实验性急性肝衰竭大鼠血清中TNF-α的来源及水平变化

Source and Level Change of Sera TNF-α in Rats of Experimental Acute Hepatic Failure

目的研究D-氨基半乳糖/内毒素所致大鼠急性肝衰竭模型中血清内TNF-α的来源及不同时间点的水平变化。方法取30只Wistar大鼠腹腔内注射D-氨基半乳糖/内毒素诱导大鼠急性肝衰竭模型(AHF组,n=30),于建模3 h、6 h1、2 h2、4 h4、8 h7、2 h各取5只动物检测其血清丙氨酸转氨酶(ALT)与肝组织的病理形态学变化;采用ELISA检测不同时间点动物血清中TNF-α水平的变化;通过免疫组化法检测不同时间点动物肝、肺组织内TNF-α的表达。另取30只Wistar大鼠腹腔内注射等量生理盐水为正常对照(N组,n=30)。结果AHF组各时间点血清ALT的增高与N组相比有统计学差异(P<0.05),肝脏内炎细胞浸润、坏死明显;AHF组与N组相比血清内TNF-α的增高在3 h(P<0.01)6、h(P<0.05)有统计学差异;AHF组肺组织与N组相比TNF-α的表达在3 h、6 h、12h(均为P<0.01)有统计学差异。结论本实验成功建立了大鼠AHF模型;TNF-α在此模型的早期阶段起重要作用;肺脏可能是血清内TNF-α的主要来源之一。

Objective To study the source and level change of sera TNF-α in D-glactosamine/endotoxin-induced acute hepatic failure models. Methods Wistar rats with acute hepatic failure （AHF group, n = 30） were induced by intrapefitoneal injection of D-glactosamine/endotoxin. To evaluate necrosis of hepatic ceils, sera Alanine Transaminase （ALT） were determined and liver pathological morphology were observed at 3 h,6 h,12 h,24 h,48 h and 72 h following intraperitoneal injection of D-glaetosamine and endotoxin, sera TNF-α were examined by ELISA; the expression of TNF-α in livers and lungs were detected by immunohistochemistry technique. Wistar rats were injected with physiological saline solution as normal controls （N group, n = 30）. Results Sera ALT increased significantly in AHF group than in N group （P 〈 0.05 ） at all time points, infiltration of inflammatory ceils and necrosis of hepatic ceils were marked in AHF group; Compared with N group, levels of sera TNF-α in AHF group were obviously higher at 3 h（ P 〈 0.01 ） and 6 h（ P 〈 0.05） ; the expression of TNF-α in lungs in AHF group were markedly stronger than in N group at 3 h ,6 h and 12 h（ P 〈 0.01 ,for these time points）. Conclusion AHF rat＇ s models were successfully founded in our experiment; TNF-α may play important roles in the AHF models at early phase;Lungs may be the major source of sera TNF-α.