The Effect Of Hp Plasma Concentration And Polymorphism On The CHD Patient

BackgroundCoronary heart disease is a multi-factors disease,many studies have proved that inflammation play a significant role in the process of its progress.Serum concentration of Acute-phase-reactants can disclose the severity of the CHD.Hp has a function of anti-inflammation,and different genotype of Hp has different ability of anti-inflammaiton, especially in diabetic patient.Acute-phase-reactants can be used as biomarker of CHD, The biomarkers may be helpful to assess the CHD. We hypothesize that these diabetic patients with Hp2-2 genotype should be paid more attention on the complication of cardiovascular, because they have high level of oxidation.Objectives:1. In order to explore the different plasma level of lipoprotein and acute phase proteins in the CHD group and control group. Access the relationship between the extent of coronary artery disease and protein blood concentration.2. To find out weather Hp phenotype has an effect on the pathogenesis of CHD, Whether Diabetic patients have an susceptibility of suffer from cardiovascular complication.Study populationsBoth the CHD and healthy populations were obtained from patients in FUWAI Hospital during August 2008 to December 2009. Inclusion criteria of the studies: 1.CHD group, Coronary angiography was examined and at least one coronary artery was blocked more than 50%; 2.Control group, coronary angiography was normal, and other cardiovascular examination including Holter, Ultrasound and so on can exclude the other cardiovascular disease.3 Diabetic patients with coronary heart disease, symptom of Diabetes, the blood sugar high than 11. lmmol/L at any time or high than 7.0mmol/L with a empty stomach,or high than 11.1mmol/L after taking 75g sugar 2 hours in OGTT test. At least one coronary artery was blocked more than 50%. The study was improved by the Independent ethics committee of Fuwai Hospital. Written informed consent was obtained from each participant before the study.MethodsBlood samplingPeripheral venous blood samples were collected in the morning after an overnight fast. Routine blood biochemistry was measured. Serum concentrations of total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C) and triglycerides (TG),LP(a),hs-CRP were measured using a Beckman Synchrom CX5 (Fullerton, California,USA). Concentration of Hp was measured by Beckman Coulter,Inc (American Beckman company). Each EDTA-anticoagulated blood sample(4ml) was centrifuged at 4℃(3000rpm for 10 minutes) and the supernatant was collected for the test of Hp plasma concentration and Hp genotype.Hp phenotypes native polyacrylamide gel electrophoresisHp phenotypes of individuals were determined by Tetramethyl-benzidine/hydrogen peroxide staining after native polyacrylamide gel electrophoresis of Hphemoglobin (Hb) complexes. Briefly, the Hp in serum was converted into Hp-Hb complex by mixing 15μL of serum with excess Hb, and electrophoresed on 7%native polyacrylamide gel and with a initial voltage of 80 V and increased up to 120 V when the dye front reached the separating gel. After electrophoresis, the gel was soaked in 1%benzidine/20%acetic acid solution/5%DMSO, and a small amount of hydrogen peroxide was immediately added to it while shaking. The bands representing Hp-Hb complexes in the gel could be visualized by the peroxidase activity of Hb. In the resultant electrophoretic pattern, Hp 1-1 type shows only a single band, whereas Hp 2-2 type shows slow-migrating polymer bands. Hp 2-1 type represents both 1-1 type and polymer forms.Coronary angiography integrated systemThe Gensini score assesses the severity of coronary artery disease:it grades narrowing of the lumen of the coronary artery and scores it as 1 for 1-25%narrowing, 2 for 26-50%narrowing,4 for 51-75%,8 for 76-90%,16 for 91-99%and 32 for a completely occluded artery. This score is then multiplied by a factor according to the importance of the coronary artery. The multiplication factor for a LM lesion is 5, it is 2.5 for proximal LAD and LCX lesions,1.5 for a mid-LAD lesion, and 1 for distal LAD, mid/distal LCX and right coronary artery lesions. The multiplication factor for any other branch is 0.5.Results1. Plasma level of HDL-C, CRP, LP (a) and Hp have a significant differences between the control group and the CHD group, but there is no significant difference in the plasma level of TC, TG, LDL-C.2. The rate of Diabetes, Hypertension, Family cardiovascular history and smoking are high in CHD group compare with control group, and this difference have statistically significant.3. The plasma concentration of HDL-C,CRP,LP(a),TC,TG,LDL-C have no significant differences among the different sub-group of CHD group according the Gensini score.4. There are no statistically significant in the plasma concentration of TC, TG, LDL-C HDL-C,LP(a),Hp,CRP between the CHD and DB with CHD group.5. There is no significant difference of Hp Gene frequencies among the control group, CHD group and DB with CHD group.6. In the DB with CHD group, the Gensini integrate score are much high than CHD group.7. Both in CHD and DB* CHD group, the Hp2-2 sub-group have high Gensini integrate score.Conclusion1. In Chinese population, the Diabetes, Hypertension, Family history and smoking are all the risk factor of CHD, High plasma level of Hp,CRP,LP(a) also have relationship with the CHD just as the low plasma level of HDL-C.2. In this study, the plasma level of HDL-C, Hp, CRP, LP (a) can't assess the severity of CHD in Chinese population.3. The Hp genotype have no relationship with the susceptibility of CHD.4. Coronary heart disease symptom in Diabetic patients is more serious than CHD patients without Diabetes.5. The Hp genotype has relationship with the extent of coronary heart disease both in CHD and DB with CHD Chinese population in this small case study.