地塞米松在D-氨基半乳糖联合内毒素诱导小鼠急性肝功能衰竭  

Mechanisms of dexamethasone lreatment in acute hepatic failure induced by D-galactosamine combined with endotoxin in mice models

作　　者：甘建和[1] 吴旭东[1] 江敏华[1] 赵卫峰[1] 黄小平[1] 李国云[1] 

机构地区：[1]苏州大学附属第一医院感染病科,215006

出　　处：《中华传染病杂志》2007年第9期517-522,共6页Chinese Journal of Infectious Diseases

摘　　要：目的探讨地塞米松冲击疗法对D-氨基半乳糖(D-Gal)联合内毒素(LPS)所致急性肝功能衰竭模型促炎与抗炎因子的影响,为深入阐明糖皮质激素在重型肝炎中的作用机制提供实验依据。方法雄性BALB/c小鼠随机分为健康对照组6只、肝功能衰竭模型组24只和地塞米松治疗组21只,采用LPS 10μg/kg和D-Gal 900 mg/kg联合腹腔注射制备肝功能衰竭模型,治疗组同时注身地塞米松10mg/kg。健康对照组8 h,肝功能衰竭模型组和地塞米松治疗组2、4、6、8 h各6只小鼠心脏采血检测血清ALT、AST;肝组织经苏木精-伊红染色并应用病理学双盲评价行改良Knodell评分;不连续密度梯度法分离肝库普弗细胞(KCs)并鉴定,ELISA法检测KCs上清液IL-10、TNF-α和核因子(NF)-κB变化,半定量RT-PCR检测肝组织TNF-α、IL-10 mRNA表达。结果地塞米松治疗组与肝功能衰竭模型组组织学改良Knodell评分在6、8 h分别为7.500±1.871、10.830±1.172和10.500±1.049、15.670±1.633,差异有统计学意义(P<0.05);血清ALT、AST、肝KCs上清液TNF-α水平随损伤时间延长均逐渐升高,但治疗组较肝功能衰竭模型组低,差异有统计学意义(P<0.01),肝组织TNF-αmRNA亦随损伤时间增加而升高,治疗组与肝功能衰竭模型组在6、8 h比较,差异有统计学意义(P<0.05);肝KCs上清液及肝组织IL-10随损伤时间增加而升高,治疗组较肝功能衰竭模型组升高显著,在6、8h明显(P<0.05);肝KCs上清液NF-κB活性在2 h即升高,4 h达高峰,4～6 h为一高峰平台,8 h与正常无差异,两组2、4、6 h各时点之间差异有统计学意义(P<0.01)。结论肝功能衰竭早期应用地塞米松冲击疗法对肝脏有保护作用,其机制可能为通过抑制肝脏KCs NF-κB表达,下调肝脏原位及循环中TNF-α水平,调节促炎与抗炎因子之间的平衡而起作用。Objective To explore the effects of stosstherapy with dexamethasone on proinflammatory and anti-inflammatory factors in acute hepatic failure induced by D-galactosamine（D-Gal） and lipopolysaccharide in animal models and provide evidences for the mechanisms of glucocorticosteroid treatment in serious hepatic failure. Methods The male BALB/c mice were divided into three groups randomly： 6 in healthy control group, 24 in hepatic failure group and 24 in dexamethasone treated group. The mice were injected with lipopolysaccharide（LPS） 10 μg/kg and D-Gal 900 mg/kg intra- peritoneally to establish the hepatic failure model and dexamethasone was administrated simultaneously to the mice in treated group. The sera were collected from hearts of the mice 8 hours after injection in healthy control group and from 6 animals 2, 4, 6, 8 hours after injections in hepatic failure group and levels of alanine aminotransferase（ALT） and aspartate aminotransferase （AST） were tested. The reforming Knodell score was evaluated double-blindedly by hematoxylin and eosin staining and his- topathologic assay of liver tissue. Kuppffer cells（KCs） were isolated by discontinuous density gradient. Levels of interleukin（IL）-10, tumor necrosing factor（TNF）-α and nuclear factor kappa B（NF- kB） in the supernatant of KCs were detected by enzyme linked immunosorbent assay（ELISA）, IL-10 and TNF-α mRNA of liver tissue were determined by semi-quantitative reverse transcriptase polymerase chain reaction （RT-PCR）. Results Reforming Knodell scores 6, 8 hours after injections in trea- ted group and hepatic failure group were 7. 500±1. 871, 10. 830!1. 472 and 10. 500±1. 049, 15. 670 ± 1. 633, respectively, with statistically significant difference （P 〈 0.05）. Serum ALT, AST level and TNF-a level in KCs supernatant in these two groups increased gradually with prolonged liver injury and the level in treated group was lower than hepatic failure group （P 〈 0.01）. TNF-a mRNA in liver tissue also increased wit

关 键 词：肝功能衰竭 急性 地塞米松 NF—kB 小鼠 近交BALB/c 肿瘤坏死因子α 

分 类 号：R575.3[医药卫生—消化系统]