Keywords

Momordica Charantia Protein Adenosine Triphosphate-Binding Cassette Transporter A1 Hsa-Mir-23B-3P Lipid Accumulation

Abstract

Objective Preliminary work found that bitter melon protein (MD28) can up-regulate the expression of adenosine triphosphate-binding cassette transporter A1 (ABCA1) in THP-1-derived foam cells and reduce intracellular lipid accumulation, but its mechanism is unclear. This article intends to analyze its mechanism of action from the post-transcriptional level. Methods: First, the miRNA chip technology was used to analyze the 1.5-fold down-regulated microRNA (miRNA) in THP-1-derived foam cells treated with 50 mg/L ox-LDL for 48 h after MD28 intervention, and combined with Genecards to regulate ABCA1 gene expression. Compare the miRNAs to find the common ones, and then use the luciferase reporter gene system to verify their target gene relationship. qRT-PCR was used to detect the expression levels of ABCA1 mRNA and miR-23b-3p, Western blot was used to detect protein expression levels, and Oil Red O staining was used to determine intracellular lipid accumulation. Results: miR-23b-3p is the intersection of miRNA chip and Genecards, and the target gene verification experiment confirmed that ABCA1 is the target gene of miR-23b-3p. MD28 dose (0 g/L, 0.4 g/L, 1.2 g/L, 3.6 g/L, 5 g/L) and time (0 h, 6 h, 12 h, 24 h, 48 h)-dependent upregulation The expression levels of ABCA1 mRNA and protein were most significant when treated with 1.2 g/L for 12 h. Ox-LDL upregulates the expression of miR-23b-3p and is inhibited by MD28. MD28 can reduce intracellular lipid accumulation. Inhibitors of miR-23b-3p can antagonize the effect of MD28 on ABCA1 expression and intracellular lipid accumulation. Conclusion MD28 mediates its effect on promoting ABCA1 expression and reducing intracellular lipid accumulation in THP-1-derived foam cells by downregulating miR-23b-3p.

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Original research done by Yang Lu, Wang Zuo, Wang Zhuo, Wang Yan, Fei Mingzhu, Chen Jiaojiao, Ma Xiaofeng

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