线粒体靶向抗氧化肽SS-31对脓毒症大鼠血管通透性的保护作用  

Mitochondria-targeted peptide SS-31 protects septic rats against vascular permeability

作　　者：朱娱[1] 吴跃[1] 张杰[1] 田昆仑[1] 彭小勇[1] 向鑫明 刘良明[1] 李涛[1] ZHU Yu;WU Yue;ZHANG Jie;TIAN Kunlun;PENG Xiaoyong;XIANG Xinming;LIU Liangming;LI Tao(State Key Laboratory of Trauma,Burns and Combined Injury,Department 2,Institute of Surgery Researeh,'Third Affiliated Hospital,Army Medical University (Third Military Medical University),Chongqing,400042,China)

机构地区：[1]陆军军医大学(第三军医大学)第三附属医院(野战外科研究所)第二研究室,创伤,烧伤与复合伤国家重点实验室,重庆400042

出　　处：《第三军医大学学报》2019年第3期199-205,共7页Journal of Third Military Medical University

基　　金：国家自然科学基金青年科学基金(81701897)~~

摘　　要：目的探讨线粒体靶向抗氧化肽SS-31对脓毒症大鼠血管通透性的影响及其机制。方法40只12周龄SD大鼠(雌雄各半,体质量约220 g)分成5组(n=8):假手术组(只开腹,不结扎盲肠和穿孔)、脓毒症组(盲肠结扎穿孔术制作脓毒症模型)、常规治疗组(conventional treatment,CT组,脓毒症模型12 h后,股静脉输注LR)、SS-31早期治疗组(常规治疗基础上在盲肠结扎穿孔前30 min尾静脉给予SS-31 3 mg/kg)和SS-31晚期治疗组(常规治疗基础上在盲肠结扎穿孔12 h后从股静脉输注LR联合SS-31)。通过荧光蛋白透过率的方法测定大鼠肺脏和肾脏的血管通透性,伊文思蓝染色透过率检测大鼠肠道通透性;离体取原代肺静脉内皮细胞观察LPS刺激(LPS组)及SS-31预处理(SS-31预处理组)后对内皮细胞的闭锁小带蛋白1(zonula occludes-1,ZO-1)的表达和细胞跨膜电阻(trans electric resistance,TER),以及活性氧(reactive oxygen species,ROS)的影响,以无血清的基础培养基培养24 h的细胞作为正常对照组。结果 (1)与假手术组比较,脓毒症组大鼠肺脏、肾脏血管以及肠道组织的通透性明显增加,线粒体功能降低(P <0. 01);采用常规治疗(LR联合头孢呋辛钠和多巴胺同时输注)尽管可以降低脓毒症大鼠的通透性和减轻线粒体功能损害,但与脓毒症组比较,差异无统计学意义(P>0. 05); SS-31治疗可明显降低脓毒症大鼠肺脏、肾脏血管以及肠道组织的通透性和增加线粒体功能,与常规治疗组比较,差异有统计学意义(P <0. 01),其中SS-31早期治疗效果比晚期治疗效果更好。(2)与正常对照组比较,LPS组ZO-1表达明显降低,细胞结构排列紊乱,TER明显降低,ROS含量增加; SS-31预处理可使ZO-1表达增加,细胞间连接紧密,TER明显增加,ROS含量明显降低。结论 SS-31对脓毒症大鼠的血管通透性有保护作用,其机制可能与抑制线粒体氧化应激有关。Objective To investigate the protective effect of mitochondria-targeted peptide SS-31 on vascular permeability in septic rats and explore its underlying mechanism. Methods Forty Sprague-Dawley rats( 12 weeks old,both genders,weighting about 220 g) were equally and randomly divided into 5 groups,sham operation group( sham group),sepsis group,sepsis + conventional treatment( CT) group,sepsis +early injection of SS-31 group and sepsis + later injection of SS-31 group. Cecal ligation and puncture( CLP)was performed to establish septic rats. Lactated Ringer ’s solution( containing cefuroxime sodium and dopamine) or the solution containing 3 mg/kg SS-31 were injected through femoral vein to the rats of sepsis +CT group or sepsis + later injection group,and the agent was also given to the rats of the sepsis + early injection group in 30 min before CLP. Fluorescence albumin transmissivity in the tissue homogenate was measured for the vascular permeability of the lungs and kidneys. Intestinal permeability was measured by Evans blue staining. Primarily isolated and cultured pulmonary vein endothelial cells were treated with lipopolysaccharide( LPS group),and SS-31 followed by LPS addition( LPS + SS-31 group). Then the expression level of zonula occludes-1( ZO-1),intensity of transmembrane resistance( TER),and generation of reactive oxygen species( ROS) were detected in the endothelial cells. The cells cultured in serum-free medium for 24 h served as normal control cells. Results(1) The sepsis group had significantly more obvious vascular permeability of the lungs,kidneys and intestine,and damaged mitochondrial function when compared with the sham group( P < 0. 01). Though conventional treatment reduced the permeability and relieved mitochondrial dysfunction,the effects were not significant than the conditions in the sepsis group( P > 0. 05).Both early and later injection of SS-31 notably decreased the permeability and improved mitochondrial function than the conventional treatment( P < 0. 01),with the effects of early in

关 键 词：线粒体靶向抗氧化肽SS-31 脓毒症大鼠 氧化应激 通透性 

分 类 号：R543[医药卫生—心血管疾病] R631.1[医药卫生—内科学]