脂多糖处理时间对山羊瘤胃上皮细胞损伤的影响

Anti-damage effects of goat rumen epithelial cells induced by lipopolysaccharide

在山羊瘤胃上皮细胞(GRECs)基础培养基中加入1μg/mL脂多糖(LPS),培养3、6、9 h后,检测细胞活性、抗氧化指标、炎症因子及紧密连接蛋白基因的表达。结果发现:1)LPS处理3 h组GRECs的活性显著低于处理6 h和9 h组的,而处理6 h和9 h组GRECs的活性无显著差异;2)LPS处理3 h组GRECs的谷胱甘肽过氧化物酶(GSH–PX)活性极显著低于处理6 h和9 h组的,但MDA含量的变化则相反,GRECs的SOD和CAT活性随LPS处理时间的延长而显著升高,ROS含量则随LPS处理时间的延长而极显著降低;3)LPS处理3h组GRECs的TNF–ɑ和IL–1β相对表达量极显著高于处理6h和9h组的,IL–1β相对表达量随LPS处理时间的延长而显著降低,各组间IL–6相对表达量无显著差异;4)LPS处理3 h组GRECs的ZO–1分布低,随着处理时间延长ZO–1分布增加,LPS处理3 h的GRECs紧密连接蛋白Claudin–1相对表达量显著高于处理6 h和9 h组的。说明随着LPS处理时间的延长,山羊瘤胃上皮细胞能够通过提高自身抗氧化和抗炎症能力来缓解氧化损伤,进而改善细胞屏障功能。

Goat rumen epithelial cells(GRECs)were cultured in medium containing 1μg/mL of LPS for 3 h,6 h and 9 h to detect cell viability,antioxidant indices,genes expression of inflammatory factors and tight junction protein.The results showed as follows:1)The viability of GRECs in the group treated with LPS for 3 h was significantly lower than that in groups treated with LPS for 6 h and 9 h,but there was no significant difference in GRECs activity between groups treated with LPS for 6 h and 9 h.2)The glutathione peroxidase(GSH-PX)activity of GRECs treated with LPS for 3 h was extremely significantly lower than that in cells treated with LPS for 6 h and 9 h(P<0.01),whereas the malondialdehyde(MDA)content,on the contrary,was extremely significantly higher in cells treated with LPS for 3 h.The superoxide dismutase(SOD)and catalase(CAT)activities of GRECs significantly increased and the reactive oxygen(ROS)concentration extremely decreased with the prolongation of LPS treatment time.3)The relative expression levels of TNF-ɑand IL-1βin GRECs treated with LPS for 3 h were significantly higher than that in cells treated with LPS for 6 h and 9 h.The relative expression of IL-1βwas significantly decreased with the extension of LPS treatment time.The relative expression levels of IL-6 had no significant difference among all group.4)The distribution of ZO-1 was low in GRECs treated with LPS for 3 h,but increased with the extension of LPS treatment time.The relative expression levels of Claudin-1 in GRECs treated with LPS for 3 h were significantly higher than that in cells treated with LPS for 6 h and 9 h.The results showed that the GRECs could alleviate oxidative damage by improving their own antioxidant and anti-inflammatory capacities with the extension of LPS treatment time,then improve barrier function of the cells.