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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 CD4+ 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 24 hours and co-cultured with mouse CD4+ T cells for 72 hours. For the determination of T-bet, GATA-3, RORγt, Foxp3, DLL-3, and Jagged-1 mRNA gene expression, the floating cells (CD4+ 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-3 gene but suppressed Jagged-1 gene expression in oxLDL-treated BMDC. Furthermore, GP ethanol extract and its fractions suppressed T-bet, GATA-3, and RORγt gene expression but increased the expression of the Foxp3 gene in differentiated CD4 + 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 Th1, Th2, and Th17 cells while increasing anti-atherogenic cells, Treg cells.

Introduction: Gynura procumbens (GP) is a medicinal plant with numerous beneficial pharmacological activities. The aim of this study was to identify the bioactive phytoconstituents in GP ethanol extract and hexane, chloroform, ethyl acetate, and aqueous fractions of GP, and also to evaluate the cell viability of GP ethanol extract and its fractions-treated RAW264.7 cells.Methods: The ethanol GP leaf extract was prepared and further subjected to fractionation. The cell viability of GP ethanol extract and its fractions-treated RAW264.7 cells were measured by PrestoBlue. The phytoconstituents of GP ethanol extract and its fraction were determined by using liquid chromatography-mass spectrometry (LC-MS).Results: RAW264.7 cells exposed to the GP ethanol extract and its fractions showed significantly high proliferation and weak cytotoxic effect on the macrophages, with an average inhibitory concentration of 90% at 24, 48, and 72 hours of incubation. However, at a concentration of 10 μg/mL, the aqueous GP fraction clearly displayed anti-proliferative properties because the cell viability of aqueous GP fraction-treated RAW264.7 cells reduced to 64%, 29% and 4% after 24, 48 and 72 hours of incubation, respectively. The GP extracts and its fractions contained mainly fatty acids, flavonoids, sesquiterpenoids, and products of chlorophyll breakdown.Conclusion: GP ethanol extract and its fractions at certain concentrations may act as immunomodulators, as they induced promising proliferation activity of macrophages. Further studies are needed to determine either the identified chemical compounds influenced on the proliferation of macrophages solely or cooperatively.