法舒地尔对梗阻性黄疸大鼠肝缺血肝切除后引起急性肝衰竭的作用机制研究

Mechanism of fasudil alleviating postoperative acute hepatic failure induced by hepatic is- chemia ＆ hepatectomy in rats with obstructive jaundice

目的：建立一种能较大程度模拟临床过程中对在梗阻性黄疸基础上进行肝缺血肝切除后引起急性肝衰竭的动物模型,并进一步探讨法舒地尔对本实验模型的影响。方法：选取96只Wistar大鼠结扎切断胆总管制作梗阻性黄疸动物模型。急性肝衰竭大鼠随机分为对照组、低剂量法舒地尔治疗组（低剂量组）、高剂量法舒地尔治疗组（高剂量组）,每组各32只。低剂量组大鼠行肝切除后立即经门静脉注射法舒地尔10mg/kg,高剂量组门静脉注射法舒地尔30mg/kg,对照组门静脉注射等量生理盐水。检测肝衰竭大鼠术后6h血清谷丙转氨酶（ALT）、谷草转氨酶（AST）、总胆红素（TBIL）、肿瘤坏死因子-α（TNF-α）和干扰素-γ（INF-γ）水平;检测肝组织中超氧化物歧化酶（SOD）活性及丙二醛（MDA）含量;肝组织HE染色观察病理损伤;观察模型动物96h生存率。结果：大鼠梗阻14d后行胆肠内引流,并阻断70%肝脏血供30min后切除余肝符合肝衰竭动物的标准,适合进一步研究。与对照组相比法舒地尔处理组中血清AST、ALT、TBIL、TNF-α、INF-γ水平下降,肝组织SOD活性增高,MDA含量降低,肝组织病理损伤减轻,大鼠96h生存率提高,且高剂量组较低剂量组效果更明显。结论：建立了一种发病机制类似于临床的外科型大鼠肝衰竭动物模型。另外,发现了法舒地尔可能通过调节炎症反应、降低肝脏脂质过氧化、加强自由基的清除,而对肝衰竭有保护作用。

Objective： To establish animal models of postoperative acute hepatic failure induced by hepatic ischemia No hepatectomy in rats with obstructive jaundice and to explore the mechanism how fasudil alleviat postoperative acute hepatic fail- ure. Methods： Selected 96 Wistar big rats as animal model of obstructive jaundice, which were treated with ligation and cuting off common bile duct. These rats with acute hepatic failure were randomly divided into control group, low dosage of fasudil treatment group （low-dose group）, high dosage of fasudil treatment group （high-dose group） with 32 rats in each group. Rats in low-dose group were injected with fasudil of 10mg/kg through portal vein immediately after hepatectomy, while rats in high- dose group were injected with fasudil of 30mg/kg through portal vein immediately after hepatectomy, rats in control group were injected with equivalent normal saline through portal vein immediately after hepatectomy. Serum ALT, AST, TBIL, （tumor necrosis factor-a, TNF-a）and （interferon-y, INF-7） levels in postoperative rats with hepatic failure were determined within 6 hours superoxide dismutase （SOD） activity and malondialdehyde （MDA） content in hepatic tissue were detected;pathological injury in hepatic tissues were detected with HE staining. Results： Rat models were successfully established. Com- paring with the control group, serum AST. ALT. TBIL. TNF-~. INF-~＇ levels of the observation group were significantly de- creased, SOD activities increased but MDA contents decreased. Pathological injury of the observation group were lighter and 96 h survival rate increased. The above mentioned difference was more obvious in the high-dose group than that in the low dose group. Oonclusion; A surgical hepatic failure model in rat is established, which shows similar clinical pathophysiological chan- ges as human. In addition, it was shown that fasudil possibly protect hepatic failure through regulating the inflammatory re- sponse, reducing hepatic lipid peroxidation and strengthening removal of the free radicals.