血必净注射液对脓毒症大鼠Claudin4蛋白和辅助性T淋巴细胞1型表达的影响  

Effect of Xuebijinginjection on Claudin4 protein and T helper cell type1 in rats with sepsis

作　　者：刘梅[1,2] 邓星奇 李文放[1] 林兆奋[1] 凌美蓉 

机构地区：[1]上海长征医院急救科,上海200003 [2]上海市闵行区中心医院急救科 [3]上海市闵行区中心医院急诊科

出　　处：《上海医学》2015年第2期133-138,I0003,共7页Shanghai Medical Journal

基　　金：上海市医学重点专科建设计划项目资助(ZK2012A23)

摘　　要：目的探讨血必净注射液对脓毒症大鼠肺组织Claudin4蛋白表达的影响。方法健康成年Sprague-Dawley大鼠100只,体重(250±25)g,随机分入4组,每组25只:假手术组大鼠只开腹、关腹和复苏,不行盲肠结扎穿孔术(CLP);脓毒症组大鼠行CLP;血必净3h组大鼠于CLP后3h腹腔内注射血必净注射液2mL/kg;血必净12h组大鼠于CLP后12h腹腔内注射血必净注射液2mL/kg。每组随机留取8只大鼠进行生存率分析。假手术组、脓毒症组、血必净3h组大鼠在术后6、15和24h,血必净12h组大鼠在术后15和24h各随机处死5只,分别留取血液和肺组织标本,测定肺组织湿/干质量比(肺水肿指数),对肺组织进行病理学观察并进行肺损伤病理学评分,测定血清促炎细胞因子IL-6和TNF-α水平,采用免疫组织化学方法测定肺组织骨架蛋白Claudin4蛋白表达情况,采用流式细胞术检测辅助性T淋巴细胞1型(Th1)在外周血单核细胞中的表达情况。结果假手术组、脓毒症组、血必净3h组和血必净12h组大鼠的术后72h存活率分别为8/8、0、1/8和0,血必净3h组显著高于脓毒症组(P<0.01),血必净12h组与脓毒症组的差异无统计学意义(P>0.05)。假手术组大鼠术后各项检测指标均基本不变。脓毒症组、血必净3h组和血必净12h组大鼠术后15和24h的肺水肿指数,以及术后6、15和24h的肺损伤病理学评分、血清IL-6和TNF-α水平、肺组织Claudin4蛋白表达阳性率均显著高于假手术组同时间点(P值均<0.01);脓毒症组大鼠术后6、15和24h,以及血必净12h组大鼠术后15和24h的肺水肿指数、肺损伤病理学评分、血清IL-6和TNF-α水平、肺组织Claudin4蛋白表达阳性率均显著高于血必净3h组同时间点(P值分别<0.05、0.01);脓毒症组与血必净12h组术后同时间点间上述指标的差异均无统计学意义(P值均>0.05)。脓毒症组和血必净3h组大鼠术后各时间点的外周血单核细胞中Th1的表达率均显著高于假手术�Objective To investigate the protective effects of Xuebijing injection on cytoskeletal protein Claudin4 protein expression in septic rats and the underlying molecular mechanisms. Methods A total of 100 healthy adult Sprague-Dawley rats （weighing [250± 251 g） were randomly divided into sham operation group （NS group, n =25）, sepsis group （cecal ligation and puncture [-CIP]group, n = 25）, Xuebijing was given 3 hours after CLP （Xuebijing 3 h group, n = 25）, and Xuebijing was given 12 hours after OLP （Xuebijing 12 h group, n = 25）. Eight rats were randomly sampled in each group for survival analysis. Rats in NS group, sepsis group, and Xuebijing 3 h group were sacrificed at 6, 15 and 24 h after operation, and those in the Xuebijing 12 h group were sacrificed at 15 and 24 hours after operation. The blood samples and lung tissues of rats were collected; Wet-to- dry lung weight ratio and pathologic changes of lungs were observed to evaluate pathological scores of lungs. The serum tumor necrosis factor （TNF）-α and interleukin （IL）-6 levels were examined in the serum samples. The expression of Claudin4 in lung tissues was examined by immunohistochemistry technology, and the proportion of T helper cell type 1 （Th1） cells in peripheral blood cells was analyzed by flow cytometry. Results The survival rates of NS group, CLP group, Xuebijing 3 h group and Xuebijing 12 h group were 8/8, 0, 1/8 and 0 at 72 h after operation, respectively, with that of Xuebijing 3 h group being significantly higher than that of the sepses group （P〈0. 01） and that of Xuebijing 12 h group being similar to that of sepses group （P〉0. 05）. The parameters in the sham operation group basically kept unchanged after operation. The pulmonary edema indexes of CLP group, Xuebijing 3 h group and Xuebijing 12 h group at 15 and 24 h after operation were significantly higher than that of NS group at the same time points （all P〈0. 01 ）. The pathological scores, levels of TNF-α, IL-6 of CLP group, Xuebijing 3 h

关 键 词：脓毒症 肺损伤 血必净 促炎细胞因子 Claudin4蛋白 辅助性T淋巴细胞1型 

分 类 号：R459.7[医药卫生—急诊医学]