急性胆道梗阻大鼠肝细胞的死亡方式

Patterns of liver cell death in rats with acute bile duct obstruction

目的:通过建立大鼠急性胆道梗阻模型研究梗阻性黄疸时肝细胞死亡方式.方法:♂Wistar大鼠随机分为胆道结扎(BDL)组和对照(C)组.BDL组采用Yoshidome法建立急性胆道梗阻模型,7d后取材,应用流式细胞仪检测肝细胞胀亡和凋亡的百分比,HE染色和电镜观察肝组织病理组织学改变以及肝细胞凋亡、胀亡,并检测血清总胆红素TBIL,DBIL,ALP,γ-GT,ALT,AST和LDH水平.结果:BDL组胀亡和坏死细胞百分比均显著高于C组(胀亡:25.41%±2.18% vs 6.12%±1.69%;坏死:3.99%±1.24% vs 0.79%±0.29%;均P<0.05),而凋亡百分比显著低于C组(2.79%±1.43% vs 5.18%±1.87%,P<0.05).BDL组胀亡细胞百分比显著高于凋亡(25.41%±2.18% vs 2.79%±1.43%,P<0.05).肝病理组织学检查发现BDL组胆管增生,纤维结缔组织明显增生,肝细胞索排列紊乱,胀亡增加.BDL组血清TBIL,DBIL,ALP,γ-GT,AST,ALT和LDH水平均显著高于C组(P<0.05).结论:急性胆道梗阻时大鼠肝细胞死亡以胀亡为主,其肝功能损伤程度与肝细胞胀亡、凋亡有密切关系.

AIM： To investigate the patterns of hepatic cell death in rats with acute bile duct obstruction. METHODS： Male Wistar rats were divided randomly into bile duct ligation （BDL） group （n = 5） and control group （n = 5）. The rats in BDL group underwent ligation and division of common bile duct to set up acute bile duct obstruction model according to Yoshidome＇s maneuver, while the rats in the control group underwent dissociation of common bile duct only. All the animals were sacrificed for sampling on day 7. Flow cytometry was used to determine the percentages of hepatocyte apoptosis and oncosis. Hematoxylin and eosin （H＆E） staining and electron microscopy were used to observe hepatocyte apoptosis, oncosis and histopathological changes. An auto analyzer was used to examine the levels of serum total bilirubin （TBIL）, direct bilirubin （DBIL）, alkaline phosphatase （ALP）, glutamyl transpeptidase （γ-GT）, alanine aminotransferase （ALT）, aspartate aminotransferase （AST） and lactate dehydrogenase （LDH）. RESULTS： The percentage of liver cell oncosis or necrosis was significantly higher in BDL group than that in the control group （oncosis： 25.41%±2.18% vs 6.12%± 1.69%, P 〈 0.05; necrosis： 3.99% ± 1.24% vs 0.79% ± 0.29%, P 〈 0.05）, while the percentage of apoptosis was remarkably lower in BDL group （2.79% ± 1.43% vs 5.18% ± 1.87%, P 〈 0.05）. Moreover, the percentage of hepatic cell oncosis was higher than that of apoptosis in BDL group （25.41% ± 2.18% vs 2.79% ± 1.43%, P 〈 0.05）. Hepatic histopathological examination showed proliferation of bile duct and fibrous connective tissue, obvious increase of hepatic cell oncosis and liver cell cord derangement in BDL group. The levels of serum TBIL, DBIL, ALP, γ-GT, AST, ALT and LDH in BDL group were all increased as compared with those in the control group （P 〈 0.05）. CONCLUSION： Oncosis is the dominant type of hepatic ceU death in rats with acute bile duct obstruction, and the degree of hepatic injury has a close relationship with the oncosis or apoptosis of hepatic ceils.