内质网应激诱导的自噬对新生大鼠坏死性小肠结肠炎的影响  

Effect of Endoplasmic Reticulum Stress-induced Autophagy on Necrotizing Enterocolitis in Neonatal Rats

作　　者：王丽红[1] 杨晓丽[2] 李静[2] WANG Lihong;YANG Xiaoli;LI Jing(Shanxi People′s Hospital,Shanxi 030000,China)

机构地区：[1]山西医科大学儿科医学系,太原030000 [2]山西省人民医院儿科,太原036200

出　　处：《医学研究杂志》2024年第2期106-111,共6页Journal of Medical Research

基　　金：山西省卫生健康委员会科研课题(2020024)。

摘　　要：目的探讨抑制内质网应激(endoplasmic reticulum stress,ERS)诱导的自噬对新生大鼠坏死性小肠结肠炎(necrotizing enterocolitis,NEC)的影响。方法首先建立新生大鼠NEC模型,然后从中分离提取出肠上皮细胞,分为对照组、抑制组、诱导组。对照组正常培养,抑制组加4-苯基丁酸,诱导组加衣霉素处理24h。采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)检测各组细胞炎性细胞因子肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、肠脂酸结合蛋白(intestinal fatty acid binding protein,I-FABP)的表达水平;实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)检测各组细胞ERS标志物葡萄糖调节蛋白78(glucose regulated protein 78,GRP78)、氧调节蛋白150(oxygen-regulated protein 150,ORP150)的mRNA表达水平;Western blot法检测各组细胞自噬相关蛋白LC3Ⅱ/Ⅰ、p62的表达水平。结果与对照组比较,抑制组p62表达明显增高,TNF-α、I-FABP、GRP78、ORP150、LC3Ⅱ/Ⅰ表达显著降低,而诱导组p62表达明显降低,TNF-α、I-FABP、GRP78、ORP150、LC3Ⅱ/Ⅰ表达显著增高,差异均有统计学意义(P<0.05)。结论抑制ERS诱导的自噬激活可减轻NEC新生大鼠肠黏膜损伤和炎性反应,改善肠道屏障功能。Objective To investigate the effect of inhibiting autophagy induced by endoplasmic reticulum stress(ERS)on necrotizing enterocolitis(NEC)in neonatal rats.Methods First,the NEC model of neonatal rats was established.Then,the intestinal epithelial cells were isolated and divided into three groups:control group,inhibition group and induction group.The control group was cultured normally,the inhibition group was added with 4-phenylbutyric acid,and the induction group was added with tunicamycin for 24hours.Enzyme-linked immunosorbent assay(ELISA)was used to detect the expression of the cellular inflammatory cytokines tumor necrosis factor-α(TNF-α)and intestinal fatty acid binding protein(I-FABP)in each group.Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the mRNA expression level of the markers of ERS glucose regulated protein 78(GRP78)and oxygen-regulated protein 150(ORP150).Western blot was used to detect the expression of autophagy related proteins LC3Ⅱ/Ⅰand p62.Results Compared with the control group,the expression of p62 in the inhibition group increased significantly,the expression of TNF-α,I-FABP,GRP78,ORP150,LC3Ⅱ/Ⅰin the inhibition group was significantly decreased,while the expression of p62 in the induction group was significantly decreased,the expressions of TNF-α,I-FABP,GRP78,ORP150,LC3Ⅱ/Ⅰwere significantly increased,and the differences were statistically significant(P<0.05).Conclusion Inhibition of ERS induced autophagy activation can alleviate intestinal mucosal injury and inflammatory response in neonatal rats with NEC and improve intestinal barrier function.

关 键 词：内质网应激 自噬 坏死性小肠结肠炎 肠上皮细胞 作用机制 

分 类 号：R722[医药卫生—儿科]