不同压力CO_2缺血预处理对气腹模型大鼠肝脏缺血再灌注损伤的影响  

Protective effects of ischemic preconditioning with different pressure of CO_2 on hepatic ischemia reperfusion injury in a rat model of CO_2 pneumoperitoneum

作　　者：陈贝贝[1] 杜敏[1] 王亮[1] 叶茂[1] 唐文[1] 

机构地区：[1]儿童发育疾病研究省部共建教育部重点实验室儿科学重庆市重点实验室重庆市儿童发育重大疾病诊治与预防国际科技合作基地重庆医科大学附属儿童医院麻醉科,400014

出　　处：《免疫学杂志》2015年第12期1030-1035,共6页Immunological Journal

基　　金：重庆市卫生局医学科学技术研究项目(2009-2-249);国家临床重点专科建设项目(国卫办医涵[2013]544)

摘　　要：目的探讨不同压力CO2缺血预处理对气腹模型大鼠肝脏缺血再灌注损伤的影响及可能机制。方法成年健康雄性SD大鼠30只,采用随机数字表法分为5组:空白对照组(C组)于麻醉后插入气腹针;气腹对照组(Pp组)以10 mm Hg气腹压力建立CO2气腹60 min;低压预处理组(L-IP组)、中压预处理组(M-IP组)、高压预处理组(H-IP组)在建立CO2气腹前,分别以5、10、15 mm Hg充气5 min放气5 min,连续3个循环。各组恢复灌注60 min后取肝组织,行HE染色光镜下观察肝组织病理学变化,分光光度法测定超氧化歧化酶(SOD)活性、总抗氧化物能力(T-AOC)及丙二醛(MDA)含量,免疫组化、Western blot法检测Bcl-2、Bax蛋白表达水平,RT-PCR法检测IL-1β、IL-6 m RNA表达水平。结果除C组外,Pp组与各IP组大鼠肝组织均出现病理学改变,其中,各IP组的损伤程度明显轻于Pp组;与Pp组比较,各IP组SOD活性、T-AOC升高及MDA含量下降,Bcl-2蛋白表达增加、Bax蛋白表达减少且Bcl-2/Bax比值升高,IL-1β及IL-6 m RNA表达下调(P均<0.05);与L-IP组、H-IP组比较,M-IP组SOD、TAOC、Bcl-2及Bcl-2/Bax比值均增加,MDA、Bax、IL-1β及IL-6 m RNA均减少。结论 CO2缺血预处理可减轻气腹模型大鼠肝脏缺血再灌注损伤,其中10 mm Hg压力的CO2预处理抗损伤作用更明显,其机制与减弱氧化应激反应,上调Bcl-2抗凋亡蛋白表达,下调Bax促凋亡蛋白表达及炎症细胞因子IL-1β、IL-6 m RNA的表达,减少细胞凋亡和降低炎性反应有关。This study aimed to investigate the protective effects of ischemic preconditioning with different pressure of CO2 on hepatic ischemia reperfusion injury in a rat model of CO2 pneumoperitoneum. Thirty healthy male Sprague-Dawley rats were randomly divided into five groups： control group（C group） was subjected to sham operation, pneumoperitoneum group（Pp group） was subjected to 60 minutes of pneumoperitoneum with 10 mm Hg of pressure, and three ischemic preconditioning groups（IP groups） were subjected to ischemic preconditioning prior to the induction of pneumoperitoneum. Ischemic preconditioning was defined as three cycles of 5 minutes of pneumoperitoneum with 5 mm Hg（L-IP group）, 10 mm Hg（M-IP group） or 15 mm Hg（H-IP group） intra-abdominal pressure, followed immediately by 5 minutes of deflation. Sixty minutes after deflation, hepatic tissues were collected at the end of the experiment to observe the pathological changes with HE staining; malondialdehyde（MDA）, superoxide dismutase（SOD） and total antioxidant capacity（T-AOC） were measured by spectrophotometry;protein expressions of Bcl-2 and Bax in the hepatic tissues were detected by immunohistochemistry and Western blot; meanwhile, the expressions of interleukin-1β（IL-1β） and interleukin-6（IL-6） m RNA were determined by real-time PCR. Data showed that there were pathological changes in Pp group and IP groups, but the injury was milder in IP groups than that in Pp group. The activity of SOD and T-AOC were significantly higher and MDA were significantly lower in all IP groups than those in Pp group（P〈0.05）; the expression of Bcl-2 protein elevated, and the expression levels of Bax protein and IL-1β, IL-6 m RNA decreased, especially in M-IP group. All results indicated that CO2 ischemic preconditioning has protective effects on liver against ischemia reperfusion injury in a rat model of CO2 pneumoperitoneum in pressure dependent manner, and 10 mm Hg intra-abdominal pressure may be more effective.The underlying m

关 键 词：缺血预处理 CO2气腹 肝脏 缺血/再灌注损伤 

分 类 号：R656[医药卫生—外科学] R-332[医药卫生—临床医学]