Association Of Protein Z Promoter A-13G And Intron FG79A Polymorphisms With Coronary Artery Disease In Han Ethnic

Background and PurposeThe development of Coronary artery disease (CAD) is closely associated with the coagulation system. The disorders of coagulation and inhibition of fibrinolytic activity are often found in CAD patients. Coagulation factors play an important role in the development of atherosclerosis. The rupture of atheromatous plaque and thrombogenesis are the major pathophysiologic changes in acute myocardial infarction and unstable angina.Protein Z, a vitamin K-dependent glycoprotein, is a newly discovered coagulation factor which plays an important role in inhibiting coagulation in vivo and vitro. Human Protein Z plasma levels vary widelyin normal individuals. That may related to heredity such as its gene and gene polymorphisms. During the last few years, scanty and controversial data have published concerning the relationships between protein Z plasma concentration and atherothrombotic diseases, and between protein Z gene polymorphisms and atherothrombotic diseases. These data had attracted great interest recently.Studies showed recently that the gene of human protein Z was localized to chromosome 13 at band q34 and 14 single nucleio polymorphisms including protein Z intron F G79A and promoter A-13G polymorphisms were identified. No associations of the common polymorphisms were identified with vein thrombosis, and the relationships of these polymorphisms and artherothrombotic deserved further study. Lichy and his colleagues studied the association between protein Z intron F G79A, promoter A-13G polymorphisms and ischemic stroke. They found the A allele of protein Z intron F G79A polymorphism was a protective gene for early ischemic stroke. Santacroce R and his colleagues reported that the frequency of A allele in intron F was related to decrease of protein Z levels. The roles of the protein Z gene polymorphisms for the development and progression of coronary artery disease especially in Asian including Han ethnic group have not been reported. We investigated the protein Z intron F G79A and promoter A-13G polymorphisms by polymerase chain reaction and restrictionfragment length polymorphism (PCR-RFLP) in order to know the distribution of protein Z intron F G79A and promoter A-13G polymorphisms in Han ethnic group and the association with CAD. Meterials and Methods1. Subjects173 patients (127 males and 46 famales, the average age was 63.93±9.20 years) from the Cardiovascular Department of the Second Affiliated Hospital, Medical College of Zhejiang University were enrolled in the study who are Han ethnic without blood relationship. The selected coronary angiography was performed in all of the patients and more than one major coronary vessel with at least 50% stenosis were defined as CAD. The CAD group was divided into two subgroups: acute coronary syndrome subgroup (ACS subgroup) and non-acute coronary syndrome subgroup (Non-ACS subgroup). The ACS subgroup consisted of 47 unstable anginas and 27 acute myocardial infactions. And the Non-ACS subgroup consisted of 99 stable anginas. The control group consisted of 160 subjects (97males and 63 females, the average age was 58.12±11.67 years) with normal angiography.2. Methods2.1 Collection of clinical features: All subjects were enquired in detail for the histories of hypertension, diabetes, hyperlipidemia and smoking. The statures weight and major biochemical data were also measured.2.2 DNA was isolated from leukocytes using the method of salting out, and stored at-20℃.2.3 For analysis of the PZ gene polymorphisms of promoter A-13G and intron F G79A, a 272-bp fragment from the promoter region and a 320bp fragment from intron F region were amplified by polymerease chain reaction (PCR) using the PCR amplificon (MyCycler, BIO-RAD). Each PCR product was applied to a 2% agarose gel after ethidium bromide staining and analyzed with the imaging system.2.4 Analysis of polymorphisms of intron FG79A and promoter A-13G by restriction fragment length polymorphism (RFLP): The homozygous genotype with the restriction site was digested complet...