The Clinical And Experimental Study On Coronary Heart Disease Correlating To Adiponectin And Its SNP

This study consists of two parts,the first one is the clinical study about the relationship between SNP of adiponectin and coronary heart disease.The second part is the basic study about the mechanism for the anti-atherosclerosis role of adiponectin.Coronary heart disease is a multiple factor disease.The risk factors are age,smoking,hypertension,diabetes and genetic factor. Some studies showed genetic factor contribute to the development of cardiovascular disease .Especially, more people have interesting in the relationship between gene polymorphism and coronary heart disease is the hot study in recent years. At present many study have reported about the relationship between some adiponectin gene polymorphism sites and diabetes,insulin,metabolic syndrome and coronary heart disease .But there is no reporte about polymorphism of adiponectin which is associated with coronary heart disease In Chinese Han people.Atherosclerosis is a chronic inflammation including inflammatory cell infiltrate ,smooth muscle cell multiplication ,extracellular matrix increasing and thrombosis. In the early stages of atherosclerosis, circulating monocytes adhere to activated endothelial cells. Subsequently, monocytes differentiate into macrophages, leading to the accumulation of lipid rich foam cells by an uptake of modified lipoproteins.Chemotatic factors play an important role in the Atherosclerosis, monocytes adhering to endothelial cells are recruited by the Chemotatic factors. The express level of MCP-1, MIP-1αis more than the other Chemotatic factor in atherosclerotic disease.Adiponectin is an adipocytederived protein that is abundantly present in plasma .Adiponectin can inhibit atherosclerosis through many mechanisms of action,such as :1) Adiponectin improves endothelial function through stimulating AMPK signaling.2)Adiponectin inhibits monocytes adhesion , granulocytic differentiation and the produce of macrophage cytokine, 3) Adiponectin reduces expression of adhesion molecules in endothelial cells induced by TNF-αsuch as VCAM-1,endothelium-selectin and ICAM-1, 4) Adiponectin reduces the proliferation and ation of smooth muscle cells induced by platelet-derived growth factor in smooth muscle cells. 5) Adiponectin decrease the peroxide release which is induced by low density lipoprotein and ox-low density lipoprotein.At present some study have reported about the anti-inflammatory and anti-atherosclerosis mechanism action of adiponectin ,but we have not seen the study about the association of adiponectin and chemotatic factors.The first is the clinical study about the relationship between SNP of adiponectin and coronary heart disease. All participating individuals including 182 with normal and 176 with coronary heart disease are from the unrelated Chinese Han people of Jilin region.The polymerase chain reaction and restriction fragment length polymorphism ( PCR-RFL P) were performed to identify the polymorphism of exon 2 of adiponectin gene,and PCR direct sequencing were performed to identify the polymorphism of exon 3.The result shows the GG genotypic frequency of adiponectin gene SNP+45T/G in CHD patients is more than that in control subjects(P < 0.05). Adjusted for the presence of risk factors,such as age,sex,lipid,blood-fat,SBP, we can see GG and TG genotypic frequencies of adiponectin gene SNP+45 T/G in CHD patients were more than that in control subjects by the logistic regression analysis , compared with TT subject,TG subject:OR1:2.378,95%CI1:1.065-5.311, P1<0.05.CC subject:OR1:1.976,95%CI1:1.081-5.514, P1<0.05.。Statistical difference was also found in allele frequency compared between the two groups (P<0.05). We can see the patients carrying C allele can increase the risk of coronary heart disease.The result shows the CC genotypic frequency of adiponectin gene SNPY111H in CHD patients is more than the frequency in control subjects(P < 0.05). Adjusted for the presence of risk factors,such as age,sex,lipid,blood-fat,SBP, we can see CC and TC genotypic frequencies of adiponectin gene SNP Y111H in CHD patients is significantly higher than the frequencies in control subjects by the logistic regression analysis , compared with TT subject,TCsubject:OR1:1.185,95%CI1:0.97-1.44, P1<0.05;CC subject:OR1:2.22,95%CI1:1.16-4.27, P1<0.05。