[Objective]The aim of this study was to investigate the inhibitory effects of hesperidin on apoptosis of bo-vine mammary epithelial cells and its molecular mechanism,so as to provide a theoretical basis for its application in pro-moting the growth and development of mammary gland and maintaining lactation function in cows.[Method]Apoptosis model of bovine mammary epithelial cells was generated by treating it with 1000 μmol/L H2O2 for 8 h,and then treated with 120 μg/mL hesperidin for 24 h.Cell viability was measured by CCK-8 assay,and apoptosis was detected by confocal microscopy,flow cytometer and DNA gel electrophoresis.The expression levels of Bcl-2,Bax and Caspase3 genes were measured by real-time fluorescence quantitative PCR and the protein expression levels of Bcl-2,Bax and Caspase3 were measured by Western blotting.[Result]Compared with the blank control group(CK),the bovine mammary epithelial cell viability was decreased extremely significantly(P<0.01,the same below)and the apoptosis proportion was extremely sig-nificantly increased in H2O2 group,with the DNA fragments appearing as a ladder pattern;the bovine mammary epithelial cell viability was improved significantly in hesperidin group(P<0.05,the same below),while apoptosis rate was de-creased and the number of apoptotic cells was lower in hesperidin group.Compared with H2O2 group,the bovine mam-mary epithelial cell viability was significantly increased and apoptosis rate was extremely significantly decreased,and DNA ladder fragments were decreased in H2O2+hesperidin group.In terms of genes expression of Bax,Bcl-2,and Cas-pase3 and proteins expression,in H2O2 group,Bax and Caspase3 genes expression and protein expression levels were sig-nificantly up-regulated,while Bcl-2 gene expression and its protein expression levels were down-regulated compared with CK group.In hesperidin group,Bax and Caspase3 gene expression and their protein expression levels were significantly or extremely significantly down-regulated compared with CK,Bcl-2 gene expression and its protein expression levels were up-regulated compared with CK group.Compared with H2O2 group,Bax and Caspase3 gene expression and their proteins expression in H2O2+hesperidin group were extremely significantly down-regulated,and Bcl-2 gene expression and its protein expression were extremely significantly up-regulated.[Conclusion]Hesperidin inhibits H2O2-induced apop-tosis of bovine mammary epithelial cells and effectively enhances cell viability through Bcl-2/Bax-Caspase3 pathway,it is speculated that hesperidin has a good positive effect on promoting mammary organ development and maintaining lacta-tion abilityin dairy cow.
关键词
奶牛/橙皮苷/乳腺上皮细胞/细胞凋亡/Bcl-2/Bax-Caspase3信号通路
Key words
dairy cow/hesperidin/mammary epithelial cell/apoptosis/Bcl-2/Bax-Caspase3 signal pathway
维普
万方数据