苦参碱对肺纤维化大鼠核因子E2相关因子2、血红素氧合酶-1的作用影响

肺纤维化（pulmonary fibrosis，PF）是特发性肺纤维化、结节病、尘肺、过敏性肺炎、药物和放射导致的纤维化以及胶原血管病致纤维化肺泡炎等慢性肺部疾病的终末病理过程。在肺纤维化的形成机制中，氧化应激的作用机制成为主流，指氧化与抗氧化的失平衡状态。氧化应激促进TNF-α、IL-1、IL-8等前炎性介质表达的同时，提高具有保护性的γ-GCS、MnSOD、GST等[1]抗氧化基因的表达。核因子E2相关因子2（nuclear factor-erythroid2-related factor2，Nrf2）属于Cap-n Collar家族，它在抗氧化剂应激元件（ARE）介导的抗氧化基因表达中起重要的作用。血红素氧合酶-1（heme oxygenase-1，HO-1），谷胱甘肽-S-转移酶族（glutathione S-transferases，GSTs）和NAD（P）H醌氧化还原酶1[NAD（P）H quinoneoxi-doreductase1，NQO1]是基Ⅱ相酶主要组成部分。HO-1是催化血红素生成CO、胆红素及铁离子的限速酶，其保护作用通过其催化血红素分解的三种产物进行。HO-1的保护作用还可能与降低Caspase-3活性以对抗细胞凋亡和抑制TNF-α水平有关[2]。前期研究已经表明，Nrf2在COPD和哮喘肺中抗氧化应激中处于核心地位[2-3]。但是，Nrf2及其下游调控的HO-1在大鼠肺纤维化表达还有待于进一步研究。

苦参碱对肺纤维化大鼠核因子E2相关因子2、血红素氧合酶-1的作用研究影响

罗庆凯1陈伟2何振华2▲张秀峰2李炽观1郭春芳3

1.湖南省郴州市第一人民医院呼吸内科，湖南郴州423000；

2.南华大学附属第二医院呼吸内科，湖南衡阳421001；

3.中南大学卫生部肝胆肠外科研究中心，湖南长沙410008

[摘要]目的观察博来霉素诱导的大鼠肺纤维化中核因子E2相关因子2（Nrf2）、血红素氧合酶-1（HO-1）的表达，探讨苦参碱对肺纤维化的作用及Nrf2、HO-1表达的影响。方法SD大鼠120只，随机分为五组：正常对照组（C）、模型组（M）、泼尼松处理组（P）、苦参碱50处理组（M50）、苦参碱100处理组（M100）。计算肺系数，病理学检查肺组织，免疫组化法（SP）分析每组肺组织中的Nrf2、HO-1蛋白水平，RT-PCR检测大鼠肺组织中Nrf2、HO-1mRNA表达。

结果造模后M、P、M50、M100组的Nrf2、HO-1表达高于C组（P＜0.05）。第7、14天时M组Nrf2、HO-1mRNA表达比P、M50、M100组低（P＜0.05）；第28天M、P、M50、M100组Nrf2、HO-1mRNA表达比第7、14天的表达偏低，第7天表达最高（P＜0.05）。直线相关分析显示Nrf2mRNA表达与HO-1mRNA呈正相关（P＜0.05）。结论苦参碱有抑制博来霉素诱导的肺纤维化的作用，其机制可能与早期上调Nrf2、HO-1表达、抑制氧化应激反应有关。

[关键词]肺纤维化；苦参碱；核因子E2相关因子2；血红素氧合酶-1；博来霉素

[中图分类号]R284[文献标识码]A[文章编号]1673-7210（2012）07（c）-0018-04

Study on the role and mechanism of the matrine effected with Nrf2,HO-1 in rats with pulmonary fibrosis

LUO Qingkai1CHEN Wei2HE Zhenhua2▲ZHANG Xiufeng2LI Zhiguan1GUO Chunfang3

1.Department of Respiratory Medicine,Chenzhou First People's Hospital of Hu'nan Province,Chenzhou423000,China;

2.Department of Respiratory Medicine,the Second Affiliated Hospital of Nanhua University,Hu'nan Province,Hengyang

421001,China;3.Hepatobiliary and Enteric Surgery Research Center of Ministry of Health of Central South University, Hu'nan Province,Changsha410008,China

[Abstract]Objective To observe the expression of Nrf2,HO-1in rats which had pulmonary fibrosis induced by bleomycin.Study the role of matrine and the expression of the Nrf2,HO-1which intervened by the matrine.Methods120 SD rats were randomly divided into5groups:normal control group(C),model group(M),prednisone treatment group(P), matrine50treatment group(M50)and the matrine100treatment group(M100).The coefficients of the lung were calculat-ed and the lung tissue was checked with pathological examination.Protein level of Nrf2,HO-1at different time in each group were analyzed by immunohistochemistry(SP),while the mRNA level of Nrf2,HO-1were deceted by RT-PCR.Re-sults Quantity of Nrf2,HO-1expressed in group M,group P,group M50and group M100were higher than those in group

C.The expression level of Nrf2,HO-1mRNA in group M was lower than those in group P,M50,M100(P＜0.05)at the

7th and14th day.And the expression level of Nrf2,HO-1mRNA in group M,P,M50,M100at the28th day was lower than the7th and14th day respectively,while the expression at the7th day is the highest(P＜0.05).The linear correlation analysis showed that the quantity of Nrf2mRNA was positively correlated with HO-1mRNA(P＜0.05).Conclusion The pulmonary fibrosis induced by bleomycin in rats can be controled by matrine at some degree,and the mechanism may be related with the upregulating of the Nrf2,HO-1expression at the early times and controling the oxidative stress reaction.

[Key words]Pulmonary fibrosis;Matrine;Nuclear factor-erythroid2-related factor2;Heme oxygenase-1;Bleomycin

▲通讯作者

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中国医药导报CHINA MEDICAL HERALD