摘要
目的探讨白细胞介素-22(IL-22)对肠上皮屏障紧密连接的影响及机制。方法将人结肠癌细胞系HT29分为对照组、IL-22组(100 ng/mL)、FLLL32组(IL-22干预前使用FLLL32预处理)、colivelin组(IL-22干预前使用colivelin预处理)。转染处理组:shNC组、shNC+IL-22组(IL-22干预前转染阴性对照)、shSTAT3组(转染LV-STAT3-RNAi)、shSTAT3+IL-22组(IL-22干预前转染LV-STAT3-RNAi)。用Western blot、real-time PCR检测HT29细胞STAT3、p-STAT3和紧密连接蛋白ZO-1、occludin、claudin-1和claudin-2及其mRNA的表达;用透射电镜观察HT29细胞紧密连接结构。结果相比于对照组,IL-22组的STAT3、p-STAT3、ZO-1、occludin、claudin-1和claudin-2蛋白及mRNA表达水平明显升高(P<0.05),且紧密连接更致密。相比于IL-22组,FLLL32组中上述指标的蛋白及mRNA表达量明显下调(P<0.05),紧密连接较差,colivelin组则表达上调(P<0.05),紧密连接完好。相比于shNC组,shNC+IL-22组的所有紧密连接的蛋白及mRNA表达量明显增加(P<0.05),紧密连接结构更致密;而相比于shSTAT3组,shSTAT3+IL-22组的所有紧密连接的蛋白及mRNA表达量的差异无统计学意义。结论STAT3通路参与IL-22促进HT29细胞紧密连接蛋白ZO-1、occludin、claudin-1和claudin-2的表达。
Objective To investigate the effect of interleukin-22(IL-22)on intestinal epithelial barrier tight junction and its mechanism.Methods Human colon cancer HT29 cells were divided into control group,IL-22 group(100 ng/mL),FLLL32 group(pretreatment with FLLL32 before IL-22 intervention)and colivelin group(pretreatment with colivelin before IL-22 intervention).The transfection treatment groups included:shNC group,shNC+IL-22 group(infection with negative control virus before IL-22 intervention),shSTAT3 group(infection with LV-STAT3-RNAi virus),and shSTAT3+IL-22 group(LV-STAT3-RNAi virus infection before IL-22 intervention).Western blot and real-time PCR were used to detect the protein and mRNA expression of STAT3,p-STAT3 tight junction protein ZO-1,occludin,claudin-1 and claudin-2 in HT29 cells.Transmission electron microscopy(TEM)was used to observe the changes in the structure of tight junctions of HT29 cells.Results The protein and mRNA expression level of STAT3,p-STAT3,ZO-1,occludin,claudin-1 and claudin-2 in IL-22 group were significantly higher than those in the control group(P<0.05)with more compact,tight junction.Compared with IL-22 group,protein and mRNA expression level of the above indexes in FLLL32 group were significantly decreased(P<0.05),the tight junction was poor,the expression of colivelin group was up-regulated(P<0.05),the tight junction was intact.The shNC+IL-22 group significantly increased the expression of all tightly bound proteins and mRNA compared with the shNC group(P<0.05),the tight junction structure was more dense,while the shSTAT3+IL-22 group showed no significant difference in protein and mRNA expression level when compared with theose of shSTAT3 group.Conclusions STAT3 signaling pathway is involved in IL-22 to promote the expression of tight junction proteins ZO-1,occludin,claudin-1 and claudin-2 in HT29 cells.
作者
宋茜
黄雪
覃蒙斌
张君红
井洁
罗毅华
李雨芯
SONG Qian;HUANG Xue;QIN Meng-bin;ZHANG Jun-hong;JING Jie;LUO Yi-hua;LI Yu-xin(Department of Geriatric Gastroenterology,the First Affiliated Hospital,Guangxi Medical University,Nanning 530021;Department of Gastroenterology,the Second Affiliated Hospital,Guangxi Medical University,Nanning 530000,China)
出处
《基础医学与临床》
2021年第6期792-798,共7页
Basic and Clinical Medicine
基金
国家自然科学基金(81660093)
广西研究生教育创新计划(YCSW2020112)。
