慢加急性肝衰竭大鼠血清TNF-α、白介素-10及肝细胞凋亡的动态变化

Kinetic changes of serum TNF-a,IL-10 and hepatocytes apoptosis in rat’s acute on chronic liver failure model

目的建立大鼠慢加急性肝衰竭实验模型,观察其血清TNF-α、IL-10及肝细胞凋亡的动态变化。方法首先用人血白蛋白建立大鼠肝硬化模型,至肝纤维化4级时予D-氨基半乳糖/脂多糖急性攻击,计算动物死亡率及生存时间,观察给药后4h、8h、12h肝功能、血清TNF-α、IL-10水平及病理变化,TUNEL法检测原位细胞凋亡,计算凋亡指数。结果D-氨基半乳糖/脂多糖攻击后90%大鼠死于急性肝衰竭,平均生存时间(16.1±3.7)h,转氨酶及胆红素的变化符合肝功能衰竭特点,病理表现为肝硬化再生结节内发生大块或亚大块坏死,纤维间隔保留。血清TNF-α明显增高并与肝细胞凋亡程度相一致。IL-10随给药时间延长而增高,与临床慢加急性肝衰竭患者变化相似。结论对人血白蛋白免疫诱导型肝硬化大鼠给予D-氨基半乳糖/脂多糖联合急性攻击可建立慢加急性肝衰竭模型。TNF-α介导的肝细胞凋亡可能是该慢加急性肝衰竭重要病理机制之一,提示早期给予人工肝及血液净化治疗清除炎性介质、阻断炎症反应可能对慢加急性肝衰竭起到一定防治作用。

Objective To establish experimental model of acute on chronic liver failure in rats, describing the kinetic changes of serum TNF-α,IL-10 and hepatoeytes apoptosis. Methods Immunological hepatic fibrosis was induced by human serum albumin injection in Wistar rats. In rats with fibrosis stage Ⅳ, D-galaetosamine/ lipopolysaeeharide were administered. Mortality and survival time were recorded in 20 rats. Ten rats were sacrificed at 4, 8, and 12 hours, respectively. Liver function parameters, serum TNF-α,IL-10 levels were measured after treatment with D-galaetosamine/lipopolysaceharide, as well as liver pathology study. Cell apoptosis was detected by tunnel assay. Results 90% rats died from acute liver failure after administration of D-galaetosamine / lipopolysaeeharide, with mean survival time of （16.1 ± 3.7） hours.Liver function tests were compatible with liver failure. Histopathology revealed massive or sunbmassive necrosis in regenerative nodules, while fibrosis septa were intact. Plasma level of TNF-α significantly increased, in association with apoptosis, while profile of plasma IL-10 level was consistent with immunosuppression seen in patients with acute-on-chronic liver failure. Conclusion Experimental model of acute-on-chronic liver failure can be established successfully by administration of D-galaetosamine/lipopolysaeeharide on the immunological-induced cirrhosis. TNF-α-mediated hepatoeytes apoptosis may play very important role in the pathogenesis. This results indicated that early artificial liver or blood apheresis therapy which could remove inflammatory medium may be useful.