Clinical And Basic Study On The Relationship Between MPO/PON1 And HDL Function In Patients With Coronary Heart Disease

Objective:Analysis of the changes of myeloperoxidase（MPO）and paraoxonase-1（PON1）in plasma of patients with acute myocardial infarction（AMI）and non-coronar y heart disease（N-CHD）,and then cultivate THP-1-derived macrophages with different groups of serum,by measuring the rate of cholesterol efflux,ABCA1,ABCG1 protein and mRNA expression in order to show changes in the function of HDL-C,and determining the level of IL-6 and TNF-αserum to indicate the inflammatory state of macrophages,to investigate the relationship between MPO/PON1 and HDL in patients with coronary heart disease and the protective effect of quercetin on HDL function.Methods:1.Part of clinical research:（1）150 patients with AMI and N-CHD were selected to collect basic clinical data and biochemical indexes.（2）Fasting venous blood in the two groups were centrifuged to obtain serum and plasma,plasma levels of MPO and PON1were measured by ELISA.2.Part of vitro research:（1）Cultured human monocytes（THP-1）,and induced macrophages.（2）Respectively,from the AMI and N-CHD patients with apoB-deficiency serum intervention experiment,then divided into four groups:the experimental group（with AMI patients serum intervention）,the Que group（with AMI patients serum and quercetin intervention）,the control group（with N-CHD patients serum intervention）,blank group（without any intervention）,then cholesterol efflux rate were determined,reverse cholesterol transporters protein ABCA1 and ABCG1 protein and mRNA expression were measured by Western-blot and PCR,inflammatory cytokines TNF-αand IL-6 levels were measured by ELISA.Results:1.There were significant differences in age,sex,hypertension,diabetes mellitus,smoking and cardiovascular family history between AMI and the N-CHD group（P<0.01）,AMI group age was larger than N-CHD group,predominantly male gender.Although there was difference between the two groups about history of drinking,but the difference was not statistically significant.2.Compared two groups of blood indexes,TG,TC,LDL-C,apoB,hs-CRP in AMI group were higher than in N-CHD group,HDL-C,apoA,apoA/apoB were lower than N-CHD group,the difference was statistically significant（P<0.01）.Meanwhile,the difference of ABO blood type between the two groups was also statistically significant（P<0.01）.3.There was no linear correlation between hs-CRP and HDL in AMI group,but a negative correlation between hs-CRP and HDL in N-CHD group,and the correlation coefficient r=-0.791.4.AMI group plasma MPO levels were significantly higher than N-CHD group,PON1 activity was significantly lower than N-CHD group,the ratio of MPO/PON1 in AMI group was higher than N-CHD group,the difference was statistically significant（P<0.001）.5.There is no linear correlation between MPO/PON1 and HDL-C in AMI group.6.The difference of cholesterol efflux in the four groups was statistically significant（P<0.01）;compared with the control group,the cholesterol efflux rate decreased in the experimental group and the Que group（P<0.01）;it was significantly higher in the Que group compared with the experimental group（P<0.01）;compared with the blank group,it significantly increased in the experimental group and the Que group（P<0.01）.7.Compared with the control group,the expression of ABCA1 and ABCG1 protein decreased in experimental group and Que group（P<0.01）;compared with the blank group,it increased in experimental group and Que group（P<0.05）;compared with the experimental group,the expression of ABCA1protein obviousely increased in Que group（P<0.05）,ABCG1 protein expression was slightly lower,and no statistically significant（P>0.05）;there was significant difference in the four groups by one-way ANOVA（P<0.05）.8.Compared with the control group,the expression of ABCA1 and ABCG1 mRNA decreased significantly in the experimental group and the Que group(P_ABCA1<0.01,P_ABCG1<0.05);compared with the experimental group,it increased in the Que group（P<0.05）;compared with the blank group,the expression of ABCA1 mRNA was slightly increased in the experimental group,there was no significant difference（P>0.05）,however,the expression of ABCG1mRNA was statistically significant（P<0.05）;the difference was statistically significant in the four groups（P<0.05）.9.The differences of IL-6 and TNF-αin the four groups were statistically significant（P<0.001）;compared with the control group,the levels of IL-6 and TNF-αincreased in experimental group and the Que group(P_IL-6<0.05,P_TNF-α<0.01);the levels of IL-6,TNF-αin the Que group were lower than in the experimental group(P_IL-6<0.05,P_TNF-α<0.01);compared with the blank group,the levels of IL-6 and TNF-αin the experimental group and Que group were higher.Conclusion:1.The results suggests the functions of HDL in patients with CHD is damaged,may be one of the mechanism for CHD occurrence.2.According the research results,MPO/PON1 level may indirectly reflect the HDL reverse cholesterol transport function.3.The results shows quercetin can improved reverse cholesterol transport function and inflammatory responses in macrophages,the specific mechanism needs further study.