TNF-α体外介导小鼠肝细胞凋亡和坏死  

TNF-α induced apoptosis and necrosis of mice hepatocytes

作　　者：臧国庆 俞红[1] 周霞秋[1] 廖丹[1] 谢青[1] 王斌[1] 

机构地区：[1]上海第二医科大学附属瑞金医院传染病科,上海市200025

出　　处：《世界华人消化杂志》2000年第3期303-306,共4页World Chinese Journal of Digestology

摘　　要：目的 探讨TNF-α在体外作用于小鼠肝细胞时肝细胞所发生的病理及生化改变。 方法 采用原位灌洗法分离小鼠肝细胞,体外培养后分别加入TNF-α,GalN和CalN/TNF-α 5,10,20和30h后,观察其形态学改变,测定培养上清液ALT和AST,抽提肝组织DNA,琼脂糖凝胶电泳观察生化学性质改变。 结果 TNF-α单独作用于培养肝细胞,无肝细胞凋亡和坏死,经GalN致敏后,TNF-α可诱导大量肝细胞凋亡,早期(10h)为凋亡,随后(20h以后)出现肝细胞坏死,其程度随培养时间延长和药物剂量的增加而明显,单独使用GalN也可造成肝细胞凋亡和坏死,但程度较GalN/TNF-α合用轻。 结论 TNF-α可诱导肝细胞凋亡和坏死,但须有肝细胞转录过程的抑制。AIM To investigate the effects of tumor necrosis factor-α (TNF-α) on hepatocytes.METHODS The hepatocytes were isolated in the way of liver perfusion in situ with 0.5g/L collagenase type Ⅳ and cultured with tumor necrosis factor-α (TNF-α) or D-galactosamine (GaIN) or TNF-α plus GaIN respectively for 5, 10, 20 and 30 h. Hematoxylin and esoin (HE) and Hoechst 33258 staining were used to observe the morphological changes of hepatocyte. DNA fragment was analyzed by the agrose gel electrophoresis to determine the hepatocyte apoptosis. The ALT (alanine aminotransferase) and AST (aspartate aminotransferase) in the medium supernatant were assayed for assessing necrosis.RESULTS Hepatocytes incubated with TNF-α plus GaIN showed that the number of hepatocyte apoptosis increased in parallel with the dose of TNF-α and incubation time. The hepatocyte apoptosis was further confirmed by DNA ladder. After 20 hours, hepatocyte necrosis took place with the increase of ALT and AST. In addition, hepatocyte incubated with GaIN also caused small amounts of hepatocyte apoptosis and necrosis. CONCLUSION TNF-α can induce hepatocyte apoptosis in the presence of transcription inhibition of hepatocyte.

关 键 词：肝细胞 肿瘤坏死因子 细胞凋亡 肝坏死 

分 类 号：R361[医药卫生—病理学]