CAJ | 학술논문

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72 h内的动物死亡情况;每组6只小鼠经LPS处理后1.5 h被取血、处死并留取肝脏,用RT-PCR检测肝脏组织TNF-α mRNA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8 h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72 h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.
목적 탐토이류대안기갑산필각완(PDTC)대안기반유당(GalN)/세균지다당(LPS)유도소서급성조망성간손상적작용급기분자궤제.방법 설치생리염수(NS)조、GalN/LPS조、PDTC+GalN/LPS조화PDTC조.매조10지소서피용우관찰LPS처리후72 h내적동물사망정황;매조6지소서경LPS처리후1.5 h피취혈、처사병류취간장,용RT-PCR검측간장조직TNF-α mRNA표체수평,용EMSA분석간장NF-κB결합활성,매조12지소서우LPS처리후8 h취혈、처사병류취간장,측정혈청병안산전안매(ALT)활력,용TUNEL기술검측간장세포조망,병대간조직절편행상규HE염색.결과 GalN/LPS공처리승고소서혈청ALT활력;간장조직병이학검사발현,GalN/LPS조소서간장엄중충혈、배사병반유대량염성세포침윤,간장조직TUNEL양성세포증다;재GalN/LPS공처리72 h내유90%소서발생사망,소유사망소서균반유간장엄중충혈.PDTC예처리억제GalN/LPS유도적간장NF-κB격활화TNF-α표체,단PDTC예처리반이가중GalN/LPS인기적소서간장세포조망、진일보승고혈청ALT활력、가중간장충혈화배사병가속소서사망.결론 NF-κB억제제PDTC통과억제간장실질세포NF-κB개도적항조망궤제가중GalN/LPS유도적소서급성조망성간손상.
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~(-1),ip) and LPS (20 μg·kg~(-1), ip). Mice in PDTC+GalN/LPS group were injected with two doses of PDTC,one (100 mg·kg~(-1), ip) 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~(-1), ip) 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.