RELMβ在COPD相关肺动脉高压患者发病中作用及机制研究

发布时间：2018-06-19 03:50

本文选题：肺动脉高压 + COPD　；参考：《南华大学》2015年硕士论文

【摘要】：目的:研究COPD相关肺动脉高压(pulmonary hypertension,PH)患者肺血管重塑病理过程中,抵抗素样分子β(resistin-like moleculeβ,RELMβ)及其介导的信号通路PI3K/Akt/m TOR在肺小动脉血管壁的表达情况,明确上述蛋白在肺血管重塑过程中的作用。通过检测COPD相关PH患者血浆中RELMβ、HIF-1a蛋白表达水平探讨其与肺动脉压力的相关性,从而为研究COPD相关PH提供新的理论依据。方法:实验共分为两个部分。第一部分收集因肺鳞癌行肺叶切除术患者的肺组织30例,通过血气分析、肺功能、多普勒超声心动图将其分为非COPD非PH组(对照组)、COPD非PH组(COPD组)、COPD相关PH组(PH组)。观察各组肺血管重塑情况,免疫组织化学染色法(IHC)检测RELMβ蛋白在肺组织的表达部位及水平,免疫蛋白印迹(Western blot,WB)检测RELMβ、PI3K、Akt、m TOR蛋白在患者肺小动脉血管壁上的表达情况,并探讨上述蛋白在肺血管重塑过程中的作用。第二部分将因COPD急性加重而入院治疗的80例患者通过血气分析、肺功能、多普勒超声心动图将其分为COPD相关PH组(PH组)、COPD非PH组(COPD组),并在体检科随机抽取行上述检查的健康志愿者40例为对照组,采集各观察组患者静脉血5ml,通过离心机分离出血浆,用ELISA法检测各组患者血浆中Relmβ、HIF-1a蛋白表达水平并探讨其与肺动脉压力的相关性。结果:1、显微镜下观察各组肺血管重塑(1)对照组显微镜下可见肺泡结构清晰,肺泡间隔完整,肺小动脉管壁较薄,血管腔未见狭窄。(2)COPD组较对照组相比,肺泡壁断裂、融合,可见肺大泡形成,肺泡结构破坏明显,肺小动脉血管壁增厚,血管腔狭窄明显。COPD组肺小动脉管壁厚度占外径的百分比(WT%)、肺小动脉管壁面积与血管总面积的比值(WA%)均高于对照组,差异有统计学意义(P0.05)。而肺小动脉管腔面积与管总面积的比值(LA%)低于对照组,差异有统计学意义(P0.05)。(3)PH组显微镜下可见肺大泡形成,其肺小动脉血管中膜层显著增厚,血管腔明显狭窄,该组WT%、WA%、LA%较COPD组比较,差异均存在显著统计学意义(P0.05)。2、免疫组织化学检测各组肺小动脉血管壁RELMβ表达免疫组织化学染色法结果显示:COPD组与对照组相比,RELMβ表达明显增强,差异具有统计学意义(P0.05);PH组则呈强阳性表达,与COPD组比较差异有统计学意义(P0.05)。3、Western blot检测各组患者肺组织内RELMβ、PI3K/Akt/m TOR信号通路蛋白表达Western blot结果显示:Relmβ在COPD组表达量显著高于对照组,结果具有统计学意义(P0.05);PH组Relmβ蛋白表达量高于COPD组,结果有统计学意义(P0.05);p-PI3K在COPD组表达量显著高于对照组,结果具有统计学意义(P0.05);p-PI3K在PH组表达量显著高于COPD组,结果具有统计学意义(P0.05);p-Akt308在COPD组表达量明显高于对照组,结果具有统计学意义(P0.05);Akt308在PH组表达量明显高于COPD组,结果具有统计学意义(P0.05);p-Akt473在COPD组表达量明显高于对照组,结果具有统计学意义(P0.05);p-Akt473在PH组表达量明显高于COPD组,结果具有统计学意义(P0.05);p-m TOR在COPD组表达量明显高于对照组,结果具有统计学意义(P0.05);p-m TOR在PH组表达量明显高于COPD组,结果具有统计学意义(P0.05)。直线相关分析显示,患者肺组织内Relmβ蛋白与WT%、WA%、PASP呈正相关,与LA%、动脉血氧分压(Pa O2)呈负相关。4、ELISA检测各组患者血浆中Relmβ、HIF-1a表达COPD组患者血浆中Relmβ表达量与对照组相比,差异具有统计学意义(P0.05);PH组患者血浆中Relmβ表达量与COPD组相比,具有显著统计学差异(P0.05);COPD组患者血浆中HIF-1a表达量与对照组相比,差异具有统计学意义(P0.05);COPD组患者血浆中HIF-1a表达量与PH组相比,差异具有统计学意义(P0.05)。直线相关分析显示,患者血浆中RELMβ蛋白表达与PASP、HIF-1a呈正相关,与Pa O2呈负相关,而患者血浆中HIF-1a蛋白表达与PASP呈正相关,与Pa O2呈负相关。1、COPD相关PH患者肺小动脉血管壁中Relmβ表达显著增高,并可能通过激活PI3K/Akt/m TOR信号通路,参与肺血管重塑,从而导致PH的形成。2、COPD相关PH患者血浆中Relmβ、HIF-1a表达显著增高,并与肺动脉压力呈正相关。结论:
[Abstract]:Objective: To investigate the expression of resistin-like molecule beta (RELM beta) and its mediated signaling pathway PI3K/Akt/m TOR in the vascular wall of pulmonary artery in the pathological process of pulmonary vascular remodeling in patients with COPD associated pulmonary hypertension (PH), and to clarify the role of these proteins in the process of pulmonary vascular remodeling. The correlation between RELM beta and HIF-1a protein expression in the plasma of COPD related PH patients was examined to provide a new theoretical basis for the study of COPD related PH. Methods: the experiment was divided into two parts. The first part collected 30 cases of lung tissue from patients with lung squamous cell carcinoma with pulmonary lobectomy, through blood gas analysis, lung function, and multip It was divided into non COPD non PH group (control group), non PH group COPD (group COPD), COPD related PH group (PH group). The pulmonary vascular remodeling was observed in each group. Immunohistochemistry staining method (IHC) was used to detect the expression of RELM beta protein in the lung tissue. The expression of the expression on the vascular wall of the pulmonary artery and the role of the above protein in the process of pulmonary vascular remodeling. Second part of the 80 patients who were hospitalized for the acute exacerbation of COPD were divided into COPD related PH group (group PH), COPD non PH group (group COPD), and random extraction in the physical examination department. 