脂多糖对奶牛子宫内膜上皮细胞氧化损伤的影响机制

Effects and mechanism of lipopolysaccharide on oxidative damage of bovine endometrial epithelial cells

为探究脂多糖(LPS)对原代奶牛子宫内膜上皮细胞(BEEC)氧化损伤的影响机制,本研究采用1μg/m L LPS处理细胞,利用流式细胞术和试剂盒在不同时间点检测活性氧水平、超氧化物歧化酶(SOD)活力和丙二醛(MDA)含量的变化;利用荧光定量PCR和Western-blot技术检测NRF2/KEAP1通路相关基因和蛋白表达的时程变化规律。结果显示,LPS处理BEEC细胞12 h可引起细胞活性氧水平和MDA含量增加(P<0.01)、SOD活力降低(P<0.05);LPS处理BEEC细胞6、12和24 h,细胞NRF2、HMOX1和NQO1的m RNA丰度降低(P<0.05);LPS作用于BEEC细胞90 min,细胞NRF2/KEAP1通路关键蛋白表达下降(P<0.05)。表明,LPS可导致原代BEEC细胞氧化损伤并抑制NRF2/KEAP1通路激活。

To explore the mechanism of lipopolysaccharide(LPS) on oxidative damage in primary bovine endometrial epithelial cells(BEEC),the present study detected the level of reactive oxygen species by flow cytometry,and the activity of superoxide dismutase(SOD) and the content of malondialdehyde(MDA) by kit at different time points.The mRNA abundance of related genes and the level of key proteins in NRF2/KEAP1 pathway were detected by quantitative PCR and Western-blot,respectively at different time points.In results,the level of reactive oxygen species and the content of MDA increased(P<0.01) and the activity of SOD decreased(P<0.05) after LPS treatment for 12 h.The mR NA abundance of NRF2,HMOX1 and NQO1 decreased(P<0.05) after LPS-treatment for 6,12,and 24 h.The protein expression of NRF2/KEAP1 pathway decreased after LPS-treatment for 90 min(P<0.05).In conclusion,LPS induced oxidative damage and inhibited the activation of NRF2/KEAP1 pathway in primary BEEC.