氢对脓毒症小鼠肠组织ROCK1和mDia1表达的影响  

Effects of hydrogen on expression of ROCK1 and mDial in intestinal tissues of septic mice

作　　者：张红涛[1] 姜连浩[1] 刘玲玲[1] 张素品[2] 梁禹[1] 于泳浩[2] 

机构地区：[1]天津市环湖医院麻醉科,300350 [2]天津医科大学总医院麻醉科天津市麻醉学研究所,300052

出　　处：《中华麻醉学杂志》2017年第3期356-360,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（81071533,81372033）;天津市应用基础及前沿技术研究计划（11JCYBJC12900）

摘　　要：目的评价氢对脓毒症小鼠肠组织Rho相关激酶1（ROCK1）和哺乳动物Dia相关蛋白1（mDial）表达的影响。方法雄性C57BL／6小鼠90只，体重20～25g，6周龄，采用随机数表法，将其分为3组（n=30）：对照组（c组）、脓毒症组（s组）和脓毒症＋氢组（SH组）。采用盲肠结扎穿孔（CLP）法制备脓毒症模型。SH组于CLP后1和6h时吸入2％氢气1h。取20只小鼠观察7d生存情况；于CLP术后24h时处死10只小鼠，心脏取血，测定血清二胺氧化酶（DAO）活性，并行血液细菌培养并计数菌落数。取小肠组织，行肠损伤评分，采用免疫组织化学染色法测定ROCK1和mDial阳性细胞率，并计算ROCK1与mDial阳性细胞率的比值（ROCK1／mDial比值）；采用免疫荧光染色法测定肠上皮细胞钙粘附蛋白E的表达。结果与c组比较，S组和SH组生存率降低，血清DAO活性、血液培养菌落数及肠损伤评分升高，肠组织ROCK1阳性细胞率和mDial阳性细胞率升高，钙粘附蛋白E表达下调，s组ROCK1／mDial比值升高（P〈0．05），SH组ROCK1／mDial比值差异无统计学意义（P〉0．05）；与s组比较，SH组生存率升高，血清DAO活性、血液培养菌落数及肠损伤评分降低，肠组织ROCK1阳性细胞率降低，mDial阳性细胞率升高，ROCK1／mDial比值降低，钙粘附蛋白E表达上调（P〈0．05）。结论氢改善脓毒症小鼠肠屏障功能的机制可能与下调ROCK1表达，上调mDial表达，纠正ROCK1，mDial比例失衡有关。Objective To evaluate the effects of hydrogen on the expression of Rho-associated pro- tein kinase 1 （ROCK1） and mammalian diaphanous-related formin 1 （mDial） in intestinal tissues of sep- tic mice. Methods Ninety male C57BL/6 mice, weighing 20-25 g, aged 6 weeks, were divided into 3 groups （n= 30 each） using a random number table： control group （group C）, sepsis group （group S） and sepsis plus hydrogen group （group SH）. Sepsis was produced by cecal ligation and puncture （CLP）. Group SH inhaled 2% hydrogen for 1 h starting from 1 and 6 h after CLP. Twenty mice in each group were selected and observed for 7-day survival rate. Ten mice in each group were sacrificed at 24 h after CLP, and blood samples were obtained from hearts to measure the activity of serum diamine oxidase （DAO） and to count the colony-forming units after bacterial culture. The small intestinal samples were obtained for mi- croscopic examination of pathological changes and for determination of ROCK1 and mDial positive cell rates （using immunohistochemical staining） and expression of intestinal epithelial junctional protein E-cadherin （by immunofluorescent staining）. Intestinal damage was assessed and scored. The ratio of ROCK1 to mDial positive cell rates （ROCK1/mDial ratio） was calculated. Results Compared with group C, the survival rate was significantly decreased, the serum DAO activity, colony counts and intestinal damage scores were increased, ROCK1 and mDial positive cell rates were increased, and the expression of E-cad- herin was down-regulated in S and SH groups, ROCK1/mDial ratio was increased in group S （P〈0.05） , and no significant change was found in ROCK1/mDial ratio in group SH （P〉0.05）. Compared with group S, the survival rate was significantly increased, the serum DAO activity, colony counts and intestinal dam- age scores were decreased, the ROCK1 positive cell rate was decreased, the radial positive cell rate was increased, ROCK1/mDial ratio was decreased, and the express

关 键 词：氢 脓毒症 RHO相关激酶类 PA GTP结合蛋白质 肠粘膜 

分 类 号：R459.7[医药卫生—急诊医学]