大鼠酒精性肝病Kupffer细胞Toll样受体4和细胞因子的变化

The role of Toll-like receptor 4 in Kupffer cells and cytokines in alcohol-induced liver disease in rats

目的 观察酒精性肝病大鼠Kupffer细胞 (KCs)Toll样受体 (TLR) 4基因表达和细胞因子的变化。 方法 2 8只Wis tar大鼠随机分为乙醇喂养组 (E组 )和葡萄糖喂养组 (C组 )。E组大鼠饮水中加入乙醇 (剂量 5～ 12g.kg-1.d-1)喂养 ,C组饮水中加入等量的葡萄糖。两组大鼠分别于 4周和 8周活杀分离KCs,用逆转录多聚酶联反应 (RT PCR)测定KCs中TLR4mRNA的表达 ;用酶联免疫法 (ELISA)测定血浆中TNF α和IL 6浓度变化。结果 RT PCR显示E组TLR4mRNA的表达也显著高于C组(P <0 .0 1)。E组 4周和 8周的TNF α浓度分别为 (32 6± 4 2 )ng/L和 (40 2± 5 1)ng/L ,明显高于对照组的 (86± 12 )ng/L和 (97±13)ng/L (P <0 .0 1) ;IL 6浓度为 (387± 4 6 )ng/L和 (413± 5 1)ng/L ,也显著高于C组的 (73± 10 )ng/L和 (78± 11)ng/L (P <0 .0 1)。结论 乙醇能诱导大鼠肝脏KCs表达TLR4基因 ,TLR4和细胞因子在大鼠酒精性肝脏损害中可能起作用。

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 and Cytokines in the liver injury in the rats with alcohol induced liver disease(ALD).Methods Twenty and eight Wistar rats were divided into two groups: ethanol fed group (group E) and control group (group C). Group E were fed ethanol(dose of 5～12g.kg -1 .d -1 ) and group C received dextrose instead of ethanol. Rats of the two groups were sacrificed at 4 wk and 8 wk. KCs were isolated . TLR 4 mRNA in KCs was determined by the reverse transcription polymerase chain reaction (RT PCR) analysis. The levels of tumor necrosis factor (TNF) α and interleukin (IL) 6 in plasma were determined with enzyme linked immunsorbent assay (ELISA). Results Ethanol administration led to a significant increase in TLR 4 mRNA expression compared with group C (P<0.01). The concentrations of TNF αand IL 6 were (326±42 )ng/L, (402±51) ng/L, (387±46) ng/L and (413±51)ng/L, respectively,in group E which were higher than that in group C (86±12 )ng/L,(97±13)ng/L, (73±10 ) ng/L and(78±11) ng/L at 4 wk and 8 wk ( P <0.01). Conclusion Ethanol administration can markedly led to the gene expression of TLR 4 in KCs, and TLR 4 and cytokines may play a major role in the development of alcohol induced liver injury.