CAJ | 학술논문

目的探讨内源性气体信号分子硫化氢在慢性阻塞性肺疾病(COPD)患者肺组织中的表达变化.方法收集24例肺癌手术正常肺组织标本,包括非吸烟者11例,肺功能正常的吸烟者7例,COPD患者6例.组织切片HE染色后光镜下观察肺组织病理形态特征,检测肺组织中还原型谷胱甘肽(GSH)和超氧化物歧化酶(SOD)水平以反映氧化应激状态.采用Western blotting、Real-time PCR和免疫组织化学的方法检测内源性硫化氢合成酶蛋白水平和mRNA水平的表达变化.采用敏感硫电极测定肺组织匀浆中硫化氢水平.结果三组患者在年龄、身高、体质量方面差异无统计学意义,COPD组患者FEV1/FVC和FEV1占预计值百分比显著低于吸烟组和非吸烟组.COPD组和吸烟组肺组织中GSH和SOD均降低.与非吸烟组相比,吸烟组和COPD组肺组织中硫化氢合成酶-胱硫醚-γ-裂解酶(CSE)的蛋白表达分别减少35％和39％；COPD组CSE mRNA水平比非吸烟组增加了7.6倍；三组患者肺组织匀浆中硫化氢含量差异无统计学意义(P=0.27).免疫组织化学结果表明,CSE主要表达于气道平滑肌细胞、血管平滑肌细胞以及肺泡上皮细胞等细胞的细胞浆中,CSE在非吸烟组表达最强,呈棕褐色颗粒,在吸烟组和COPD组为棕黄色颗粒,且比非吸烟组表达明显减少.结论内源性硫化氢在COPD患者肺组织中表达受损.
목적 탐토내원성기체신호분자류화경재만성조새성폐질병(COPD)환자폐조직중적표체변화.방법 수집24례폐암수술정상폐조직표본,포괄비흡연자11례,폐공능정상적흡연자7례,COPD환자6례.조직절편HE염색후광경하관찰폐조직병리형태특정,검측폐조직중환원형곡광감태(GSH)화초양화물기화매(SOD)수평이반영양화응격상태.채용Western blotting、Real-time PCR화면역조직화학적방법검측내원성류화경합성매단백수평화mRNA수평적표체변화.채용민감류전겁측정폐조직균장중류화경수평.결과 삼조환자재년령、신고、체질량방면차이무통계학의의,COPD조환자FEV1/FVC화FEV1점예계치백분비현저저우흡연조화비흡연조.COPD조화흡연조폐조직중GSH화SOD균강저.여비흡연조상비,흡연조화COPD조폐조직중류화경합성매-광류미-γ-렬해매(CSE)적단백표체분별감소35％화39％；COPD조CSE mRNA수평비비흡연조증가료7.6배；삼조환자폐조직균장중류화경함량차이무통계학의의(P=0.27).면역조직화학결과표명,CSE주요표체우기도평활기세포、혈관평활기세포이급폐포상피세포등세포적세포장중,CSE재비흡연조표체최강,정종갈색과립,재흡연조화COPD조위종황색과립,차비비흡연조표체명현감소.결론 내원성류화경재COPD환자폐조직중표체수손.
Objective To investigate the expression of endogenous H2S in lung tissue from chronic obstructive pulmonary disease (COPD) patients.Methods Peripheral lung tissue samples were collected from 24 patients-11 nonsmokers,7 smokers who had normal lung function and 6 COPD patients.Histological changes of lungs were observed under light microscope.The content of glutathione (GSH)and the activity of superoxide dismutase (SOD) were detected to reflect oxidative stress.The protein and mRNA levels of endogenous H2 S synthetase were analyzed.The level of H2 S in lung tissue was analyzed by the sensitive sulphur electrode assay.Results FEV1/FVC and proportion of predicted FEV1 were significantly lower in patients with COPD than in smokers and nonsmokers.GSH and SOD were decreased in lung tissue of COPD and smokers.When compared with nonsmokers,the protein level of cystathionine-γ-lyase (CSE,the major enzyme producing H2S in the cardiovascular and respiratory system) was decreased in lung tissue from smokers and COPD patients by 35％ and 39％ respectively.However CSE mRNA was increased in COPD patients.There was no significant difference in H2S levels in lung tissue in each group.Immunohistochemistry showed that CSE was mainly expressed in bronchial and alveolar epithelial cells in nonsmokers' lung tissue.In smokers and COPD patients,CGL expression was decreased.Conclusions Endogenous H2S is impaired in patients with COPD.