RhoA/Rho激酶1在慢性阻塞性肺疾病患者肺小静脉、肺小动脉及小气道平滑肌的表达研究  

ROCK1 expression of the smooth muscles of the pulmonary venules,pulmonary arterioles and small airways in patients with chronic obstructive pulmonary disease

作　　者：郭晓桐[1] 高超 崔节达 曹霞 陈娟[1] Guo Xiaotong;Gao Chao;Cui Jieda;Cao Xia;Chen Juan(Department of Respiratory and Critical Care Medicine,Ningxia Medical University General Hospital,Yinchuan 750004,China;Ningxia Xingqing Prison(Internal Medicine Ward,Central Hospital of Ningxia Prison Administration),Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院呼吸与危重症医学科,银川750004 [2]宁夏兴庆监狱(宁夏监狱管理局中心医院内科病区),银川750004

出　　处：《国际呼吸杂志》2022年第12期901-909,共9页International Journal of Respiration

基　　金：国家自然科学基金(81760004);宁夏自然科学基金重点项目(2020AAC02001);宁夏医科大学校级科研项目(XM2020149);第四批宁夏青年科技人才托举工程(TJGC2019101)。

摘　　要：目的探讨RhoA/Rho激酶1(ROCK1)参与细胞外基质(ECM)沉积对慢性阻塞性肺疾病(COPD)患者肺小静脉、肺小动脉及小气道平滑肌重塑的病理生理意义。方法本研究为病例对照研究。采用非随机抽样法,分析2019年10月至2020年12月宁夏医科大学总医院收治的因原发性肺肿瘤而接受肺切除术患者的肺组织,结合患者肺功能及临床病史分为COPD组(n=13),对照组(n=12)。使用α-平滑肌肌动蛋白(α-SMA)抗体鉴定平滑肌细胞,选取高倍视野下结构完整的肺小静脉及肺小动脉(血管横断面外径<500μm)、小气道(气道横断面外径<2000μm),使用Image-Pro Plus测量肺小静脉、肺小动脉血管外径,小气道基底膜周长,计算肺小静脉、肺小动脉及小气道平滑肌层面积占管腔横断面面积百分比(WA%)作为评估平滑肌重塑的指标。采用免疫组织化学及蛋白免疫印迹法检测目的蛋白的表达。分析形态学和临床资料之间的相关性。结果COPD组肺小静脉、肺小动脉、小气道WA%均较对照组增高(P值均<0.05)。免疫组织化学检测显示COPD组肺小静脉、肺小动脉、小气道平滑肌细胞α-SMA、纤维连接蛋白(FN)、ROCK1表达中位平均光密度(AOD)值均较对照组增高(P值均<0.05)。蛋白免疫印迹法显示,COPD组肺组织α-SMA、FN、ROCK1蛋白表达灰度值分别为[(1.31±1.07)、(0.55±0.23)、(0.64±0.20)],均较对照组增高,分别为[(0.59±0.37)、(0.26±0.16)、(0.33±0.18),t值分别为2.30、3.86、4.06,P值均<0.05]。相关性分析显示,第1秒用力呼气容积/用力肺活量、第1秒用力呼气容积占预计之百分比与肺小静脉、肺小动脉、小气道WA%均呈负相关,与肺小静脉、肺小动脉、小气道平滑肌细胞ROCK1表达AOD值均呈负相关(P值均<0.05)。COPD组患者外周血中性粒细胞绝对值及超敏C反应蛋白较对照组升高(P值均<0.05)。结论轻中度稳定期COPD患者的肺小静脉、肺小动脉、小气道存在以平滑肌�Objective To investigate the pathophysiological significance of RhoA/Rho kinase 1(ROCK1)involvement in extracellular matrix(ECM)deposition in the remodeling of pulmonary venules,pulmonary arterioles and small airways smooth muscle in patients with chronic obstructive pulmonary disease(COPD).Methods This was a case-control study.The non-random sampling method was used in this study.Lung tissues of patients who underwent pneumonectomy for primary lung tumor admitted to Ningxia Medical University General Hospital from October 2019 to December 2020 were analyzed.Combined with lung function and clinical history,the patients were divided into COPD group(n=13)and control group(n=12).The smooth muscle cells were identified usingα-smooth muscle actin(α-SMA)antibody.Distal pulmonary venules and pulmonary arterioles(vessel cross section diameter<500μm)and small airways(airway cross section diameter<2000μm)with complete structure were selected from high magnification field of view.The external diameter(ED)of the pulmonary venules,pulmonary arterioles and the perimeter of the basement membrane(Pbm)of the small airway were measured using Image-Pro Plus and the percentage of smooth muscle layer area to cross-sectional area(WA%)of pulmonary venules,pulmonary arterioles and small airways were calculated as the evaluation indexes of smooth muscle remodeling.Immunohistochemical and Western blot were used to detect the difference of protein expression between the two groups.The correlation between morphology and clinical data was analyzed.Results The WA%of pulmonary venules,pulmonary arterioles and small airways in COPD group were higher than those in control group(all P<0.05).Immunohistochemical analysis showed that the Average Optical Density(AOD)ofα-SMA,fibronectin(FN)and ROCK1 expression in pulmonary venules,pulmonary arterioles and small airways smooth muscle in COPD group were were higher than those in control group(all P<0.05).Western blotting showed that the gray levels ofα-SMA,FN,and ROCK1 protein expression in COPD g

关 键 词：肺疾病 慢性阻塞性 细胞外基质 RHOA/RHO激酶 

分 类 号：R563.9[医药卫生—呼吸系统]