血必净对创伤弧菌脓毒症大鼠肺组织核因子-KBp65表达作用的研究  

Expression of nuclear factor - kB p65 in the lung of the rat with vibrio vulnificus sepsis and intervention with Xuebijing injection

作　　者：李忠旺[1] 洪广亮[1] 孙琦[1] 卢中秋[1] 邱俏檬[1] 梁欢[1] 

机构地区：[1]温州医学院附属第一医院急诊科,浙江325000

出　　处：《中国急救医学》2010年第10期905-909,961,共6页Chinese Journal of Critical Care Medicine

基　　金：基金项目：浙江省医学扶植重点建设学科计划项目（No.07-1704）;浙江省中医药科技计划项目（No.2007CA085）;温州市科技计划项目（No.Y20080088）

摘　　要：目的 观察创伤弧菌脓毒症大鼠肺组织核因子-κB（NF-κB）p65基因及蛋白表达,并探讨血必净对其的干预作用.方法 SD大鼠110只,随机分为正常组（A组,n＝10）、创伤弧菌脓毒症组（B组,n＝50,采用大鼠左下肢皮下注射创伤弧菌悬液制作大鼠创伤弧菌脓毒症模型）和血必净治疗干预组（C组,n＝50,感染后半小时腹腔注射血必净4 mL/kg）.B、C组于染菌后1、6、12、24、48 h活杀（各时间点n=10）,采用逆转录-聚合酶链式反应（RT-PCR） 法、Western blot法和双抗体夹心酶联免疫吸附法（ELISA）分别检测大鼠肺组织NF-κB p65的基因与蛋白表达及IL-10的含量,数据采用单因素方差分析.结果 与A组比较,B组大鼠肺组织创伤弧菌感染后6、12 h NF-κB p65 mRNA表达量与6、12、24和48 h肺组织NF-κB p65蛋白表达量均明显增高 （P〈0.05）;与B组相同时间点比较,C组6 h肺组织NF-κB p65 mRNA表达量与12、24及48 h肺组织NF-κB p65 蛋白表达量明显减少（P〈0.05）; 与A组比较,B组创伤弧菌菌感染12、24和48 h IL-10的含量明显增加（P〈0.05）,与B组相同时间点比较,C组24 h（52.444±9.605）肺组织IL-10的含量明显增高（P〈0.05）;感染后48 h,大鼠肺内血管明显充血,间质水肿并伴炎性浸润,肺泡腔塌陷,血必净干预后,肺组织损伤有所减轻.结论 NF-κB参与了创伤弧菌脓毒症肺损伤过程;血必净能抑制NF-κB p65的表达,从而起到保护创伤弧菌脓毒症大鼠肺组织的作用.Objective To observe the expression of NF - KB p65 gene and protein in the lung of rats with vibrio vulnificus sepsis, and investigate the intervention of Xuebijing injection. Methods one hundred and ten rats of clean grade were randomly divided into normal control group （ A group, n = 10）, vibrio vulnificus sepsis group （B group, n =50, sepsis was reproduced in rats with subcutaneous injection in left lower limb with vibrio vulnifieus） and the intervention of Xuebijing treatment group （C group, n =50, rats were intraperitoneal（ip） with tile dose of Xuebijing 4 mL/kg at the time of 30 min later after infection）. The rats in group B and C were sacrificed at 1, 6, 12, 24, 48 h after infection. Respectively, reverse transcription -polymerase chain reaction （RT -PCR）, Western blot and double - antibody sandwich enzyme - linked immunosorbent assay （ELISA） were used to detect NF - KB p65 gene, protein and IL - 10 in rat lung. Results Compared with group A , after infected 6 h and 12 h NF - KB p65 mRNA and 6 h, 12 h ,24 h,48 h NF - KB p65 protein expressions in lung of the rats in group B were significantly higher（P 〈0. 05） ;compared with group B, after infected 6 h NF- KB p65 mRNA and 12 h, 24 h, 48 h NF - KB p65 protein expressions in lung of the rats in group C were significantly lower （ all P 〈 0. 05 ）. Compared with group A, the level of IL - 10 in group B rat lung significantly increased after infection at 12 h,24 h and 48 h（ all P 〈 0. 05 ） ; Compared with group B, the level of IL - 10 in the lung of rats in group C significantly increased at 24 h（P 〈0. 05） ： Compared with group A, the histopathological of the lung of group B changed markedly at 48 h, which showed pulmonary vascular congestion, hemorrhage, interstitial edema, inflammatory infiltration and alveolar cavity collapse; Compared with the same time point of group B, the lung of group C showed some relief. Conclusion NF - KB was involved in the vibrio vulnificus infection in the pathogenesis of acute lung

关 键 词：创伤弧菌 脓毒症 核因子-KB p65(NF-KB N5) 白细胞介素-10(IL-10) 血必净注射液 

分 类 号：R758.630.2[医药卫生—皮肤病学与性病学]