40 healthy volunteers were taken as the control group. The venous blood 5ml was collected from the observation groups and the blood plasma was separated by centrifuge. The level of Relm beta, HIF-1a protein expression in the plasma of each group was detected by ELISA and the correlation between the pulmonary pressure and the pulmonary artery pressure was investigated. Results: 1, the pulmonary vascular remodeling (1) in each group was observed under microscope (1). Microscopically, the alveolar structure was clear, the alveolar septum was complete, the wall of the pulmonary arteriole was thinner and the vascular cavity was not narrow. (2) compared with the control group, the alveolar wall was broken, the pulmonary bullae formed, the alveolar structure was damaged obviously, the pulmonary artery wall was thickened, the stenosis of the vascular cavity was obvious in the.COPD group, and the thickness of the tube wall of the pulmonary artery accounted for 100 percent of the diameter in the COPD group. The ratio of pulmonary arteriole wall area to total vascular area (WA%) was higher than that of the control group (WT%), and the difference was statistically significant (P0.05). The ratio of the lumen area to the total area of the pulmonary artery was lower than that of the control group (LA%), the difference was statistically significant (P0.05). (3) the formation of the pulmonary bullae was observed under the microscope of the group PH, and the membrane of the pulmonary arteriole was significant. Thickening, the vascular cavity was obviously narrowed, the group WT%, WA%, LA% compared with the COPD group, the difference was statistically significant (P0.05).2. The immunohistochemical staining of the RELM beta expression of the vascular wall of the pulmonary arterioles in each group showed that the RELM beta expression was obviously enhanced in COPD group compared with the control group, and the difference was statistically significant (P0.05); P H group showed strong positive expression, compared with group COPD (P0.05).3, Western blot detected RELM beta in the lung tissue of the patients, PI3K/Akt/m TOR signaling pathway protein expression Western blot results showed that the expression of Relm beta in the group was significantly higher than that of the control group, and the result was statistically significant. In group COPD, the results were statistically significant (P0.05); the expression of p-PI3K in group COPD was significantly higher than that in the control group (P0.05); the expression of p-PI3K in the PH group was significantly higher than that in the group COPD (P0.05), and the expression of p-Akt308 in the COPD group was significantly higher than that in the control group, and the results were statistically significant (P0.05). The expression of the group was significantly higher than that of the COPD group (P0.05), and the expression of p-Akt473 in the group COPD was significantly higher than that in the control group (P0.05), and the expression of p-Akt473 in the PH group was significantly higher than that of the COPD group, and the results were statistically significant (P0.05); P-M TOR in the COPD group was significantly higher than that of the control group, and the results were statistically significant. The expression of P-M TOR in the PH group was significantly higher than that in the COPD group, and the results were statistically significant (P0.05). The linear correlation analysis showed that the Relm beta protein in the lung tissue was positively correlated with WT%, WA%, PASP, and was negatively correlated with LA%, arterial oxygen pressure (Pa). The expression of Relm beta was statistically significant compared with that of the control group (P0.05). The plasma Relm beta expression in the PH group was significantly different from that in the COPD group (P0.05). The plasma HIF-1a expression in the COPD group was statistically significant compared with the control group (P0.05), and the HIF-1a expression in the plasma of the COPD group was compared with the PH group. The difference was statistically significant (P0.05). The linear correlation analysis showed that the expression of RELM beta protein in plasma was positively correlated with PASP, HIF-1a, and negatively correlated with Pa O2, but the expression of HIF-1a protein in the plasma was positively correlated with PASP, and negative correlation with Pa O2. The PI3K/Akt/m TOR signaling pathway is activated to participate in pulmonary vascular remodeling, resulting in the formation of PH.2. The expression of Relm beta and HIF-1a in the plasma of COPD related PH patients is significantly higher, and is positively correlated with the pulmonary arterial pressure.
【学位授予单位】：南华大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R544.1

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