大鼠急性肝衰竭模型自由基和细胞因子的动态变化

Dynamic changes of cytokines and free radicals in acute liver failure:experiment with rats

目的研究急性肝衰竭(ALF)时细胞因子(CK)和自由基(FR)的动态变化。方法64只SD大鼠接受D-氨基半乳糖(Galn)腹腔注射加脂多糖(LPS)静脉注射,以建立急性肝衰竭小鼠模型。LPS注射后0h,0.5h,1h,2h,4h,8h,16h,和24h,各处死8只大鼠,取肝组织和血清。以分光光度法检测血清中丙氨酸转氨酶(ALT)、天门冬氨酸转氨酶(AST)水平和肝匀浆中活性氧(AO)、丙二醛(MDA)、超氧化物歧化酶(SOD)含量;ELISA法检测血清和肝匀浆中肿瘤坏死因子α(TNF-α)和白细胞介素-1β(IL-1β)水平。结果血清ALT、AST活性在静注LPS后4h开始升高,16h达高峰;肝匀浆AO含量自0.5h开始即有上升,2h达峰值并维持在较高水平;肝匀浆MDA含量自LPS攻击后持续升高,SOD活性呈反相趋势。LPS攻击后,血浆和肝匀浆中的TNF-α水平在短时间内剧烈升高,2h达高峰,血清IL-11h达高峰,肝匀浆IL-1峰值在4h。结论CK和FR在ALF发生机制中起重要作用,并且早期就达到峰值。

Objective To investigate the dynamic changes of cytokines(CKs)and free radicals(FRs)in acute liver failure(ALF).Methods Sixty-four SD rats underwent intraperitoneal injection of D-galactosamine(D-GalN)and intravenous injection of lipopolysaccharide(LPS)to establish ALF models.Eight rats were killed at e different time points:0 h,0.5 h,1 h,2 h,4 h,8 h,16 h,and 24 h after the injection of LPS).Liver tissue and blood samples were collected.The serum levels of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)were measured by spectrophotometry.Spectrophotometry was used too to measure the levels of activated oxygen(AO),malondiadehyde(MDA),and superoxide dismutase(SOD)in the liver homogenate.The levels of tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)in serum and liver tissue were determined by ELISA.Results The serum ALT and AST levels began to increase 4h after the LPS injection,and peaked 16h after the injection.The AO level began to increase 0.5h after the LPS injection,peaked 2h later,and then stayed at a high level with a mild fluctuation.The MDA content of the liver homogenate increased continuously accompanied by decrease of the activity of SOD.The levels of TNF-αin the serum and liver homogenate increased dramatically 2h after the LPS injection.The levels of IL-1βin the serum and liver homogenate increased immediately after the injection of LPS and the peaked 1h and 4h after the injection respectively.Conclusion FRs and CKs play an important role in the mechanism of ALF.The production of FRs and CKs are increasing very rapidly soon after ALF occurs.