Mordicin MD28 Increases ABCA1 Expression By Down-regulating MiR-23b-3p

[Objective]To find Momordicin MD28 can upregulate the expression of the preliminary work of THP-1 derived foam cells is reduced ABCA1 lipid accumulation in their cells, but the mechanism is not clear. This study is to analyze the post-transcriptional level and its mechanism.[Methods]First, after using mi R microarray analysis by Momordicin intervention, 50 mg / ml ox LDL for 48 h of mi Rs spectrum THP-1-derived foam cells down 1.5 times mi Rs, and compare with genecards mi Rs ABCA1 regulation of gene expression, to find 2 jointly mi Rs, then predicted target genes ABCA1 relationship between this mi Rs and bioinformatics tools, and luciferase reporter gene system to be verified. q RT-PCR and western blot were used to detect the expression of ABCA1 m RNA and protein levels, q RT-PCR to detect the expression levels of mi R-23-3p, liquid scintillation counting method for the determination of cholesterol efflux, oil red O staining assay intracellular lipid accumulation, the last and then bioinformatics analysis mi R-23b-3p expression and regulation of transcription factors. Statistical analysis was performed using the mean ± standard deviation(± SD), said by GraphpadPrism5.0.1 for data analysis and mapping, select the 95% confidence interval, P <0.05 was considered statistically significant.[Results]mi R microarray analysis found significantly down 69 mi Rs expression displayed on genecards 113 mi Rs in ABCA1 gene as a target, the intersection of two who is hsa-mi R-93, hsa-mi R-642 b, hsa-mi R-23b-3p, hsa- mi R-130 a, hsa-mi R-513a-5p, hsa-mi R-26 a and hsa-mi R-33 a 7 mi Rs, this 7 mi Rs to hsa-mi R-23b-3p momordin MD28 intervention after the biggest drop, reaching 12 times, and the rest are mostly not more than two times, so as to hsa-mi R-23b-3p-depth analysis as Momordicin MD28 down ABCA1 expression of mi R. 3’-UTR of human ABCA1 has three mi R-23-3p binding sites, wherein 255-261 is conserved sites, with ABCA13’UTR binding free energy of-25.3 kcal / mol. ABCA1 gene target validation experiments confirmed that target gene hsa-mi R-23b-3p of. Momordicin MD28 dose(0mg/ml, 0.4 mg/ml, 1.2 mg/ml, 3.6 mg/ml, 5 mg/ml) and time(0h, 6h, 12 h, 24 h, 48h) dependently upregulated ABCA1 m RNA and protein The expression level to 1.2 mg/ml for 12 h most significant. Use hsa-mi R-23b-3p inhibitors attenuated upregulation Momordicin MD28 for ABCA1 and reduce lipid accumulation and increased outflow THP-1 macrophage-derived foam cells intracellular cholesterol. q RT-PCR analysis showed that Momordicin MD28 can increase THP-1 macrophage-derived foam cells hsa-mi R-23b-3p expression level, bioinformatics analysis showed that, MYC, TGFβ, SPⅠ1, Run X2, EGR1, CREB1, ESR2, FXR has binding sites on promoter of hsa-mi R-23b-3p genes.[Conclusions]Momordicin MD28 dose- and time-dependent manner regulate the expression levels of THP-1 macrophage-derived foam cells ABCA1 m RNA and protein, enhanced cholesterol efflux and reduce lipid accumulation, and its all down with and hsa-mi R-23b-3p related.