Study On The Correlation Between Acylation Stimulating Protein,Complement C3 And Atherosclerosis

Background:Atherosclerosis (AS) is the important pathogenesis of cardiovascular and cerebrovascular disease,and its formation is a complex pathological process. The pathogenesis of atherosclerosis involves a wide variety of pathological reactions, such as endothelial cell damage, lipid infiltration,inflammatory cell infiltration,plaque rupture, thrombogenesis and so forth.Of note,in the development of atherosclerosis, acylation stimulating protein(ASP) plays a pivotal role. ASP is a kind of small molecular protein which had been isolated from human plasma by Cianflone in 1989. It is a kind of autocrine hormone by adipocyte. C3 is precursor protein of ASP and affects the produce of ASP. C5L2 is the only functional receptor of ASP, while,ASP biological function is closely related to the C5L2 receptor. The current research was mainly focused on whether ASP regulated glucose and lipid metabolism.It has been documented that ASP and C3 are closely related with obesity and type 2 diabetes. Metabolic syndrome(MS) and coronary heart disease (CHD) are the common diseases in clinic.Atherosclerosis is likely to occur in MS and coronary heart disease (CHD).Therefore, in the part I, we research the correlation between ASP, C3 and ABI,baPWV, CIMT in order to discuss whether they participate in the occurrence and development of atherosclerosis;In the part Ⅱ,we research the correlation between ASP, C3, ASP/C5L2 gene polymorphism and coronary heart disease in order to discuss whether they involve in the occurrence and development of coronary heart disease. This will be a new thinking for early detection and intervention for arterosclerosis.Part Ⅰ The relevant research of acylation stimulating protein and complement C3 and metabolic syndrome atherosclerosisObjectiveBy measuring the level of ASP, C3 in MS group and control group and by analyzing the correlation of ASP, C3and baPWV, ABI, CIMT,we explore the relationship between the ASP, C3 and atherosclerosis.MethodsA total of 76 patients,who were diagnosed with MS,were included as the MS group,while, the other 42 voluteers,who were excluded hypertension,diabetes mellitus,abdominal obesity and dyslipidemia and so on,were taken as the control group. Two groups were all subjected to the history collection and physical examination, Enzyme-linked immunosorbent assay was used to measure ASP concentration. Immunoturbidimetric assay was employed for the determination of complement C3 concentration. FBG,TC, TG,HDL-C,LDL-C were measured by clinical laboratory. Carotid ultrasound analysis was performed for carotid intima-media thickness measurement and Omron arteriosclerosis detector was used for brachial-ankle pulse wave velocity and ankle-brachial index measurements. Statistical analyses were performed using the Statistical Package for Social Sciences software (SPSS version 13.0, USA). Measurement data were expressed as means± SD. Comparison between two groups was performed using t-test. Correlations among variables were based on analysis of Pearson’S correlation and partial regression.Multivariate logisitic regression analysis was used to analyze the correlations between MS and all other parameters. P<0.05 was considered statistically significant.Results(1) The age and gender compositions were no statistically differences between MS group and control group (P>0.05).The waist circumference, BMI, SBP, DBP, FBG, TG, TC, LDL-C were higher in MS group than those in control group(P <0.05).(2) The baPWV, ABI and CIMT were differences between MS group and control group.(3) The serum ASP and C3 concentration were no significant difference between MS group and control group. Multiple factor regression analysis showed that ASP, C3 and MS had no significant correlation (P> 0.05).(4) The serum ASP, C3, SBP, FBG, LDL-C and HDL-C concentrations were differences between in MS+AS group and MS-AS group (P<0.05). SBP,FBG were higher in abnormal baPWV group than in normal baPWV group.FBG,LDL were higher in abnormal ABI group than in normal ABI group.But ASP and C3 were no significant difference in normal and abnormal groups of baPWV,ABI.(5) The bivariate correlation analysis showed that serum ASP,C3 level and waist circumference,BMI, FBG, TG had positive correlations(P<0.05),whereas had no significant correlation with baPVWV,ABI (P>0.05).After controlling metabolic factors,the correlation between ASP, C3 and CIMT was remained(P <0.05).ConclusionASP and C3 are associated with CIMT, and they may play a role in the pathogenesis of atherosclerosis.Part II The relevant research of acylation stimulating protein, C3 and ASP 301T>C,C5L2 698C>T gene polymorphism and coronary heart disease (CHD)Objective(1) By measuring the serum levels of ASP, C3 in CHD group, we investagated the relationship between ASP, C3 and coronary heart disease and the severity of coronary.