The Research Of Technology For Vascular Endothelial Cells Damage In Acute-phase Patients With Coronary Heart Disease

Objective To investigate the alteration of vascular endothelial cells(VECs) and circulation endothelial cells(CECs) damage in acute-phase Patients with Coronary Heart DiseasConstruct technology platform of vascular endothelial cells damage, in order to provide new theory evidence and laboratory indicatrix of earlier-phase prophylaxis and diagnosis for nosogenesis,development,turnover of Coronary Heart DiseaseMethods 65 acute-phase Patients with Coronary Heart Disease of preparing for operation of heart pass-b were chosen from chest surgery in the second hospital of lanzhou university; and 38 health adults were controlled, they were from medical examination center in the second hospital and first hospital of lanzhou university. Number of the CECs was counted by flow cytometry(FCM), the damage of nuclear DNA for CECs was assayed by single cell gel electrophoresis (SCGE) and agarose gel electrophoresis(AGE) in 65 acute-phase patients with Coronary Heart Disease. Damage of VEC ultrastructure were investigated by electronmicroscope.Results Number of the CECs was higher in patients with CHD than health group (32.6±9.1 /ml vs 4.17±1.23/mL, P<0.01), Significant correlation was calculated between the damage of VEC and the number of CEC(r=2.83,P<0.05). The nuclear DNA of Acute Coronary Heart Disease group was obviously demonstrated like comet and majority of nuclear DNA were demonstrated like circle in control group. The rate of comet cell was investigated and results revealed that Acute Coronary Heart Disease group had a higher percentage of comet cells than control group (92.92%±6.95% and 10.49%±4.45%, respectively, P<0.001), and their tail DNA content was significantly increased in patients with Coronary Heart Disease (41.12%±3.77% and 7.33%±2.53%, respectively, P<0.001 ). The multi-factors related analysis showed that damage of VEC had significant correlation with and the nuclear DNA damage in patients with Acute Coronary Heart Disease (r=3.43,P<0.05). simultaneously, the irregular shape was shown in Coronary Heart Disease group by AGE, and the shape of control group was uniform single. Electronmicroscope shows : cellular membrane of VEC was obviously damaged, exfoliation, smooth muscle was exposed.; chondriosome and golgiosome were obviously engorged, cytolysosome was increased and the last manifestation was necrosis in patients with Acute Coronary Heart Disease as CEC.Conclutions 1. CEC can be extracted from the blood of patients with Acute Coronary Heart Disease by means of density gradient centrifugation of Percoll in this study, and CEC can be numerated by FCM.2. Significant correlation was calculated between the damage of VEC and the number of CEC. It is demonstrated that CEC was derived from VEC.3. SCGE shown that DNA of CEC was damaged obviously in patients with Acute Coronary Heart Disease. The cell membrane of VEC was damaged, and the mitochondrion, Golgi body and endoplasmic reticulum were swelled obviously. Lysosome increased largely and some cells were becoming dissolved.4. Both the number of CEC and the SCGE were the important methods to reflect the damage of VEC. The application of number of CEC with FCM was better in early stage than other stages in patients with Acute Coronary Heart Disease. When disease become worse, the number of CEC can not reflect the damage of VEC, the defect can be compensated by undergoing the single cell gel electrophoresis. In addition, in the library without FCM, SCGE is also useful experimental method to detect the damage of DNA.