Effects Of Pioglitazone On The Gene Expression Of ABCA1,ABCG1inTHP-1Derived Foam Cell And Cholesterol Outflow

Objective To explore Pioglitazone effects on gene of ATP binding cassette transporter Al, ATP binding cassette transporter G1expression in THP-1derived Foam cell and Cholesterol levels, so as to explore theory of action of pioglitazone resistance to hardening of the arteries.Methods1. Establish THP-1derived foam cell model. THP-1cells were cultured and induced to differentiate into macrophages by treatment OX-LDL macrophages to produce foam cells, oil red O staining of foam cell formation. According adding different concentrations of pioglitazone into10,20,30umol/L pioglitazone intervention group and the control group (saline added). Using high performance liquid chromatography analysis of intracellular total cholesterol (total cholesterol, TC), free cholesterol (free cholesterol, FC) and cholesterol ester (cholesterol ester, CE) content.(3) with real-time quantitative PCR method was measured24hours ABCA1, expression of mRNA of ABCGI; application Western-blot assay24hours ABCA1, ABCG1protein expression levels.4using SPSS17.0statistical software for statistical analysis. Experimental data are mean±standard deviation () that differences between groups were tested using ANOVA and SNK, P <0.05was considered statistically significant.Results1. THP-1cells in vitro after PMA induced to differentiate into macrophages; macrophages then after ox-LDL-induced oil red O staining bright red lipid droplets in cells, which means that successful differentiation of macrophages into foam cells,the control group staining was significantly highter than ox-LDL+pioglitazone group (Figure1)2. Real-time PCR to detect different concentrations (10umol/l,20umol/l,30umol/l) mRNA expression24hours of pioglitazone group THP-1derived foam cells ABCA1were2.04±0.047、3.21±0.168、5.34±0.094There was a significant difference in comparison (P<0.05) and the control group (1.00±0.004). MRNA expression24hours with different concentrations of pioglitazone group THP-1derived foam cells ABCG1were2.08±0.011、3.34±0.52、5.60±0.103and the control group (1.00±0.002) were significantly different compared (P<0.05)(table3)3. Westernblot different concentrations (10umol/l,20umol/l,30umol/l) Expression pioglitazone group THP-1derived foam cells in24hours ABCA1protein were1.97±0.02,5.58±0.04,3.98±0.04There were significant differences in comparison (P<0.05) and the control group (1.00±1.97±0.02、3.98±0.04、5.58±0.040.003). The expression of different concentrations of pioglitazone group THP-1derived foam cells24hours ABCG1protein were2.06±0.04、3.62±0.07、5.92±0.07and the control group (1.00±0.004) were significantly different compared (P<0.05)(table4).4. Using high performance liquid chromatography analysis of intracellular cholesterol esters, free cholesterol and total cholesterol Show:Compared with the control group, different concentrations of pioglitazone group FC and TC were significantly lower (P<0.05), and showed a concentration-dependent manner.Conclusions1.pioglitazone into ABCA1, ABCG1expression at the mRNA and protein levels, and has a concentration-dependent manner.2.pioglitazone can reduce intracellular cholesterol levels, promote the role of cholesterol efflux.