| PartⅠDetection of expression of the chemerin, adiponectin, and TNFαof adipose tissue in coronary heart disease with non-coronary heart disease patientsObjective Detection of the expression of chemerin,adiponectin,and TNFαmRNA at epicardial adipose in coronary heart disease and non-coronary heart disease patients.Contrast detection of expression of chemerin at chest subcutaneous adipose between coronary heart disease and non-coronary heart disease patients. Detection of expression of chemerin at saphenous vein subcutaneous adipose of coronary heart disease. Analysis the correlation of chemerin expression and coronary heart disease.Methods Collected the tissue samples of epicardial, chest subcutaneous and saphenous vein subcutaneous adipose of cardiac surgery hospital patients with thoracotomy from Dec 2010 to Jun 2011, of which 15 patients with coronary heart disease,13 non-coronary heart disease cases. Detected the expression of the chemerin, adiponectin, and TNFαmRNA at epicardial adipose of coronary heart disease and non-coronary heart disease patients with Real-time quantitative PCR. Contrast detected the expression of cheremin at chest subcutaneous adipose of coronary heart disease and non-coronary heart disease patients. Detected the expression of chemerin at saphenous vein subcutaneous adipose at coronary heart disease patients. Results Compared with non-coronary heart disease, the expression of adiponectin mRNA of coronary heart disease patients’ epicardial adipose decreased, and the TNFa.and the chemerin mRNA Increased. In the same patient, the expression of chemerin at epicardial adipose increased compared with chest subcutaneous adipose, but had no difference with the chest subcutaneous adipose and saphenous vein subcutaneous adipose in coronary heart disease (P<0.05).Conclusion:T the expression of chemerin mRNA of coronary heart disease patients’ epicardial adipose increased。 PartⅡThe establishment of THP-1 macrophage-derived foam and detection of influence of OX-LDL in apoptosis on THP-1 macrophage-derived foam cellsObjective TO establish THP-1 macrophage-derived foam cells and Investigate the appropriate concentration of OX-LDL-induced foam cells.Methods PMA induced THP-1 cell for 48h, observed the appearance of cell morphology and cell phenotype changes, detected the expression of CD68, oil red O staining of intracellular lipid droplet. In different concentrations of oxidized low-density lipoprotein in solution incubated 48 h, detected the total cellular cholesterol content using the enzyme colorimetric assay and the level of apoptosis with flow cytometry.Results In the PMA-induced the TLP-1 cells appeared macrophage-like changes in pseudopods, macrophage marker CD68 expression was positive. Oil red O staining of lipid found in a large number of cells present. Oxidized low-density lipoprotein concentration(25～50ug/mL), no significant apoptosis. With oxidized low-density lipoprotein concentration increased, the rate of apoptosis and necrosis rate also increased. Total cholesterol levels of Cells was detected by ELISA and found that oxidized low-density lipoprotein concentration (25ug/mL to 50ug/mL), the total intracellular cholesterol content increased, the concentration of 100ug/mL then began to decline.Conclusion THP-1 macrophage-derived foam cells were macrophage-like changes in pseudopods, expressed CD68-positive. Cells had a large number of lipid, oil red O staining positive. Oxidized low-density lipoprotein concentration 50ug/mL macrophages constructed expression levels of total cholesterol is better, apoptosis is lower, the optimal concentration for the induction. PartⅢThe influence of expression of CD36, ABCA1 inflammatory cytokines TNFα, and the cholersterol accumulation treated with chemerin at THP-1 macrophage-derived foam cells.Objective The influence of CD36, ATP-binding cassette transporter Al,and inflammatory cytokines TNFαand change at intracellular cholersterol accumulation of THP-1 macrophage-derived foam cells at different concentrations of adipokine chemerin. Explore the possible mechanism of chemerin regulation of intracellular lipid.Method Combined different concentrations of Chemerin with the same concentration of oxidized low-density lipoprotein -induced THP-1 macrophage-derived foam cells (50ug/mL)24h, detected intracellular levels of total cholesterol, and CD36, ABCA1 mRNA and protein expression with Western Blot and RT-PCR. Detected the expression. of TNFαin the culture medium by ELISAResult Different concentrations of Chemerin-stimulated intracellular expression of total cholesterol levels were increased, the more obvious concentration decrease, and showed dose-dependent. The expression CD36 and expression of ABCA1 protein decreased, the expression of TNFαin medium increased.Conclusion Chemerin possibly increased expression of CD36, reduced ABCA1 mRNA and protein expression, increased THP-1 macrophage-derived foam cells cholesterol accumulation, and reduced expression of CD36 and ABCA1 may be associated with increasing of the release of pro-inflammatory cytokines TNFαstimulated by Chemerin of THP-1 macrophages source foam cells. PartⅣChemerin stimulated THP-1 macrophage-derived foam cells on the NF-κB, LXRα,, PPARγsignaling molecule expressionObjective:Detection expression levels of the cells NF-κB, LXRα, PPARγsignaling molecule protein in THP-1 macrophage-derived foam cells in different concentrations of Chemerin intervention. Invested the possible mechanisms of Chemerin inhibited THP-1 macrophage-derived foam cells in ABCA1 of expression.Method:Incubated with different concentrations of Chemerin in THP-1 macrophages-derived foam cells for 24h,, the content of LXRα, PPARγprotein content changes, detection the content of nuclear and cytoplasmic NF-κB (P65) of with Western Blot.;Results:Chemerin stimulated THP-1 macrophage-derived foam cells after 24h, increased foam cell NF-κB activity, PPARγexpression and, LXRαprotein expression was not significantly change。Conclusion:Chemerin inhibts ABCA1mRNA expressing may be caused by enhancing of NF-κB activity... |
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