复方肝毒清对二甲基亚硝胺诱导肝纤维化大鼠肝组织细胞因子表达的影响

Effects of compound Ganduqing decoction （复方肝毒清） on mRNA expression of cytokine in rats of N-Nitrosodimethylamine induced liver fibrosis

目的探讨复方肝毒清对二甲基亚硝胺（DMN）诱导肝纤维化大鼠肝组织转化生长因子-β1（TGF-β1）、诱导型一氧化氮合酶（iNOS）、肿瘤坏死因子a（TNF—a）的mRNA表达的影响。方法采用每周连续3d腹腔注射DMN10／ul／kg共4周制备大鼠肝纤维化模型（70只），并设对照组（10只）。第2周开始，将70只模型大鼠按随机数字表法分为模型组、秋水仙碱组、氯化钆（GdCl3）组以及复方肝毒清高、低剂量组，后4组分别灌胃秋水仙碱0．01mg／kg、腹腔注射GdCl3 40mg／kg及灌胃中药复方肝毒清20g／kg、5g／kg；对照组及模型组均灌胃生理盐水10ml／kg；均每日1次，连用21d。采用定量聚合酶链反应（PCR）检测肝组织TGF—β1、iNOS、TNF-a的FiRRNA表达。结果中药复方肝毒清不仅能减轻肝组织铁的沉积，抑制胶原纤维的增生，减少假小叶的形成；而且能不同程度下调TGF—β1、iNOS、TNF—a的mRNA表达，以高剂量组较显著，与模型组比较差异有统计学意义（TGF—β1mRNA：6．32±1．19比16．15±2．39，iNOSmRNA：18．73±2．13比43．15±6．39，TNF-amRNA：3．69±1．22比8．68±2．13，均P〈0．01）。结论铁沉积于库普弗细胞在肝纤维化发生发展过程中发挥重要作用，复方肝毒清的抗肝纤维化机制可能与其调节体内铁的代谢有关。

Objective To investigate the effects of compound Ganduqing decoction （复方肝毒清） on mRNA expressions of transforming growth factor-β1 （TGF-β1）, inducible nitric oxide synthase （iNOS）, tumor necrosis factor-a （TNF-a） in rats of N-Nitrosodimethylamine （DMN） induced liver fibrosis. Methods Rat liver fibrosis models were established by DMN intraperitoneal injection at a dose of 10 ul/kg daily, for 3 consecutive days each week for 4 weeks, and blank controlgroup （10 rats） was made. Fromthe second week, 70 rats for liver fibrosis models were randomly divided into model group, colchicine group, GdCl3 group, compound Ganduqing high-dose and low-dose groups. Rats of the last four groups were treated by gastric infusion at the dosage of colchicine 0.01 mg/kg, intraperitoneal injection of GdCl3 40 mg/kg, gastric perfusion of compound Ganduqing decoction at the dosage of 20 g/kg and 5 g/kg, respectively. The model group and blank control group were both infused gastrically with normal saline at 10 ml/kg. All groups were treated once daily for 21 days. Quantitative polymerase chain reaction （PCR） was applied to detect expressions of TGF-β1, iNOS and TNF-a mRNA. Results Compound Ganduqing not only could reduce iron deposition, prevent the proliferation of collagen fibers and decrease pseudolobule formation in liver, but also could down-regulate TGF-β1, iNOS and TNF-a mRNA expressions of liver tissue, which were more considerable in the Ganduqing high-dose group. Compared with the model group, there were statistical significant differences （TGF-β1 mRNA： 6.32±1. 19 vs. 16.15±2.39, iNOS mRNA： 18.73±2.13 vs. 43.15±6.39, TNF-a mRNA： 3.69± 1.22 vs. 8.68±2.13, all P〈0.01）. Conclusion Iron deposition in Kupffer cells may play an important role in the genesis and development of liver fibrosis. Compound Ganduqing can ameliorate liver fibrosis, whose anti-fibrosis mechanism might be related to Ganduqing regulatory action on iron metabolism of liver tissues.