慢性阻塞性肺疾病早期远端肺小动脉重塑及相关NOX4表达水平研究  

Distal Pulmonary Arteriole Remodeling and Related NOX4 Expression in the Early Stage of Chronic Obstructive Pulmonary Disease

作　　者：高超[1,2] 郭晓桐 郝斌威[4] 曹霞 范玉春 陈娟[3] GAO Chao;GUO Xiaotong;HAO Binwei;CAO Xia;FAN Yuchun;CHEN Juan(Ningxia Medical University,Yinchuan 750004,China;Yinchuan Prison Hospital,Ningxia Hui Autonomous Region,Yinchuan 750004,China;Department of Respiratory and Critical Care Medicine,the General Hospital of Ningxia Medical University,Yinchuan 750004,China;Department of Respiratory and Critical Care Medicine,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences,Taiyuan 030000,China)

机构地区：[1]宁夏医科大学,银川750004 [2]宁夏回族自治区银川监狱医院,银川750004 [3]宁夏医科大学总医院呼吸与危重症医学科,银川750004 [4]山西白求恩医院山西医学科学院呼吸与危重症医学科,太原030000

出　　处：《宁夏医科大学学报》2022年第2期133-140,共8页Journal of Ningxia Medical University

基　　金：国家自然科学基金项目(81760004);呼吸疾病国家重点实验室2019年度开放课题(SKLRD-OP-201903);宁夏自然科学基金重点项目(2020AAC02001);第四批宁夏青年科技人才托举工程(TJGC2019101);宁夏医科大学校级科研项目(XM2020149)。

摘　　要：目的研究慢性阻塞性肺疾病(COPD)早期远端肺小动脉重塑情况及还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶4(NOX4)在肺小动脉的表达水平。方法收集2018年9月至2020年3月宁夏医科大学总医院收治的需行手术治疗的肺部肿瘤患者,根据术前肺功能及入组标准将受试者分为COPD组(24例)及对照组(20例),所有受试者术前均行心脏彩超检查以明确有无肺动脉高压。(1)术中取病变周围正常的肺组织,使用а-平滑肌肌动蛋白抗体(а-SMA)标记肺动脉平滑肌细胞。采用Image-Pro Plus software 6.0(IPP)图像分析软件测量远端肺小动脉(血管横断面外径为100~500μm)血管平滑肌层厚度(WT)、血管外径、血管平滑肌层面积(WA)、血管总面积;采用免疫组织化学法、蛋白免疫印迹法(Western blot)检测目的蛋白表达,应用IPP软件半定量计算目的蛋白平均光密度(AOD)值。(2)培养人原代肺动脉平滑肌细胞(HPASMC),使用重组人转化生长因子-β1(TGF-β1,2 ng·mL^(-1))蛋白作用于HPASMC,以检测目的蛋白表达。结果COPD组患者肺小动脉WT、平滑肌层厚度占血管外径的百分比(WT%)、平滑肌层面积占血管总面积百分比(WA%)均较对照组增高(P均<0.05)。COPD组肺小动脉平滑肌Ⅰ型胶原蛋白(Collagen I)、Ⅳ型胶原蛋白(CollagenⅣ)、层粘连蛋白(LN)、纤维连接蛋白(FN)、а-SMA及NOX4 AOD值均较对照组增强(P均<0.05),COPD组患者肺组织а-SMA、NOX4蛋白表达较对照组增强(P均<0.05)。肺动脉重塑指标WA%、WT%与第一秒用力呼气容积/用力肺活量(FEV1/FVC)、第一秒用力呼气容积占预计值百分比(FEV1%pred)均呈负相关(P均<0.05)。肺小动脉平滑肌CollagenⅠ、CollagenⅣ、LN、FNAOD值与FEV1/FVC、FEV1%pred均呈负相关(P均<0.05)。免疫细胞化学法、免疫细胞荧光法检测TGF-β1(2 ng·mL^(-1))作用于HPASMC 24 h后,NOX4、а-SMA、CollagenⅠ蛋白AOD值较未作用前增高(P均<0.05)。结论COPD患者早期存在以平Objective To investigate the remodeling of distal pulmonary arterioles and the expression level of reduced nicotinamide adenine dinucleotide phosphate oxidase 4(NOX4)in pulmonary arterioles in the early stage of chronic obstructive pulmonary disease(COPD).Methods Patients with lung tumors requiring surgical treatment treated in General Hospital of Ningxia Medical University from September 2018 to March 2020 were collected.According to the preoperative lung function and inclusion criteria,the subjects were divided into COPD group(24 cases)and control group(20 cases).All participants underwent color Doppler ultrasound before operation to determine whether there was pulmonary hypertension.(1)normal lung tissue around the lesion was taken during operation and usedа-smooth muscle actin antibody(а-SMA)labeled pulmonary artery smooth muscle cells.Image-Pro Plus software 6.0(IPP)image analysis software was used to measure the distal pulmonary arterioles(with a cross-sectional outer diameter of 100-500μm)vascular smooth muscle layer thickness(WT),vascular outer diameter,vascular smooth muscle layer area(WA)and total vascular area;the expression of target protein was detected by immunohistochemistry and Western blot,and the average optical density(AOD)of target protein was semi quantitatively calculated by IPP software.(2)human primary pulmonary artery smooth muscle cells(HPASMC)were cultured,using recombinant human transforming growth factor-β1(TGF-β1,2 ng·mL^(-1))protein acted on HPASMC to detect the expression of target protein.Results The WT of pulmonary arteriole,the percentage of smooth muscle thickness in the total vascular diameter(WT%)and the percentage of smooth muscle thickness in the total vascular area(WA%)in the COPD group were higher than those in the Control group(P<0.05).The protein expressions of AOD value collagenⅠ,collagenⅣ,laminin,fibronectin,a-SMA and NOX4 of pulmonary arteriolar smooth muscle of COPD group were higher than those of the control group(P all<0.05).The results of Western Blot s

关 键 词：慢性阻塞性肺疾病 肺小动脉重塑 还原型烟酰胺腺嘌呤二核苷酸磷酸氧化酶4 肺动脉高压 细胞外基质 

分 类 号：R563[医药卫生—呼吸系统]