自制肝保护液对梗阻性黄疸大鼠门静脉阻断所致肝细胞损伤的保护作用  

Protective effect of self-made liver solution on hepatocyte injury in obstructive jaundice rats after portal vein occlusion

作　　者：冯春林[1,2] 林恒[1] 陈炜[2] 冷凯[2] 郑晓华[2] 主鹤亭[2] 别平[1] 

机构地区：[1]第三军医大学西南医院全军肝胆外科研究所,重庆400038 [2]遵义医学院第三附属医院肝胆外科,贵州遵义563000

出　　处：《第三军医大学学报》2011年第15期1563-1567,共5页Journal of Third Military Medical University

摘　　要：目的探讨自制肝保护液对梗阻性黄疸大鼠门静脉阻断所致肝细胞损伤的作用及对肝细胞MAPKs信号途径的影响。方法 72只Wistar系大鼠完全随机分为4组:A组:胆道再通、门静脉周围游离、切除肝尾状叶,90 min后关腹腔;B组:胆道再通、肝尾状叶分流门静脉、余肝门静脉血流阻断90 min、恢复正常门静脉血流、切除肝尾状叶,关腹;C组:B组+门静脉肝侧灌注4℃乳酸林格氏液;D组:B组+门静脉肝侧灌注4℃自制肝保护液(home-made liver solution,HMLS)。术后0、6、24 h获取各组大鼠组织标本。采用RT-PCR和Western blot分别测定肝组织A20 mRNA和蛋白表达,测定肝组织中p-JNK、p-ERK、p-p38、Bcl-2、Bax和Caspase-3的蛋白表达情况,TUNEL法检测肝细胞凋亡指数。结果 D组大鼠A20 mRNA和蛋白表达均明显增加(P<0.05),p-JNK、Bax、Caspase-3蛋白表达明显下降(P<0.05),而p-ERK、Bcl-2蛋白表达明显增加(P<0.05)。C、D组大鼠与B组相比肝细胞凋亡率明显下降(P<0.05),其中D组下降更明显。结论自制肝保护液经门静脉在体持续低温灌注肝脏,可以促进A20 mRNA及其蛋白的高表达,由此激活ERK信号转导通路,抑制JNK信号转导通路,反向调节线粒体途径Bcl-2/Bax蛋白平衡的逆转,降低了其下游的凋亡执行蛋白Caspase-3,从而起到抗凋亡、保护肝细胞的作用。Objective To explore the effect of self-made liver solution on hepatocyte injury and MRPK signal transcriptional pathway after portal vein occlusion in rats with obstructive jaundice. Methods Seventy-two Wistar rats were randomly divided into four groups, group A （ biliary tract recanalization, liberation of portal vein, resection of caudate lobe of liver and closure of abdomen in 90 rain） , group B （ biliary tract recanalization, portal vein bypass in caudate lobe of liver, PVO of left hepatic tissue in 90 min, then recovery of portal vein flow and resection of caudate lobe of liver）, group C （ group B with perfusion of 4 ℃ Lactated Ringer＇s Solution） and group D （group B with portal vein perfusion of 4 ℃ self-made liver solution）. Liver samples were taken at 0, 6 and 24 h post operation. Reverse transcription PCR, Western blotting and TUNEL were used respectively to determine A20 mRNA, A20 protein, p-JNK, p-ERK, p-p38, Bcl-2, Bax, Caspase-3 and hepatocyte apoptotic rate. Results The levels of A20 mRNA and protein were increased obviously in group D compared to the other groups （ P 〈 0.05 ）. The expression of p-JNK, Bax, and Caspase-3 was decreased but the expression of p-ERK and Bcl-2 protein was presented the adverse changes. The hepatocyte apoptotic rates of group C and group D were obvious lower than that of group B. Conclusion Continuous hypothermical perfusion with self-made liver solution improves expression of anti-apoptosis protein A20, then activates ERK signal transduction pathway to regulate the balance of Bcl-2/Bax protein and decreases the expression of apoptosis executive protein Caspase-3.

关 键 词：梗阻性黄疸 门静脉血流阻断 自制肝保护液 MAPK信号转录途径 

分 类 号：R318.52[医药卫生—生物医学工程] R322.47[医药卫生—基础医学]