The Measure Of Rho-kinase Of Serum And Peripheral Blood Monocytes In Patients With PAH Realated COPD

Objective:To determinate ROCK1 of the serum and peripheral blood moncytes in healthy people,smokers and patients with PAH realated COPD.Methods:1.Objects and settingsTen patients in pulmonary arterial hypertension(PAH) associated with chronic obstructive pulmonay disease(COPD) who were hospitalized in the third hospital of Changsha from December 2010 to April 2011 were chosen to groupC (male 6, female 4, average age 65.4±8.0).Ten healthy people who take health exam at the same time were chosen to groupA (male 5, female 5, average age 62.5±7.0) and groupB(who have smoke habbits) (male 7, female 3, average age 63.7±9.0). All patients in groupA were diagnosed according to The COPD Guideline Published by Chinese Medical Committee in 2007 and the Guide-line of pulmonary hypertension by ESC in 2009.2.Pulmonary function test:All objects take on pulmonary function test included forced vital capacity(FVC),forced expiratory volume at first minute(FEV1)and FEV1/FVC,the percentage of predict FEV1 (FEV1%). 3.Pulmonary arterial systolic pressure testAll objects accepted pulmonary arterial systolic pressure test by echocardiography.4.The levels of ROCK1 in serum test1)Sample collection:3 milliliter blood was collected from each objects and extracted the serum through centrifuge from each sample for 15 minutes with a speed of 3000r/min, temperature of 20℃.The serum were preserved for ELISA in a refrigerator with-80℃2)Test method:The levels of ROCK1 in serum of all objects were tested by enzyme linked immunosorbent assay(ELISA).5.The levels of ROCK1 in moncytes test1)Sample collection:20 milliliter blood was collected from each objects. The peripheral blood mononuclear cell(PBMC) were separated by percoll, and moncytes were adherenced. The cells were preserved for ELISA in a refrigerator with-80℃.2) Measure method:Splitting the cells with freeze-thawing technique,the levels of ROCK1 in supernatant in all objects were measured by enzyme linked immunosorbent assay(ELISA). Results:1.The pulmonary arterial systolic pressure(PASP) in group C were significantly higher than those of group A and group B(P<0.01).The pulmonary arterial systolic pressure(PASP) of group B were higher than those of group A(P<0.05).2.The levels of ROCK1 in serum of group C were higher than those of group A and group B(P<0.05). There was no difference between group B and group A(P>0.05).3. The levels of ROCK1 in moncytes of group C were higher than those of group A and group B(P<0.05). There was no difference between group B and group A(P>0.05).4.The cell court of moncytes in group C was more higher than those of group A and group B(P<0.05).5.The levels of ROCK 1 in serum had a positive relationship with pulmonary arterial systolic pressure(r=0.736,P<0.05);The levels of ROCK1 in moncytes had a positive relationship between pulmonary arterial systolic pressure(r=0.661,P<0.05).Conclusions:1. The levels of ROCK1 in serum and the moncytes increased with pulmonary arterial systolic pressure.It may be a marker of the severity of patients with PAH realated COPD.2. The levels of ROCK1 in serum and moncytes showed no significant difference between the healthy people and smokers.We need to prove the difference of Rho-kinase activity between them by large samples.3.Smoking maybe correlate with the pulmonary arterial systolic pressure up.We hope to prove the relationships between Smoking index and pulmonary arterial systolic pressure increased in future studies.