(2) By testing ASP 301 T>C and receptor C5L2 gene 698C>T polymorphism in Shandong Jinan regions and different ethnic CHD patients, we explored the relationship with coronary heart disease (CHD).MethodsA total of 450 patients diagnosed CHD were CHD group, (245 Han individuals population and 205 Hui individuals).500 volunteers were taken as the control group, han (250 population and 250 hui population). All subjects had no genetic connection. FBG,TC, TG,HDL-C,LDL-C were measured by clinical laboratory. DNA was extracted using TIANGEN RelaxGene Blood DNA samples System.The genotypings of SNP were determined using the TaqMan probe technique.In addition, we selected 62 patients who had been determined by coronary angiography and we selected 42 volunteers from control group. They were all conducted the history collection and physical examination, Enzyme-linked immunosorbent assay was used to measure ASP concentration. Immunoturbidimetric assay was employed for the determination of complement C3 concentration.Western blot was used to determine the protein expression levels of C3. Quantitative RT-PCR analysis was used to determine the serum mRNA expression levels of C3.Statistical analyses were performed using the Statistical Package for Social Sciences software (SPSS version 13.0, USA). The Hardy-Weinberg equilibrium was assessed by x2 test. Continuous variables were compared using the t-test and analysis of variance. Multivariate logisitic regression analysis was used to analyze the correlations between CHD and all other parameters.χ2 analysis were used for the comparison of allele and genotype frequencies.Analysis of covariance was used for the comparison of blood glucose and lipid levels between different genotypes.P<0.05 was considered statistically significant.Results(1) The age and gender compositions were no statistically significant differences between CHD group and control group(P>0.05). SBP,DBP,FBG,TG, TC, LDL-C were different between CHD group and control group(P<0.05).(2) The concentrations of ASP,C3 in CHD group were obviously higher than, those in control group(P<0.01).(3) The concentrations of ASP,C3 in the moderate and severe coronary lesion groups were obviously higher than those in none and mild lesion groups (P< 0.05). Bivariate correlation analysis showed that the serum ASP, C3 level had clear correlation with coronary Gensini score (P< 0.05).(4) The protein expression and mRNA expression levels of C3 in the moderate and severe coronary lesion groups were higher than those in no and mild lesion groups (P< 0.05).(5) Multivariate logisitic regression analysis showed that ASP and C3 were correlated with CHD (P< 0.05).(6) Hardy-Weinberg equilibrium test:χ2 values of ASP genotype distribution in the group of Han and Hui population were 0.875 (P=0.38) and 0.087 (P= 0.75), and conform to the Hardy Weinberg equilibrium.They had group representative; x2 value of C5L2 site genotype distribution in the group of han and hui population were 0.863 (P=0.37) and 1.439 (P=0.26), and conform to the Hardy Weinberg equilibrium.They had group representative.(7) The frequency of the allele genes T,C and the genotype of ASP had no significant difference in the CHD group and the control group in han subjects,but they had difference in Hui subjects.The frequency of homozygote carriers of the CC genotype of ASP was significantly higher in CHD group than in control group in Hui subjects. The frequency of the allele genes C,T and the genotype of C5L2 had difference in the CHD group and the control group both in han and hui subjects. The frequency of the heterozygote carriers of the CT genotype of C5L2 was significantly higher in CHD group than in control group.(8) There were significant correlation between the genotype of ASP 301 CC genotype and C5L2 698 CT genotype and the blood glucose in han and hui populations,but the correlation with the blood lipid were different. There were correlations only between the genotype of ASP 301 CC genotype and C5L2 698 CT genotype and TG in hui populations.ConclusionASP and C3 may play a role in the pathogenesis of coronary heart disease (CHD). Receptor C5L2 698 C>T may be the susceptibility gene for coronary heart disease (CHD) in Han and Hui populations in shandong jinan district, the ASP 301 T>C may be the susceptibility gene for coronary heart disease only in shandong jinan district hui population.