构建启动大鼠肝组织中TGF-β/Smad信号传导通路的急性肝损伤动物模型

目的在传统CCl4急性肝损伤模型的基础上,构建启动大鼠肝组织中的TGF-β/Smad信号传导通路的急性肝损伤动物模型。方法将30只大鼠随机分为3组,每组各10只,分别为模型组,对照组和空白组,模型组大鼠予小动脉夹夹闭肝总动脉15min手术处理,随后分别在第6天和第10天,予25%CCl4花生油溶液6mL/kg体重腹腔注射;对照组则单用两次CCl4,空白组不做任何处理;第二次CCl448h后处死所有动物。结果模型组与对照组及空白组比较：血清指标ALT、AST、HA显著上升（P〈0.01）;肝组织HE染色病理观测,肝组织出现明显炎症、变性、坏死,纤维组织增生等现象;PT-PCR检测Ⅰ、Ⅲ型胶原、TGF-β1、Smad3mRNA表达显著增强（P〈0.01）;免疫组化检测Ⅰ、Ⅲ型胶原、TGF-β1、Smad3蛋白的表达增强（P〈0.01）。结论成功启动急性肝损伤大鼠肝组织中的TGF-β/Smad信号传导路,该急性肝损伤动物模型兼备肝纤维化活跃,和TGF-β/Smad信号传导通路信号增强特征,在评价早期抗肝纤维化药物及方法时具有耗时少、有效和经济的特点,值得进一步研究。

Construction of an Acute Liver Damage Animal Model for Priming TGF-β/Smad Signal Conduction Pathway in Hepatic Tissue of Rats

Objective To construct an acute liver damage animal model for priming TGF-β/Smad signal conduction pathway in hepatic tissue of rats on the basis of traditional model induced by carbon tetrachloride （ CCl4 ）. Methods Thirty rats were randomly divided into three groups as model group,control group and blank group. The model group was prepared by clamping the hepatic artery communis with bulldog clamp for 15 minutes was injected intraperitoneally with 25% of CCl4 contained in arachis oil on the following sixth and tenth day after the operation,respectively. The dosage of the liquor was 6 mL per kg. The control group was only injected intraperitoneally with CCl4 twice,while the blank group without any treatment. After 48 hours of the second injection,the rats were sacrificed. Results The levels of ALT,AST and HA in plasma were apparently increased in model group compared with that in control group and blank group （ P 0. 01 ）. The liver HE staining in model group showed that there were obvious inflammation,apomorphosis,cellular necrosis and fibroplasias,the gene expression and protein expression of type Ⅰ,Ⅲ collagen,TGF-β1 and Smad3 enhanced obviously with RT-PCR detection and Immunohistochemistry technology in the model group than that in control group and blank group （ P 0. 01）. Conclusion Animal model of acute liver injury have two features：active liver fibrosis and TGF-β /Smad signaling pathway signal enhancement. The acute liver injury rat model priming TGF-β /Smad signal conduction pathway in liver tissue has advantages of less time consuming,utility and economy in evaluating the drugs and methods for early anti-hepatic fibrosis.