摘要
目的:探讨过氧化物酶体增生物激活受体γ(PPARγ)对慢性阻塞性肺疾病(COPD)大鼠肺γ-谷氨酰半胱氨酸合成酶(γ-GCS)活性及表达的影响,及在COPD发病中的作用。方法:用每天熏香烟和2次气管内滴入脂多糖(LPS)法制作大鼠COPD模型,同时利用PPARγ激活剂罗格列酮(RGZ)对其进行干预,测定3组大鼠肺功能和病理变化结果;并检测3组大鼠肺内ROS含量和γ-GCS活性;应用免疫组化、免疫印迹(Western blot-ting)、原位杂交和逆转录-聚合酶链反应(RT-PCR)方法检测PPARγ、γ-GCS mRNA及蛋白在3组大鼠肺组织中的表达情况。结果:RGZ干预组大鼠肺功能指标(FEV0.3、FEV0.3/FVC%与PEF)均较COPD组明显好转;光镜下COPD组肺组织病理变化符合COPD的特征性改变,RGZ干预组肺组织病理变化较COPD组显著减轻;RGZ干预组ROS含量较COPD组显著减少,而γ-GCS活性较COPD组升高;PPARγ、γ-GCSmRNA及其蛋白质表达在COPD组均较对照组明显增高(均P<0.01),而在RGZ干预组均较COPD组增高(均P<0.05);直线相关分析显示PPARγ蛋白与γ-GCS活性呈正相关(r=0.634,P<0.01),与ROS含量无明显相关性(r=0.214,P>0.05);PPARγ蛋白与γ-GCS蛋白及mRNA表达呈正相关(r=0.553、r=0.442,均P<0.01)。结论:RGZ活化PPARγ可减轻COPD氧化/抗氧化失衡程度,对COPD的防治起重要作用;另外,PPARγ可能通过减少肺内ROS的产生,增强γ-GCS活性及其基因表达而在COPD中起重要的抗氧化保护作用。
AIM： To explore the effects of peroxisome proliferator- activated receptor γ （PPARγ） on the activity and expression of γ - glutamylcysteine synthetase （ γ - GCS）, and its role in rats with chronic obstructive pulmonary disease. METHODS： COPD model was established by the method of combining fumigation and lipopolysaccharide （LPS） intra - tracheal dripping. Meanwhile, some of the COPD rats were administered with rosiglitazone （ RGZ）, a PPARγ activator. The pulmonary function and the pathological changes were determined. ROS content and γ - GCS activity in lung tissues were detected. The levels of PPARγ, γ - GCS mRNA and protein expression in lung tissues were measured by immunohistochemistry, Western blotting, in situ hybridization （ISH） and reverse transcription - polymerase chain reaction （ RT -PCR）. RESULTS： The pulmonary function （FEV0.3 , FEV0.3/FVC% , PEF） were significantly improved in RGZ group compared to COPD group. Under light microscope, lung pathological changes in COPD group eonfoimed to pathological features of COPD. The pathological changes of lung tissue were obviously reduced in RGZ group compared to COPD group. In RGZ group, ROS content was obviously reduced and γ- GCS activity significantly increased compared to COPD group. Protein and mRNA expressions of PPARγ and γ- GCS in COPD group significantly higher than those in control group （ all P 〈 0. O1 ） , and those in RGZ group was markedly increased compared to COPD group （ all P 〈 0. 05 ）. Linear correlation analysis showed that PPARγ protein was positively correlated with γ - GCS activity （ r = 0. 634, P 〈 0. 01 ）, and was no significantly correlated with ROS content （ r = 0. 214, P 〉 0. 05 ）. PPARγ protein was positively correlated with γ-GCS protein and mRNA （r = 0. 553, r = 0. 442, all P 〈 0. 01 ）. CONCLUSION： PPARγ activation by RGZ reduces the extent of COPD oxidant/antioxidant imbalance, which plays an important role in the prevention and treatment of COPD. In addition, PPARγ/may play an important antioxidant protection role by reducing ROS production, and increasing activity and gene expression of γ- GCS in the lung.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2010年第2期302-308,共7页
Chinese Journal of Pathophysiology
基金
湖南省科技计划资助项目(No.05JT1023
No.2007SK3080)
湖南省科技计划重点资助项目(No.2007FJ3016
No.2008FJ4206)
湖南省医药卫生科研课题资助项目(No.B2005125
No.B2007166)
湖南省自然科学基金资助项目(No.09JJ5021)
