肝脏缺血再灌注损伤中线粒体膜氧化应激和电生理功能障碍的分子机制研究  

The pathogenesis of oxidative stress damage to mitochondrial membrane and electrophysiological dysfunction of ischemia reperfusion injury of hepatocytes in rats

作　　者：金涛[1] 王兴强[2] 刘超[3] 

机构地区：[1]天津市南开医院重症医学科,300100 [2]天津市第一中心医院移植ICU [3]天津市胸科医院心内科

出　　处：《天津医药》2016年第11期1347-1351,共5页Tianjin Medical Journal

基　　金：2013年中国博士后科学基金资助项目(2013M530880);2015年中国博士后科学基金资助项目(2015M581308)

摘　　要：目的探讨肝脏缺血再灌注损伤中线粒体膜氧化应激和电生理功能障碍的分子机制。方法建立SD大鼠肝移植(冷缺血再灌注组,n=20)和肝脏肝门部分阻断(热缺血再灌注组,n=20)的缺血再灌注损伤模型。以大鼠仅作肝十二指肠韧带分离,但不阻断肝脏肝门血液供应的10只大鼠为假手术组,采集各组血标本测定丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、肿瘤坏死因子-α(TNF-α)及白细胞介素-1β(IL-1β)水平;采集胆汁标本用于测定胆汁内葡萄糖的含量及γ-谷氨酰转移酶(GGT)水平;采集肝组织标本用于测定丙二醛(MDA)及超氧化物歧化酶(SOD),流式细胞仪测定肝细胞线粒体膜电位,采用酶联免疫吸附试验(ELISA)检测肝细胞内线粒体呼吸链复合体活性。结果冷、热缺血再灌注组大鼠的ALT、AST、胆汁葡萄糖、GGT、TNF-α、IL-1β及MDA水平均高于假手术组(P<0.01),且冷缺血再灌注组大鼠各项指标均高于热缺血再灌注组(P<0.05)。热缺血再灌注组和冷缺血再灌注组大鼠肝脏SOD水平均明显低于假手术组,并且热缺血再灌注组的SOD水平高于冷缺血再灌注组(P<0.01)。与假手术组相比,热、冷缺血再灌注损伤组再灌注0 h(缺血1 h)、1 h、12 h线粒体膜电位细胞的比例均降低(P<0.01);热、冷缺血再灌注损伤组组内随时间延长其线粒体膜电位活性有逐渐恢复的趋势(P<0.01)。假手术组0、72 h的线粒体呼吸链复合体Ⅲ、Ⅳ差异均无统计学意义;热、冷缺血再灌注组72 h较0 h线粒体呼吸链复合体活性均增高(P<0.01);0、72 h时,假手术组,热、冷缺血再灌注损伤组线粒体呼吸链复合体Ⅲ呈依次降低,而线粒体呼吸链复合体Ⅳ呈依次增高(P<0.01)。结论缺血再灌注损伤的强应激刺激可导致肝细胞线粒体膜氧化应激,进而导致肝细胞受到损害,线粒体内相关酶系统活性受损,最终导致线粒体内呼吸链酶复合体活性受损伤,这可能是氧化�Objective To explore the pathogenesis of oxidative stress damage to mitochondrial membrane andelectrophysiological dysfunction of ischemia reperfusion injury of hepatocytes in rats. Methods Seventy rats were randomlydivided into three experimental groups: sham operation(SHAM) group, warm hepatic ischemia/reperfusion(w I/R) group andcold hepatic ischemia/reperfusion(c I/R) group. Blood samples were collected for the detection of alanine aminotransferase(ALT), aspartate aminotransferase(AST), tumor necrosis factor-α(TNF-α) and interleukin(IL)-1 β levels. Bile sampleswere collected for the detection of glucose and γ-glutamyl transferase(GGT) levels. And liver samples were collected for thedetection of malondialdehyde(MDA) and superoxide dismutase(SOD). Flow cytometry was used to detect mitochondrialmembrane potential. The mitochondrial respiratory chain complex was examined using ELISA to assess ischemia reperfusioninjury of hepatocytes in rats. Results The results indicated that ALT, AST, GLU, GGT, TNF-α, IL-1β and MDA levelswere increased significantly in the w I/R group and c I/R group than those in SHAM group(P<0.01), and those indexes weresignificantly higher in c I/R group than those of w I/R group(P<0.05). The SOD level was significantly lower in w I/R groupand c I/R group than that in SHAM group, which was significantly higher in w I/R group than that of c I/R group(P<0.01).Compared with SHAM group, ratios of mitochondrial membrane potential were significantly decreased at 0, 1 and 12 h I/R in w I/R group and c I/R group(P<0.01). The activity of mitochondrial membrane potential was gradually recovered with time in w I/R group and c I/R group(P<0.01). There were no significant differences in mitochondrial respiratory chain complexes Ⅲand Ⅳ between 0 h and 72 h in SHAM group. The activity of mitochondrial respiratory chain complexes was increased at 72 h than that of 0 h in w I/R group and c I/R group(P<0.01). The activity of mitochondrial respiratory chain complexes Ⅲ was decreased ordinally at 0

关 键 词：肝移植 再灌注损伤 疾病模型 动物 氧化应激 线粒体跨膜电位 肝脏损伤 

分 类 号：R657.3[医药卫生—外科学] R364.12[医药卫生—临床医学]