二硫代氨基甲酸吡咯烷对小鼠急性凋亡性肝损伤的效应  

Effects of pyrrolidine dithiocarbamate on acute apoptotic liver injury in mice

作　　者：王华[1] 吕金伟[1] 张程[1] 宁萑[1] 赵磊[1] 徐德祥[1] 

机构地区：[1]安徽医科大学卫生毒理学系,安徽合肥230032

出　　处：《中国药理学通报》2009年第12期1610-1614,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No30572223);安徽医科大学科研基金项目;追加基金资助项目(No2006kj13和2008kjzj01)

摘　　要：目的探讨二硫代氨基甲酸吡咯烷(PDTC)对氨基半乳糖(GalN)/细菌脂多糖(LPS)诱导小鼠急性凋亡性肝损伤的作用及其分子机制。方法设置生理盐水(NS)组、GalN/LPS组、PDTC+GalN/LPS组和PDTC组。每组10只小鼠被用于观察LPS处理后72h内的动物死亡情况;每组6只小鼠经LPS处理后1.5h被取血、处死并留取肝脏,用RT-PCR检测肝脏组织TNF-αmRNA表达水平,用EMSA分析肝脏NF-κB结合活性,每组12只小鼠于LPS处理后8h取血、处死并留取肝脏,测定血清丙氨酸转氨酶(ALT)活力,用TUNEL技术检测肝脏细胞凋亡,并对肝组织切片行常规HE染色。结果GalN/LPS共处理升高小鼠血清ALT活力;肝脏组织病理学检查发现,GalN/LPS组小鼠肝脏严重充血、坏死并伴有大量炎性细胞浸润,肝脏组织TUNEL阳性细胞增多;在GalN/LPS共处理72h内有90%小鼠发生死亡,所有死亡小鼠均伴有肝脏严重充血。PDTC预处理抑制GalN/LPS诱导的肝脏NF-κB激活和TNF-α表达,但PDTC预处理反而加重GalN/LPS引起的小鼠肝脏细胞凋亡、进一步升高血清ALT活力、加重肝脏充血和坏死并加速小鼠死亡。结论NF-κB抑制剂PDTC通过抑制肝脏实质细胞NF-κB介导的抗凋亡机制加重GalN/LPS诱导的小鼠急性凋亡性肝损伤。Aim To investigate the effects of pyrrolidine dithiocarbamate （PDTC） on D-galactosamine/lipopolysaccharides （GalN/LPS）-induced acute apoptotic liver injury and its mechanism.Methods All mice were randomly divided into four groups.Mice in GalN/LPS group were co-injected with GalN （600 mg·kg^-1ip） and LPS （20 μg·kg^-1,ip）.Mice in PDTC＋GalN/LPS group were injected with two doses of PDTC,one （100 mg·kg^-1ip） at 24 h before LPS and the other at 2 h before LPS （20 μg·kg^-1,ip）.Mice in control groups were treated with PDTC （100 mg·kg^-1ip） or saline.Ten mice in each group were observed for animal survival within 72 h after LPS treatment.Six mice in each group were sacrificed 1.5 h after LPS for collecting blood and isolating livers.The expression of hepatic TNF-α mRNA was determined by reverse transcription and polymerase chain reaction （RT-PCR）.Hepatic nuclear factor-κB （NF-κB） binding activity was measured with electrophoretic mobility shift assay （EMSA）.Twelve mice in each group were sacrificed 8 h after LPS treatment.Serum was collected for measurement of alanine aminotransferase （ALT） and hepatocellular apoptosis and histological examination.Results Co-injection of GalN and LPS markedly increased serum ALT activity.Histopathological examination of liver sections revealed that GalN/LPS induced hepatic congestion,necrosis and massive macrophages infiltration,and increased the number of TUNEL-positive cells in mouse liver.GalN/LPS treatments,led to 90% mortality within 72 h with severe congestion and necrosis in the liver of all the dead mice.PDTC pretreatment significantly inhibited GalN/LPS-induced hepatic NF-κB activation and TNF-α expression.In contrast,PDTC aggravated GalN/LPS-triggered hepatocellular apoptosis,increased serum ALT activity,exacerbated hepatic hemorrhage and necrosis,and accelerated death.Conclusion PDTC aggravates GalN/LPS-induced acute apoptotic liver injury via inhibiting NF-κB-mediated anti-apoptotic effects.

关 键 词：二硫代氨基甲酸吡咯烷 脂多糖 氨基半乳糖 急性凋亡性肝损伤 核因子-ΚB 小鼠 

分 类 号：R-332[医药卫生] R322.47