川芎嗪对土三七诱导的小鼠肝小静脉闭塞病治疗机制的研究

摘要

Background:Hepatic veno-occlusive disease( HVOD) is a disease characterized by hepatomegaly,jaundice, ascites,weight gain and lack of effective treatment currently. Our prophase research showed that ligustrazine had therapeutic effect on Sedum aizoon induced HVOD in mice. Aims:To investigate the mechanism of therapeutic effect of ligustrazine on Sedum aizoon induced HVOD in mice. Methods:A total of 115 mice were randomly divided into 4 groups:mice in group A were intragastrically administrated with 30 mg·kg-1 ·d-1 Sedum aizoon to induce HVOD and served as model group;mice in group B were given 30 mg·kg-1 ·d-1 Sedum aizoon + 100 mg·kg-1 ·d-1 ligustrazine and served as low dose ligustrazine intervention group;mice in group C were given 30 mg·kg-1 ·d-1 Sedum aizoon + 200 mg·kg-1 ·d-1 ligustrazine and served as high dose ligustrazine intervention group;mice in group D were given 30 mg·kg-1 ·d-1 PBS and served as normal control group. After 30 days,all the mice were sacrificed. HE staining and Masson staining were performed for histological examination. The mRNA and protein expressions of tissue factor(TF),nuclear factor(NF)-κBp65 and early growth response factor( Egr)-1 in liver tissue were determined by RT-PCR and Western blotting, respectively. Results:HE staining and Masson staining histological examination showed that ligustrazine could obviously ameliorate the pathological injury of liver tissue in HVOD mice. Compared with group D,the mRNA and protein expressions of TF,NF-κBp65,Egr-1 were significantly increased in group A( P < 0. 05). After intervention with ligustrazine,the mRNA and protein expressions of TF,NF-κBp65,Egr-1 were significantly decreased( P < 0. 05), especially in group C,and no significant differences were found between group C and group D(P > 0. 05). Conclusions:Ligustrazine has therapeutic effect on HVOD,the possible mechanism is that ligustrazine could interrupt the activation of coagulation system by reducing the expression of TF via down regulating the expressions of NF-κBp65 and Egr-1,especially in high dose ligustrazine group.%背景：肝小静脉闭塞病（HVOD）临床上以肝肿大、黄疸、腹水和体重增加为特征，目前尚缺乏有效的治疗手段。本课题前期研究发现川芎嗪对土三七诱导的小鼠 HVOD 具有治疗作用。目的：探讨川芎嗪对土三七诱导的小鼠HVOD 的治疗机制。方法：将115只小鼠随机分为土三七组（土三七浓缩煎液30 g·kg-1·d-1灌胃）、低剂量川芎嗪干预组（土三七浓缩煎液30 g·kg-1·d-1＋川芎嗪100 mg·kg-1·d-1灌胃）、高剂量川芎嗪干预组（土三七浓缩煎液30 g·kg-1·d-1＋川芎嗪200 mg·kg-1·d-1灌胃）和正常对照组（PBS 30 g·kg-1·d-1灌胃），30 d 后处死所有小鼠。行HE 染色和 Masson 染色，以 RT-PCR 法和蛋白质印迹法分别检测肝组织中组织因子（TF）、核因子（NF）-κBp65、早期生长反应因子-1（Egr-1）mRNA 和蛋白表达。结果：HE 染色和 Masson 染色结果显示川芎嗪可明显改善 HVOD 小鼠肝组织病理损伤。土三七组 TF、NF-κBp65和 Egr-1 mRNA 和蛋白表达显著高于正常对照组，组间差异有统计学意义（P ＜0.05）；川芎嗪干预后，TF、NF-κBp65和 Egr-1 mRNA 和蛋白表达均不同程度下降（P ＜0.05），以高剂量组下降更为明显，而高剂量川芎嗪干预组与正常对照组相比差异无统计学意义（P ＞0.05）。结论：川芎嗪可能通过下调 NF-κBp65和 Egr-1表达降低 TF 水平，阻止凝血系统活化，从而有效治疗 HVOD，且高剂量川芎嗪的疗效更为确切。