高压氧对缺血再灌注肠黏膜上皮Caco-2细胞间紧密连接的影响  

Effects of hyperbaric oxygen on the tight junction of intestinal epithelial cells Caco-2 in ischemia-reperfusion injury

作　　者：冯维喆 褚海涛[1] 王勇[1] 王钢[1] 

机构地区：[1]中国医科大学附属第一医院急诊科,沈阳110001

出　　处：《中华航海医学与高气压医学杂志》2015年第2期105-109,共5页Chinese Journal of Nautical Medicine and Hyperbaric Medicine

基　　金：辽宁省自然科学基金项目（20092137）

摘　　要：目的 观察高压氧（HBO）对缺血再灌注（I/R）人类肠黏膜上皮Caco-2细胞间紧密连接（TJ）的影响.方法 培养单层人类大肠黏膜上皮细胞Caco-2,应用0.5 mmol/L丁基过氧化氢（t-buooh）诱导离体肠黏膜上皮细胞I/R损伤.按数字表法随机分为5组（每组8个培养孔）：对照组（N组）、I/R组、HBO预处理（HBO-P）组、缺血期HBO干预（HBO-I）组、再灌注期HBO干预（HBO-R）组.采用跨膜电阻抗检测法（TEER）检测肠道黏膜上皮细胞间紧密连接的开放程度并评价其通透性.用Rt-PCR检测细胞间紧密连接蛋白ZO-1转录水平.结果 TEER阻值N组最高,其次为HBO-P组,再次为HBO-R组,最后为HBO-I组.N组与其他各组差异均有统计学意义（P＜0.01）;HBO-P组阻值与I/R组、HBO-I组及HBO-R组的阻值比较,差异均有统计学意义（P＜0.05）;HBO-R组与I/R组、HBO-I组阻值比较差异有统计学意义（P＜0.05）;L/R组高于HBO-I组,且差异有统计学意义（P＜0.05）.ZO-I mRNA的Rt-PCR结果显示：与N组相比,其他各组ZO-1 mRNA含量均明显下降,且差异有统计学意义（P＜0.01）;HBO-P组ZO-1 mRNA含量均高于L/R组、HBO-R组及HBO-I组（P＜0.01）;HBO-R组ZO-1 mRNA含量高于I/R组和HBO-I组（P＜0.05）;HBO-I组与I/R组ZO-1 mRNA含量接近,差异无统计学意义（P＞0.05）.结论 HBO干预具有保护单层Caco-2细胞TJ功能作用,抑制TJ开放,机制可能与维持TJ的骨架蛋白ZO-1有关.缺血期前和再灌注期给予HBO干预治疗显示有益结果,HBO预处理疗效最佳,缺血期给予HBO无效.Objective To investigate the effects of hyperbaric oxygen （HBO） on the tight junction （T J） of monolayer human intestinal epithelial cells Caco-2 in ischemia reperfusion （I/R） injury in vitro.Methods In the culture of human intestinal monolayer epithelial cells Caco-2,0.5 mmol/L t-buooh was used to induce I/R injury of intestinal epithelial cells in vitro.The monolayer Caco-2 cells were randomly divided into 5 groups （n =8 cases）：the control group,the ischemia/reperfusion group （I/R）,the hyperbaric oxygen preconditioning group （HBO-P）,the HBO intervention during ischemia group （HBO-I） and the HBO intervention during reperfusion group （HBO-R）.Transmembrane electrical impedance detection （TEER） was used to detect the opening of the tight junction between epithelial cells of the intestinal mucosa and evaluate its permeability.The transcription of tight ligandin ZO-1 between cells was detected by RT-PCR.Results Results indicated that the impedance as shown by TEER was in the following order：the impedance value of the control group was the highest,then came the HBO-P group,further down was the HBO-R group,and at the end was the HBO-I group.Statistical significance could be noted when comparisons were made between the HBO-Pgroup and all the other groups（P ＜ 0.01）.There was also statistical significance,when the impedance value of the HBO-P group was compared with those of the I/R,HBO-I and HBO-R groups （P ＜ 0.05）.Statistical significance could also be noted,when the impedance value of the HBO-R group was compared with those of the I/R and HBO-I groups（P ＜0.05）.The impedance value of the I/R group was higher than that of the HBO-I group,with statistical significance（P ＜0.05）.RT-PCR results of ZO-1 indicated that the ZO-1 mRNA levels of the I/R,the HBO-P,the HBO-R and the HBO-I groups were decreased significantly,as compared with that of the control group,with statistical significance （P ＜ 0.01）.The ZO-1 mRNA level of the HBO-P group was higher than th

关 键 词：高压氧 缺血/再灌注损伤 紧密连接 CACO-2细胞 ZO-1 

分 类 号：R459.7[医药卫生—急诊医学]