase-3,Caspase-8,Caspase-9,P53,Bcl-2,and Bax)in the maternal placenta of the two groups of cows were detected by qRT-PCR and Western blot.In the in vitro experiment,bovine uterine caruncle epithelial cells(UCEs)were stimulated with 400 μmol·L-1 H2O2 to establish an in vitro oxidative stress model of bovine UCEs.On this basis,Gln pretreatment,PI3K inhibitor(LY294002)and glutamine co-pretreatment were performed.The concentration changes of Ca2+in the cytoplasm were detected by immunofluorescence,and the expression changes of endoplasmic reticulum stress and apoptosis-related indicators in bovine UCEs were detected by qRT-PCR and Western blot.[Result]When cows had RFM,the serum Gln content was significantly decreased(P＜0.01),and the expression of endoplasmic reticulum stress-related proteins and genes GRP78(P＜0.01),p-PERK/PERK,p-IRE1α/IRE1α,ATF6(P＜0.05)in the placenta tissue was significantly increased,while the expression of apoptosis-related proteins and genes P53(P＜0.01),Caspase-3,Caspase-8,Caspase-9,Bax/Bcl-2(P＜0.05)was significantly decreased.In the in vitro oxidative stress model,the expression of endoplasmic reticulum stress-related proteins and genes GRP78,p-PERK/PERK,p-IRE1α/IRE1α,ATF6(P＜0.01)and apoptosis-related proteins and genes P53,Caspase-3,Caspase-8,Caspase-9,Bax/Bcl-2(P＜0.01)in UCEs were significantly increased after H2O2 stimulation,and the concentration of Ca2+in the cytoplasm was significantly increased(P＜0.01).However,compared with the Gln protection group pretreated only with glutamine,Gln could not reduce the expression of endoplasmic reticulum stress and apoptosis-related proteins and genes in bovine UCEs when PI3K was inhibited,and the concentration of Ca2+in the cytoplasm was significantly increased(P＜0.01).[Conclusion]When cows had RFM,the maternal placenta tissue underwent strong endoplasmic reticulum stress and abnormal apoptosis,which led to the damage of normal cell function and was an important cause of the disorder of fetal membrane expulsion.High levels of Gln could regulate the expression of key proteins in endoplasmic reticulum stress through the PI3K/AKT pathway,alleviate the cellular dysfunction caused by endoplasmic reticulum stress,and thereby reduce the placental expulsion disorder caused by endoplasmic reticulum stress induced by oxidative stress.关键词
胎衣不下/氧化应激/内质网应激/谷氨酰胺/奶牛子宫内膜上皮细胞Key words
retained fetal membranes/oxidative stress/endoplasmic reticulum stress/glutamine/uterine caruncle epithelial引用本文复制引用
王薇,罗春海,贾红豆,刘佳金,李丹阳,付世新..谷氨酰胺对氧化应激状态下胎衣不下奶牛内质网应激的影响[J].中国农业科学,2025,58(7):1451-1462,12.基金项目
国家自然科学基金(32473102)、国家"十四五"重点研发计划(2023YFD1801100)、黑龙江八一农垦大学研究生创新科研项目(YJSCX2023-Y34) （32473102）
