慢性阻塞性肺疾病相关性肺动脉高压患者外周血单核细胞Rho激酶水平测定

摘要

目的:通过比较Rho关联含卷曲螺旋蛋白激酶1(Rho-associated coiled-coil containing protein kinase 1,ROCK1)在不同研究对象外周血单核细胞中的表达规律,探讨RhoA/Rho激酶信号通路在慢性阻塞性肺疾病相关肺动脉高压中的作用.方法:选取2010年12月至2011年4月在长沙市第三医院体检的健康不吸烟者10人(A组),同期同院呼吸内科住院的慢性阻塞性肺疾病不合并肺动脉高压者10人(B组),以及同期同院呼吸内科住院的慢性阻塞性肺疾病相关性肺动脉高压患者10人(c组).分别抽取各研究对象静脉血20 mL,以percoll非连续密度梯度离心沉降法分离单个核细胞,并培养使单核细胞贴壁.刮下贴壁细胞裂解后取上清液,用ELISA法分别检测3组ROCK1水平.所有研究对象经肺功能仪检查肺功能,以彩色多普勒测定肺动脉收缩压.结果:1)3组研究对象中c组肺动脉收缩压显著高于A组与B组,(均P＜O.01).2)3组研究对象中c组外周血单核细胞ROCK1水平高于A、B两组(均P＜0.05)；B组外周血单核细胞ROCK1水平高于A组(P＜0.05).3)3组研究对象中外周血单核细胞中ROCK1水平与肺动脉收缩压呈正相关关系(r=0.661,P＜0.05).4)3组研究对象中外周血单核细胞中R.CK1水平与肺功能FEV1％无相关关系(r=0.131,P＞0.05).结论:Rho激酶在肺动脉高压发病机制中具有重要意义；单核细胞中ROCK1水平可能反映慢性阻塞性肺疾病相关肺动脉高压患者的病情严重程度.%Objective: To determine effects of the RhoA/Rho kinase signaling pathway on patients with pulmonary arterial hypertension related to chronic obstructive pulmonary diseases by testing levels of Rho-associated coiled-coil containing protein kinase 1(ROCK1) in peripheral blood monocytes in healthy subjects, patients with chronic obstructive pulmonary diseases (COPD), and patients with pulmonary arterial hypertension related to COPD.Methods: Ten healthy subjects (Group A), 10 patients with COPD (Group B), and 10 patients with pulmonary arterial hypertension related to COPD (Group C) were enrolled, all of whom were hospitalized in the Third Hospital of Changsha between Dec. 2010 and Apr. 2011. Twenty-milliliters of blood was collected from each subject. The peripheral blood mononuclear cells (PBMC) were separated by Percoll and, monocytes were incubated. Levels of ROCK1 in the three groups were measured by ELISA. The pulmonary function was measured by spirometric tests, and the pulmonary arterial systolic pressure (PASP) was detected by color Doppler echocardiogram. Results: 1)The PASP in Group C was significantly higher than that of Groups A and B(P<0.01). 2) The levels of ROCK1 in monocytes of Group C were higher than those of Groups A and B(P<0.05). The levels of ROCK1 in monocytes of Group B were higher than those of Group A (P<0.05). 3) The levels of ROCK1 in monocytes of the three groups showed a positive correlation with PASP(r=0.661, P<0.05). 4) The levels of ROCK1 in monocytes of the three groups showed a negative correlation with forced expiratory volume at the first second/ forced vital capacity (FEVl%)(r=0.131, P>0.05).Conclusion: Rho kinase plays a key role in the pathogenesis of pulmonary arterial hypertension. The ROCK1 may be a marker of the severity of pulmonary arterial hypertension related to COPD.