慢性阻塞性肺疾病患者肺组织基质金属蛋白酶2与细胞黏附因子的表达

Expressions of intercellular adhesion molecule and matrix metalloproteinase 2 in lung tissues of patients with chronic obstructive pulmonary diseases

目的:研究慢性阻塞性肺疾病患者肺组织基质金属蛋白酶2、细胞黏附因子蛋白及mRNA在肺内的分布和表达、相互作用及与气流阻塞的关系。方法:收集2005-03/09哈尔滨医科大学附属第一医院和附属第三医院胸外科手术肺肿瘤患者边缘肺组织(距肿瘤组织>5cm)标本28份。根据吸烟史、有无慢性阻塞性肺疾患病史、胸部X射线片、肺功能将标本来源患者分为两组,对照组9例,无吸烟史也无慢性阻塞性肺疾病;慢性阻塞性肺疾患组19例,吸烟伴有慢性阻塞性肺疾病。通过肺功能检测两组患者第一秒用力呼气容积百分比、用力肺活量百分比、第一秒用力呼气容积/用力肺活量百分比肺功能变化指标。采用免疫组织化学方法和反转录-聚合酶链反应法检测人肺组织基质金属蛋白酶2、细胞黏附因子蛋白和mRNA的表达,并进行相关性分析。结果:纳入患者28例,均进入结果分析。①肺功能变化指标:两组比较第一秒用力呼气容积百分比、用力肺活量百分比、FEV1/FVC/%差异显著(P<0.01)。②免疫组织化学染色结果:基质金属蛋白酶2高表达于慢性阻塞性肺疾患组肺泡上皮细胞、支气管上皮细胞、血管平滑肌细胞、肺泡巨噬细胞、间质细胞(积分值为2.47±0.70)。对照组表达非常弱(积分值为1.22±0.44),两组比较差异显著(P<0.01);细胞黏附因子高表达于慢性阻塞性肺疾患组肺泡上皮细胞(积分值2.58±0.51),对照组表达非常弱(积分值0.67±0.87),两组比较差异显著(P<0.01)。③反转录-聚合酶链反应结果:基质金属蛋白酶2平均吸光度值(A)半定量分析,慢性阻塞性肺疾患组高于对照组,差异显著(0.54±0.14,0.19±0.08,P<0.01)。细胞黏附因子平均吸光度值(A)半定量分析,慢性阻塞性肺疾患组高于对照组,差异显著(0.62±0.15,0.37±0.11,P<0.01)。④相关分析:基质金属蛋白酶2mRNA表达及免疫组织化学表达与第一秒用力呼气容积百分比、第一秒用力呼气容积/用力肺活量百分比呈直线负相关(r=-0.577,-0.768,-0.531,-0.591熏P<0.05);肺组织细胞黏附因子mRNA及免疫组织化学与第一秒用力呼气容积百分比、FEV1/FVC/%呈直线负相关穴r=-0.584,-0.529,-0.580,-0.721,P<0.01雪。慢性阻塞性肺疾患患者肺组织基质金属蛋白酶2、细胞黏附因子mRNA呈直线正相关穴r=0.535,P<0.01雪,慢性阻塞性肺疾患患者肺组织基质金属蛋白酶2、细胞黏附因子免疫组织化学分析呈直线正相关穴r=0.664熏P<0.01雪。结论:慢性阻塞性肺疾患患者肺组织基质金属蛋白酶2增多使气道结构细胞失去正常的支持作用,通过细胞黏附因子促进炎症细胞的趋化、移行,聚集于血管壁,释放炎性因子,迁移进入细胞外基底膜及气道上皮细胞,参与气道壁的炎症反应,引起肺组织的重构,引起及加重慢性阻塞性肺疾患患者的气流阻塞。

AIM： To evaluate the distribution, expression and interaction of matrix metalloproteinase 2（MMP-2）, intercellular adhesion molecule-1 （ICAM-1） protein and mRNA in the lung tissues of patients with chronic obstructive pulmonary diseases （COPD） and its correlation with airflow obstruction. METHODS： Totally 28 edge lung tissue samples （the distance of tumor tissue 〉5 cm） were selected from the chest surgical excisions of lung tumor patients at the First Hospital and Third Hospital, Affiliated to Harbin Medical University. According to the smoking history, COpD history, X ray sheet of chest and pulmonary function, all the patients were divided into two groups： Control group （n=9 without smoke or COPD history） and COPD group （n=19 with smoking and COPD history）. The forced expiratory volume in first second （FEV1） percentage, forced vital capacity （FVC） percentage and FEV1/FVC percentage were detected and taken as the pulmonary function change index. The expressions of MMP-2, ICAM-1 protein and mRNA in lung tissues were detected by immunohistochemistry method and reverse transcription-polymerase chain reaction （RT-PCR）, and the correlative analysis was conducted. RESULTS： Totally 28 patients were involved in the result analysis.①The change index of pulmonary function： Difference was significant between control group and COPD group in FEV1 percentage, FVC percentage, and FEV1/FVC percentage （P 〈 0.01）. ②Immunohistochemistry staining result： MMP-2 was highly expressed in alveolar epithelial cells, bronchial epithelial cells, vascular smooth muscle cells, alveolar macrophages and interstitial cells in COPD group, compared with the control group[（2.47±0.70）, （1.22±0.44）], with the significant difference（P 〈 0.01）; The expression of ICAM-1 was high in alveolar epithelial cells in COPD group, compared with the control group [（2.58±0.51）,（0.67±0.87）], with the significant differenee（P 〈 0.01）.③RT-PCR result： The average absorbance （A） value of MMP-2 semi-quantitative analysis was higher in CDPD group than in the control group, with the significant difference [（0.54±0.14）,（0.19±0.08）, P 〈 0.01].Similarly, ICAM-1 expression was significantly higher in the COPD group compared with controls[（0.62±0.15）,（0.37±0.11）, P 〈 0.01]. ④The correlative analysis： Both mRNA and protein levels of MMP-2 were inversely correlated with FEV1 percentage and FEVI/FVC percentage respectively （r=-0.577,-0.768,-0.531,-0.591,P 〈 0.05）; Similarly, inversely correlation was also found between ICAM-1 expression （mRNA and protein） and FEV1 percentage and FEV1/FVC percentage （r=-0.584, -0.529,-0.580,-0.721,P 〈 0.01）. The expression of MMP-2 and ICAM-1 was positively correlated beth on mRNA （r=0.535, P 〈 0.01） and protein levels （r=0.664, P 〈 0.01）. CONCLUSION： Over-expressions of MMP-2 in lung tissues of patients with CDPD may lead to the abnormal support action of airway structural cells, and improve the chemotaxis and migration of inflammatory cells through ICAM-1, release certain inflammatory factors, move into the basement membrane and airway epithelial cells. In addition, highly expressed MMP-2 may participate in the inflammatory reaction of airway wall and result in the reconstitution of the lung tissues, which causes and aggravates the airway obstruction in patients with CDPD.