摘要
目的 :观察酒精性肝病大鼠Kupffer细胞 (KCs)Toll样受体 (TLR) 4蛋白合成和基因表达及其在肝损害中的作用。方法 :2 8只Wistar大鼠随机分为乙醇喂养组 (E组 )和葡萄糖喂养组 (C组 )。用激光共聚焦显微镜测定KCs膜上TLR4蛋白的合成 ;用逆转录多聚酶联反应 (RT -PCR)测定KCs中TLR4mRNA的表达 ;用鲎试剂基质偶氮显色法测定门静脉血中内毒素含量 ,并观察肝脏形态学变化。结果 :E组KCs膜TLR4蛋白表达较C组显著增强 ;E组TLR4mRNA的表达也显著高于C组 (P <0 .0 1) ;E组血浆内毒素浓度明显高于C组 (P <0 .0 1) ;E组肝组织发生显著的病理变化。结论 :乙醇能诱导大鼠肝脏KCs合成TLR4蛋白及表达TLR4基因 ,TLR4在乙醇所致的肝损害中可能起作用。
Objective:To observe the synthesis of Toll-like receptor (TLR)4 protein and its mRNA expression in Kupffer cells (KCs) and evaluate the role of TLR 4 in the liver injury in the rats with alcohol-induced liver disease (ALD).Methods:Twenty eight Wistar rats were divided into two groups: ethanol-fed group(group E) and control group (group C). Group E were fed with ethanol (dose of 5～12g/kg/d) and group C received dextrose instead of ethanol. Rats of the two groups were sacrificed at 4 wk and 8 wk. KCs were isolated and incubated.TLR 4 protein in KCs was determined by laser scanning confocal microscopy (LSCM).TLR 4 mRNA in KCs was determined by the reverse transcription polymerase chain reaction (RT-PCR).Plasma endotoxin levels were measured using the Limulus Amebocyte Lysate test kit.The liver pathology was observed under light and electronic microscopy.Results:The fluorescence intensity of TLR 4 protein in KCs by LSCM was stronger in group E than those in group C. Ethanol administration led to a significant increase in TLR 4 mRNA expression compared with group C ( P< 0.01).Plasma endotoxin levels increased significantly in group E ( 129±21)ng/L and (187±35 )ng/L at than in control rats (48±9)ng/L and (53±11)ng/L,respectively at 4 wk and 8 wk ( P <0.01). In liver section from group E,there was marked pathological changes including steatosis, cell infiltration and necrosis. None of the rats in group C developed marked pathological changes in liver. Conclusion: Ethanol administration can markedly lead to the synthesis of TLR 4 protein and its gene expression in KCs, and TLR 4 may play a role in the development of alcohol-induced liver injury.
出处
《重庆医科大学学报》
CAS
CSCD
2003年第4期444-447,共4页
Journal of Chongqing Medical University
基金
国家自然科学基金 (批准号 :3 0 170 919)
