一贯煎影响CCl_4大鼠肝硬化形成期肝组织基因表达谱的效应机制研究

Hepatic Gene Expression Analysis of Carbon Tetrachloride-Induced Rat Cirrhosis Identifies Distinct Effects of Yiguanjian Decoction

目的:探讨一贯煎对CCl4诱导大鼠肝硬化形成阶段肝组织基因表达谱的影响。方法:首次大鼠皮下注射CCl43mL/kg,此后以50%CCl4橄榄油溶液2mL/kg皮下注射,每周2次,共计12周,自第9周开始给与药物干预4周,12周末杀鼠取材;检测肝功能,肝组织病理,肝组织羟脯氨酸含量,并采用基因芯片技术探讨一贯煎对模型大鼠肝组织基因表达谱的影响。结果:(1)模型大鼠8周时呈慢性肝损伤肝纤维化的病理改变,12周时已形成典型的肝硬化;与正常大鼠比较,12周时模型大鼠精氨酸加压素1A受体(AVPR1A)、CYP3A13、β球蛋白(Beta-glo)等基因表达显著下调;淋巴毒素A(LTA)、MMP-23、RNA结合基序蛋白3(RBM3)、血小板反应蛋白2(TSP2)、AP1γ亚单位结合蛋白1(AP1GBP1)、生长素释放素受体(GHRHR)、阿米洛利结合蛋白1(ABP1)等基因表达显著上调。(2)与12周模型对照组比较,一贯煎组AVPR1A、CYP3A13、Beta-glo等基因表达显著上调;LTA、MMP-23、RBM3、TSP2、AP1GBP1、GHRHR、ABP1等基因表达显著下调。结论:一贯煎抑制CCl4诱导大鼠肝硬化形成的作用机制与提高肝脏生物转化功能、抑制肝脏炎症反应、抑制肝细胞凋亡和肝星状细胞活化、抑制肝窦内皮损伤、改善肝脏糖代谢及水钠潴留等多种途径有关。

This paper reports an investigation into the effects of Yiguanjian decoction on hepatic gene expression in carbon tetrachloride - induced rat cirrhosis. In the investigation, pure carbon tetrachloride was injected into rats subcutaneously at a dose of 3ml· kg^-1 for the first time, and then 50% CC14 -olive solution at a dose of 2ml· kg^-1 twice a week for the following 12 weeks to induce cirrhosis. Yiguanjian decoction was administered orally starting from the 9th week for 4 weeks. Sera and liver tissues were collected at the end of 12th week to determine liver＇s functionalities, hepatic pathohistology, and hepatic hydroxyproline content. Meanwhile, oligonucleotides microarrays analysis was conducted to measure the effects of Yiguanjian decoction on hepatic gene expression. Histopathological examination showed that a chronic liver injury and liver fibrosis appeared in the 8th week, and typical cirrhosis formed in the 12th week for model group. Microarray analysis revealed that arginine vasopressin receptor 1A （AVPR1 A） , cytochrome P450 family 3, subfamily a, polypeptide 13 （CYP3A13） and Betaglo were down -regulated for model group in the 12th week, with another 7 genes up - regulated simultaneously, including lymphotoxin A （LTA） , matrix metalloproteinase 23 （ MMP - 23 ） , RNA binding motif protein 3 （ RBM3 ） , thrombospondin 2 （TSP2） , AP1 gamma subunit binding protein 1 （ AP1GBP1 ） , growth hormone releasing hormone receptor （GHRHR） , and amiloride binding protein 1 （ ABP1 ）. It is worth mentioning that genes down - regulated in model group up - regulated significantly in Yiguanjian decoction - treated group, along with those noticeably up - regulated or down - regulated. It is concluded that Yiguanjian decoction inhibits CCl4 - induced cirrhosis in rat significantly, and the underlying pathogenesis involves an enhanced hepatic biotransformation, inhibiting inflammation, lowering hepatocyte apoptosis and hepatic stellate cell activation, reducing hepatic sinusoidal endothelial cell injury, and improving hepatic glucoregulation and sodium and water retention.