摘要
目的探讨LINC00943对幽门螺杆菌(Hp)诱导的人胃黏膜上皮细胞GES-1损伤的影响及其分子机制。方法将GES-1细胞分为Control组、Hp组、Hp+si-NC组、Hp+si-LINC00943组、Hp+miR-NC组、Hp+miR-125a-5p组、Hp+si-LINC00943+anti-miR-NC组和Hp+si-LINC00943+anti-miR-125a-5p组,实时荧光定量PCR检测LINC00943和miR-125a-5p表达水平;流式细胞术检测细胞凋亡;免疫蛋白印迹检测凋亡蛋白表达;酶联免疫吸附法检测白细胞介素-8(IL-8)和IL-1β、乳酸脱氢酶(LDH)水平;双荧光素酶报告基因实验检测LINC00943和miR-125a-5p靶向关系。结果与Control组比较,Hp组LINC00943表达、凋亡率、Bax蛋白、IL-8、IL-1β、LDH升高,miR-125a-5p表达、Bcl-2蛋白降低(P<0.05);敲减LINC00943或过表达miR-125a-5p可减轻Hp诱导的GES-1细胞损伤;LINC00943靶向调控miR-125a-5p表达,下调miR-125a-5p逆转了敲减LINC00943对Hp诱导的GES-1细胞损伤的影响(P<0.05)。结论敲减LINC00943通过靶向上调miR-125a-5p可缓解Hp诱导的GES-1细胞损伤。
OBJECTIVE To explore the effect of LINC00943 on Helicobacter pylori-induced damage of human gastric mucosal epithelial cells GES-1 and observe its molecular mechanisms.METHODS The GES-1 cells were divided into the Control group,the Hp group,the Hp+si-NC group,the Hp+si-LINC00943 group,the Hp+miR-NC group,the Hp+miR-125a-5p group,the Hp+si-LINC00943+anti-miR-NC group and the Hp+si-LINC00943+anti-miR-125a-5p group.The expression levels of LINC00943 and miR-125a-5p were detected by real-time fluorescent quantitative PCR;the cell apoptosis wad detected by flow cytometry,the expression of apoptin was detected by Western blot,and the levels of interleukin-8(IL-8),IL-1βand lactic dehydrogenase(LDH)were detected by using enzyme-linked immunosorbent assay(ELISA).The targeted relationship between LINC00943 and miR-125a-5p was detected by means of dual luciferase reporter assay.RESULTS The expression level of LINC00943,apoptosis rate,Bax protein level,IL-8 level,IL-1βlevel and LDH level were higher in the Hp group than in the Control group,while the expression level of miR-125a-5p and Bcl-2 protein level were lower in the Hp group than in the Control group(P<0.05).The knockdown of LINC00943 or overexpression of miR-125a-5p could alleviate the Hp-induced damage of GES-1 cells.LINC00943 targeted regulation of miR-125a-5p and down-regulation of miR-125a-5p reversed the Hp-induced damage of GES-1 cells by the knockdown of LINC00943(P<0.05).CONCLUSION The knockdown of LINC00943 may alleviate the Hp-induced damage of GES-1 cells by targeted up-regulation of miR-125a-5p.
作者
朱丽君
李伟智
李游
唐瑜敏
雷青梅
ZHU Li-jun;LI Wei-zhi;LI You;TANG Yu-min;LEI Qing-mei(The First Affiliated Hospital of Jinan University,Guangzhou,Guangdong 5510000,China;不详)
出处
《中华医院感染学杂志》
CAS
CSCD
北大核心
2023年第2期198-202,共5页
Chinese Journal of Nosocomiology
基金
广东省医学科学研究基金资助项目(A2017007)。
