血府逐瘀汤联合茵栀黄对肝纤维化大鼠的保护作用研究

The Protective Effect of Xuefuzhuyu Decoction combined with Yinzhihuang on Hepatic Fibrosis in Rats

目的:研究血府逐瘀汤联合茵栀黄对二甲基亚硝胺(DMN)致肝纤维化大鼠的保护作用。方法:将68只雄性SD大鼠按体重随机分为6组:正常对照组、秋水仙碱0. 00 012 g/kg组、血府逐瘀汤8. 0 g/kg组、茵栀黄9. 0 g/kg组、血府逐瘀汤联合茵栀黄组(4. 0 g/kg+4. 5 g/kg),每组10只,模型对照组18只。除正常对照组外,其余各组大鼠均按1. 5 ml/kg腹腔注射0. 5%DMN溶液,隔天1次;造模同时用药组按5 ml/kg灌胃相应体积药物进行干预,共60 d。末次给药后,禁食不禁水16 h,摘除眼球取血,采集肝组织。采用生化法检测血清中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)以及肝组织中丙二醛(MDA)、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GSH-Px)的活性或水平;ELISA法检测血清中层粘连蛋白(LN)、透明质酸(HA)、Ⅲ型前胶原(PCⅢ)以及肝组织中肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)和白介素-1β(IL-1β)的含量;Western blot检测肝组织中转化生长因子-β1(TGF-β1)、α-平滑肌肌动蛋白(α-SMA)的表达水平;以Masson染色法验证肝组织损伤程度。结果:与正常对照组相比,模型对照组肝组织SOD、GSH-Px活力显著降低,其余检测指标均显著上升(P <0. 01)。与模型对照组相比,除血府逐瘀汤组对血清HA、肝组织MDA、SOD、GSH-Px的含量或活性变化无统计学意义外,其余各给药组,特别是血府逐瘀汤4 g/kg联合茵栀黄4. 5 g/kg组对大鼠肝组织SOD、GSH-Px活性明显增强和其余检测指标的水平降低程度尤为明显(P <0. 05或P <0. 01)。结论:血府逐瘀汤和茵栀黄均对DMN致肝纤维化大鼠具有保护作用,且在剂量减半情况下,联合应用二者的综合疗效优于或与单独应用相当。

Objective: To study the protective effect of Xuefuzhuyu Decoction combined with Yinzhihuang on hepatic fibrosis induced by DMN in rats and its mechanism. Methods: Sixty-eight male SD rats were randomly divided into 6 groups according to their body weight: the control group,Colchicine group( 0. 00012 g/kg),Xuefuzhuyu Decoction group( 8. 0 g/kg),Yinzhihuang group( 9. 0 mg/kg),Xuefuzhuyu Decoc-tion combined with Yinzhihuang group( 4. 0 g/kg + 4. 5 mg/kg),10 rats in each group,and 18 rats in the model group. Except for the control group,rats in other groups were intraperitoneally injected with 1. 5 ml/kg 0. 5 % DMN every other day. While the models were being established,rats were treated with 5 ml/kg of the corresponding drugs for 60 days. After the last administration,fasting for 16 hours,the eyeballs were removed,and the liver tissues were collected. The levels or activities of ALT,AST in the serum and MDA,SOD and GSH-Px in liver tissues were detected by biochemical method. The contents of LN,HA,PCIII in serum and TNF-α,IL-6,IL-1β in liver tissues were detected by ELISA. The expression levels of α-SMA and TGF-β1 in liver tissues were detected by Western blot. Masson staining was used to verify the extent of liver tissue damage. Results: Compared with the control group,the detection indexes SOD and GSH-Px in the model group were significantly decreased,while the other indexes were significantly increased. Compared with the model group,no significant change was observed in the contents or activities of HA,MDA,SOD and GSH-Px in Xuefuzhuyu decoction group. The activities of SOD and GSH-Px in Yinzhihuang group and combined group were increased,while the levels of other detection indexes were decreased in Yinzhihuang group and combined group,in different degrees. And the effect in combined group showed significant difference. Conclusions: Xuefuzhuyu Decoction and Yinzhihuang has the protective effect on hepatic fibrosis induced by DMN in rats. At the half dose,the effect of combined use is better or equivalent to single use.