低温机械灌注通过上调Kruppel样因子2减轻肝脏复灌后炎症反应  

Hypothermic Machine Perfusion（HMP） attenuates liver inflammation response after reperfusion through upregulating expression of KLF2

作　　者：胡晓燕[1] 曾承[1] 王伟[2] 王垒[1] 何维阳 方泽鸿 王彦峰[1] 叶啟发[1] 

机构地区：[1]武汉大学中南医院武汉大学肝胆疾病研究院武汉大学移植医学中心移植医学技术湖北省重点实验室,430071 [2]中南大学湘雅三医院卫生部移植医学工程技术研究中心

出　　处：《中华器官移植杂志》2016年第8期482-487,共6页Chinese Journal of Organ Transplantation

基　　金：国家自然科学基金新疆联合基金（U1403222）;中央高校基本科研业务基金（2042015kf1015）

摘　　要：目的观察低温机械灌注（HMP）中Kruppel样因子2（KLF2）与核因子κB（NF-κB）P65的表达及其对无心跳大鼠肝脏复灌后炎症反应的作用。方法采用随机数字表法将12只成年雄性SD大鼠分为HMP组（实验组），冷保存组（CS组，对照组），每组6只。剪开膈肌，待大鼠心脏停跳30min后获取肝脏，两组肝脏分别行HMP或CS3h，最后采用大鼠离体常温复灌系统模拟肝移植复灌的过程。复灌过程中，计算肝脏的灌注阻力指数，监测肝酶活性和氧耗量，记录胆汁生成量。复灌结束（2h）后，取两组肝组织样本进行病理学检查，采用免疫印迹法检测肝组织中KLF2、NF-κBP65的相对表达水平，采用酶联免疫吸附试验（ELISA）检测肝组织中肿瘤坏死因子α（TNF-α）、白细胞介素1β（IL-1β）的表达水平。结果复灌0、60和120min时，HMP组肝脏灌注液中丙氨酸转氨酶（ALT）、天冬氨酸转氨酶（AST）及乳酸脱氢酶（LDH）含量均显著低于CS组（P〈0．05），氧耗量均显著高于CS组（P〈0．05）；HMP组肝脏的胆汁生成量高于CS组，但两组差异无统计学意义（P〉0．05）。病理检查结果显示，HMP组大鼠肝脏的肝窦略扩张，有少量细胞出现水肿；CS组肝组织广泛水肿，胞浆疏松、淡染，肝窦淤血并伴有炎症细胞点状浸润。HMP组大鼠肝组织的KLF2相对表达量明显高于CS组（P〈0．05），CS组大鼠肝组织的NF-κBP65、TNF-α和IL-D的表达量明显高于HMP组（P〈0．05）。结论无心跳大鼠肝脏应用HMP保存可通过上调KLF2表达、抑制NF-κBP65的表达，从而减轻无心跳大鼠肝脏复灌后的炎症反应。Objective The aim of this research was to investigate the expression of KLF2 and nuclear factorkappa B P65 （NF-κB P65） in hypothermie machine perfusion and its role in liver inflammatory response after rat liver transplantation from non-heart-beating donor. Methods Twelve rats were randomly divided into two groups： HMP group（n = 6） and cold storage （CS） group（n = 6）. With 30min of hepatic pedicle occlusion, the rats were sacrificed and liver were obtained. HMP group experienced HMP and CS group experienced CS for 3h, respectively. Using the Isolated perfused rat liver model （IPRL） to mimick the reperfusion after transplantation. During normothermic reperfusion, intrahepatic resistance fIR） was calculated, the activities of hepatic enzyme in perfusate and oxygen consumption were monitored and production of bile was evaluated. After normothermic reperfusion, livers were obtained for histological study, the expression of Kruppel-like Factor 2 （KLF2） and NF-κBP65 were detected by Western Blot and the expression of TNF-α and IL-1β in liver were detected by ELISA. Results 1. After 0min, 60min, 120min of reperfusion, levels of ALT. AST. LDH in HMP group were significant lower than these in CS group （P〈0. 05）, while the oxygen consumption were significant higher in HMP group （P〈0. 05）, but the bile production had no difference with each other （P〉0. 05）. 2. In HMP group, hepatic sinusoids were somewhat dilated and only a very few hepatic cell edema, while CS group showed a serious liver injury including large hepatic cell edema, cytoplasmic loose, congestion and inflammation cells in the hepatic sinusoidal. 3. The expression of KLF2 in HMP group were significantly higher than CS group（P〈0. 05）, while the expression of NF- κBP65 .TNF-αand IL-1βin HMP group were significantly lower than CS group （P〈0. 05）. Conclusion HMP could decrease liver inflammation response after liver reperfusion of non-heart-beating donor through upregulating KLF2 and inhibiting NF-κ

关 键 词：大鼠 肝脏 低温机械灌注 冷保存Kruppel样因子2 

分 类 号：R575[医药卫生—消化系统]