Gynura procumbens ethanol extract and its fractions inhibit Th۱, Th۲ and Th۱۷ but induce Treg cells differentiation during atherosclerosis development

Introduction: Gynura procumbens (Lorr.) Merr. (GP) displays cardio-protective effect, which may hinder atherogenesis induced by oxidized low-density lipoprotein (oxLDL) and leukocytes. The current study was undertaken to elucidate the chemical constituents of GP ethanol extract and its aqueous, chloroform, ethyl acetate, and hexane fractions, and their effects on CD۴+ T cell differentiation during atherogenesis. Methods: Initially, the bioactive constituents in GP ethanol extract and its fractions were analyzed using gas chromatography-mass spectrometry (GC-MS). Generated mouse bone marrow dendritic cells (BMDC) were loaded with oxLDL and GP ethanol extract and its fractions for ۲۴ hours and co-cultured with mouse CD۴+ T cells for ۷۲ hours. For the determination of T-bet, GATA-۳, RORγt, Foxp۳, DLL-۳, and Jagged-۱ mRNA gene expression, the floating cells (CD۴+ T cells) and adherence cells (BMDC) were isolated from their total RNAs and reverse transcribed into cDNA.Results: GC-MS analysis showed that GP ethanol extract and its fractions contained various volatile compounds. GP ethanol extract and its fractions also increased the DLL-۳ gene but suppressed Jagged-۱ gene expression in oxLDL-treated BMDC. Furthermore, GP ethanol extract and its fractions suppressed T-bet, GATA-۳, and RORγt gene expression but increased the expression of the Foxp۳ gene in differentiated CD۴ + T cells. Conclusion: GP ethanol extract and its fractions are composed of various bioactive chemical components that can induce anti-atherogenic effects by inhibiting pro-atherogenic cells such as Th۱, Th۲, and Th۱۷ cells while increasing anti-atherogenic cells, Treg cells.