中国畜牧兽医2024，Vol.51Issue(5)：2122-2131,10.DOI:10.16431/j.cnki.1671-7236.2024.05.033
表没食子儿茶素没食子酸酯对脂多糖诱导的奶牛乳腺上皮细胞炎性损伤和凋亡的干预作用
Effects of EGCG on Inflammatory Injury and Apoptosis of BMECs Induced by LPS
摘要
Abstract
[Objective]The aim of this experiment was to investigate the effects of epigallocatechin gallate(EGCG)on lipopolysaccharide(LPS)induced inflammation and apoptosis in bovine mammary epithelial cells(BMECs)and its potential protective mechanisms.[Method]The cell inflammation model was constructed by LPS-induced BMECs.The cell viability was detected by CCK-8 and the optimal concentration of EGCG was screened.BMECs were divided into control group,LPS treatment group and EGCG+LPS co-treatment group.Real-time quantitative PCR was used to detect the mRNA expression levels of inflammatory factors,apoptosis factors and anti-oxidation-related genes.Mitochondrial membrane potential and reactive oxygen species(ROS)levels of BMECs were detected by kit.The apoptosis of BMECs was detected by flow cytometry.[Result]After LPS-induced BMECs,the mRNA expressions of inflammatory factors interleukin-6(IL-6),IL-8 and IL-1β genes were extremely significantly or significantly increased at all time periods compared with 0 h(P＜0.01 or P＜0.05),the cell inflammatory damage model of BMECs was successfully constructed.CCK-8 assay showed that the optimal concentration of EGCG was 5 μmol/L.Real-time quantitative PCR results showed that,compared with control group,the mRNA expressions of IL-6,IL-8,MDA,Bax and Caspase-3 genes in LPS-induced BMECs were extremely significantly increased(P＜0.01),while the mRNA expressions of GSH-Px and Bcl-2 genes were extremely significantly decreased(P＜0.01).Compared with LPS group,mRNA expressions of IL-6,IL-8,IL-1β,MDA,Bax and Caspase-3 genes were significantly or extremely significantly decreased in EGCG+LPS group(P＜0.05 or P＜0.01),and the mRNA expressions of GSH-Px,SOD and Bcl-2 gene were significantly or extremely significantly increased(P＜0.05 or P＜0.01).The results of mitochondrial membrane potential showed that compared with control group,LPS significantly weakened the red fluorescence and enhanced the green fluorescence of BMECs,while the co-incubation of EGCG and LPS had no significant effect on the red fluorescence of BMECs,and the green fluorescence was relatively weakened.ROS level detection results showed that compared with control group,the green fluorescence of cells in LPS group was significantly enhanced,while the green fluorescence of cells was weakened after co-incubation of EGCG and LPS.Flow cytometry showed that compared with control group,death cells and apoptotic cells increased significantly after LPS treatment,the apoptosis rate was extremely significantly increased(P＜0.01).After co-incubation with EGCG and LPS,the apoptotic cells were significantly decreased,and the apoptotic rate was significantly decreased(P＜0.05).[Conclusion]EGCG could alleviate inflammatory response and inhibit cell apoptosis by inhibiting ROS release induced by LPS in BMECs and alleviating mitochondrial damage.The results provide a theoretical basis for the prevention and treatment of mastitis in dairy cows with EGCG.关键词
奶牛乳腺上皮细胞(BMECs)/表没食子儿茶素没食子酸酯(EGCG)/脂多糖(LPS)/炎性损伤/细胞凋亡Key words
bovine mammary epithelial cells(BMECs)/epigallocatechin gallate(EGCG)/lipopolysaccharide(LPS)/inflammatory injury/apoptosis分类
农业科技引用本文复制引用
宝华,户春丽,安彦昊,张春华,马燕芬..表没食子儿茶素没食子酸酯对脂多糖诱导的奶牛乳腺上皮细胞炎性损伤和凋亡的干预作用[J].中国畜牧兽医,2024,51(5):2122-2131,10.基金项目
内蒙古自然科学基金项目(2021MS03068) （2021MS03068）
宁夏自然科学基金项目(2022AAC02006) （2022AAC02006）
