Isolation of Prismatomeris Connata Y.Z.Ruan compounds and their effects on LPS-induced inflammation in RAW264.7 cells
Objective To study the effect of Prismatomeris Connata Y.Z.Ruan extract on the lipopolysaccharide(LPS)-in-duced RAW264.7 cell inflammation model,and to isolate and purify the active ethyl acetate fraction.Methods CCK-8 method was used to detect the effects of chemical components of Prismatomeris Connata Y.Z.Ruan root on RAW264.7 cell activity.RAW264.7 cells induced with LPS at a concentration of 500 ng/mL were used to establish a cell inflammation model.Griess as-say was used to detect the level of NO in the cell supernatant;ELISA was used to detect the content of IL-1β and TNF-α in the cell supernatant;DCFH-DA fluorescent probe was used to detect the level of ROS in the cells.The ethyl acetate fraction of Prismatomeris Connata Y.Z.Ruan root was isolated and purified by using solvent extraction,silica gel column chromatography,and high-performance liquid chromatography(HPLC)methods.Results Prismatomeris Connata Y.Z.Ruan extract,petroleum ether fraction,ethyl acetate fraction,n-butanol fraction,and water fraction at a mass concentration of 100 μg/mL and below showed no toxicity to RAW264.7 cells.Compared with the model group,Prismatomeris Connata Y.Z.Ruan extract and ethyl acetate fraction had a down-regulating inhibitory effect on the expression levels of NO,TNF-α,IL-1β and ROS in the cells.Based on the physicochemical properties and spectroscopic data,four compounds were isolated and identified from the ethyl ace-tate fraction of Prismatomeris Connata Y.Z.Ruan extract which were methyl isalizarin-1-methyl ether(1),1,3-dihydroxy-2-ethoxymethylanthraquinone(2),di(2-ethylhexyl)phthalate(3),di(2-ethylhexyl)terephthalate(4).We did not discuss the isolated monomer which did not have anti-inflammatory activity.Conclusion The ethyl acetate fraction and the Prismatomeris Connata Y.Z.Ruan extract can antagonize the LPS-induced inflammatory response in mouse macrophages and inhibit the expres-sion of inflammatory factors.Di(2-ethylhexyl)phthalate and di(2-ethylhexyl)terephthalate were isolated from this plant for the first time.
root of Prismatomeris Connata Y.Z.RuanlipopolysaccharideRAW264.7 cellinflammationNOROSin-flammatory cytokine
国家科技期刊平台
NSTL
万方数据
