血红素加氧酶1调节γ谷氨酰半胱氨酸合成酶的表达在慢性阻塞性疾病大鼠肺组织中的作用  

Effect of Heme Oxygenase-1 on Expression of γ-Glutamyl-cysteine Synthetase in Chronic Obstructive Pulmonary Disease Rats

作　　者：王梅芳[1,2] 戴爱国 刘玉全[2] 

机构地区：[1]湖南省老年医院.湖南省老年医学研究所呼吸疾病研究室,长沙410001 [2]湖北郧阳医学院附属太和医院呼吸内科,湖北十堰442000

出　　处：《中国生物化学与分子生物学报》2008年第12期1158-1164,共7页Chinese Journal of Biochemistry and Molecular Biology

摘　　要：血红素加氧酶1(heme oxygenase-1,HO-1)是催化血红素降解酶类中最易被诱导的一种,增加HO-1表达有抗氧化的细胞保护作用;而γ-谷氨酰半胱氨酸合成酶(gamma-glutamyl cysteine synthetase,γ-GCS)是肺组织中最重要的抗氧化酶之一,通过免疫组化、免疫印迹、逆转录-聚合酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)技术研究慢性阻塞性肺疾病(chronic obstructive pulmonary disease,COPD)大鼠肺内HO-1、γ-GCS mRNA及其蛋白质表达,并检测HO-1活性,同时利用HO-1诱导剂氯高铁血红素(hemin)和其活性抑制剂锡原卟啉(SnPP)对其进行干预,在整体水平探讨HO-1在COPD中对肺功能及γ-GCS表达的影响.结果显示,氯高铁血红素组肺功能较COPD组及SnPP组显著改善;氯高铁血红素组及SnPP组Ho-1 mRNA及其蛋白质的表达较COPD组升高;氯高铁血红素组HO-1的活性较COPD升高而Snpp组HO-1的活性较COPD组降低;氯高铁血红素组γ-GCS mRNA及其蛋白质表达水平高于SnPP及COPD组;线性回归分析显示,HO-1蛋白质表达水平和活性影响γ-GCS表达.结果表明,HO-1可通过诱导γ-GCS的表达在COPD中起保护作用;而HO-1可能通过酶活性依赖和非酶活性依赖两种途径上调γ-GCS基因表达.Heine oxygenase-1 （HO-1） is an inducible enzyme that catalyzes hemin to generate bilirubin, carbon monoxide and ferri. Because enhanced expression of HO-1 provides a anti-oxidative effect and confers eytoprotection, we examined whether HO-1 overexpression induced by hemin in the lung would prevent rat pulmonary emphysema induced by cigaret smoking （CS） combined with lipopolysaceharide（LPS） and explored its possible molecular mechanisms. SnPP was used to depress HO-1 activity. Reverse transcription -polymerase chain reaction （RT-PCR）,immunohistochemistry and Western blot techniques were used. The treatment with hemin, which up-regulated production of HO-1 and activity of HO-1 in chronic obstructive pulmonary disease （COPD） rat lung, attenuated the CS combined with LPS induced enlargement of alveoli and pulmonary functions aggravation. It also up-regulated the expression of γ-GCS, one of the most important anti-oxidative gene in lung tissue. The treatment with SnPP, which up-regulated production of HO-1 but degradation activity of HO-1 in COPD rat lung, had no effect on pulmonary function aggravation and the expression of γ-GCS.Theliner regression showed that both HO-1 protein level and its activity have effects on the expression of γ-GCS. The results imply that hemin-induced HO-1 overexpression suppressed emphysema caused by CS combined with LPS through up-regulating the expression of γ-GCS in enzymatic activities though dependent and independent pathways.

关 键 词：阻塞性肺疾病 血红素加氧酶1 Γ-谷氨酰半胱氨酸合成酶 氧化应激 

分 类 号：R563.3[医药卫生—呼吸系统]