Statistical difference was also found in allele frequency compared between the two groups (P<0.05). that in CHD patients.The results shows The CC and TC genotypic frequencies of adiponectin gene SNP I164T in CHD patients were more than the frequencies in control subjects(P < 0.05). adjusted for the presence of risk factors,such as age,sex,lipid,blood-fat,SBP, we can see CC and TC genotypic frequencies of adiponectin gene SNP I164T in CHD patients was significantly higher than the frequencies in control subjects by the logistic regression analysis (P1<0.05).Statistical difference was also found in allele frequency compared between the two groups (P<0.05).In our study ,by analysing the relation between coronary heart disease subgroup and SNP+45T/G of adiponectin, no significant associations were observed between SNP+45T/G with clinical phenotype and severity of coronary arteries.On the other hand we detected the serum lipid levels of CHD patients and control patients.We can see the level of TC, TG,LDL-C in CHD patient were higher than that of the control patients,and HDL-C was significant lower in CHD patient than that of the control patients. The serum lipid levels of CHD patients carrying SNP+45 T/G have significant difference distributed in different genetype,the level of CHOL-C and LDL of subjects carrying G allele was significantly higher than that of subjects carrying T allele,meanwhile the level of HDL-C was lower.Both in acute coronary syndrome group and stable angina group ,T/T genotype was the most frequency one of SNPI164T ,but the frenquency of C/C and TC genotype in acute coronary syndrome group was significantly higher than that in stabilizing angina group(P<0.05). Statistical difference was also found in allele frequency compared between the two groups (P<0.05).The SNPI164T of adiponectin is accociated with the development from stabilizing angina to acute coronary syndrome .The study on relationships among SNPY111H,SNPI164T of adiponectin and serum lipid levels show that no significant difference distributed in difference genetype of SNPY111H.The serum lipid levels of CHD patients carrying SNPI164T have significant difference distributed in difference genetype,the level of LDL-C of subjects carrying C allele was significantly higher than that of subjects carrying T allele,meanwhile the level of HDL-C was decreased(P<0.05). According No SNP R221S ,SNP H241P and new SNP were detecedt in exon 3 , there were no SNP R221S ,SNP H241P in Chinese Han people of jilin region .The second part of study is to observe the influence of adiponectin on TNF-α-induced expression of MCP and MIP-1αin human umbilical vein endothelial cells(HUVECs). The expression of MCP and MIP-1αin protein level and mRNA level was deteced with experimental method of cell immunochemistry and reverse transcription polymerase chain reaction( RT-PCR).Through cell immunohemistry method ,we can see TNF-αmay induced the protein expression of MCP-1 and MIP-1αin HUVECs. Adiponectin of different density all can inhibit the expression of MCP and MIP-1αand the inhibitory actions were concentration-dependent .The result showed more than moderately density adiponectin can decrease the level of protein expression of MCP-1 and MIP-1α,and Statistical difference was found in the level of protein expression of MCP and MIP-1αcompared with subject only with TNF-αstimulated and the low dnsity adiponectin subject.Through RT-PCR expriment,we can see adiponectin of different density all can inhibit the level of mRNA expression of MCP-1 and MIP-1αand the inhibitory efects were concentration-dependent. More than moderately density adiponectin can decrease the level of mRNA expression of MCP-1 and MIP-1α,and Statistical difference was found in the level of mRNA expression of MCP-1 and MIP-1αcompared with subject only with TNF-αstimulate and the low dnsity adiponectin subject. All the result showed that more than moderately density adiponectin can decrease expression of MCP-1 and MIP-1αin the endothelial cell which can decrease the monocyte tendency to endothelial,and play the anti-atherosclerotic role.The conclusion of our study is single nucleotide polymorphism of adiponectin such as SNP +45T/G,SNPY111H,SNPI164T is associated with the coronary heart disease .The SNP +45T/G and SNPI164T of adiponectin are associated with the dyslipidmia. The SNPI164T of adiponectin is associated with the clinical phenotype of CHD and severity of coronary arteries. More than moderately density adiponectin can decrease the expression of MCP-1 and MIP-1αin the endothelial cell . Our study may provid the new theory for the anti-atherosclerosis role of adiponectin,and the new treatment methods of atherosclerosis at clinic medical science .