Research About Antiinflammatory Constituents And Fat-soluble Constituents From Korean Medicine The Seeds Of Lepidium Apetalum Willd

North Semen Lepidii is dry mature seed of Lepidium apetalum Willd.,which is belong to Brassicaceae family.It has antibacterial,anticancer,lipid regulating and other pharmacological effects.In order to further research and develop the application of the drug,the chemical constituents and anti-inflammatory activities of the seeds of Lepidium apetalum Willd.were mainly studied in this experiment,in order to give full play to its medicinal value and provide scientific basis.In this experiment,70%ethanol was selected to heat and circumfluence the seeds of Lepidium apetalum Willd.concentrated and dried to obtain the extract,then degreased with petroleum ether,and then extracted with ethyl acetate to obtain the ethyl acetate layer.The ethyl acetate layer was separated and purified by Sephadex LH-20 chromatography,silica gel column chromatography,and preparative medium pressure liquid chromatography.Ten compounds have been identified by using a variety of spectroscopy methods and references.Compounds respectivelyβ-Sitosterol(1),Daucosterol(2),Isorhamnetin(3),Isorhamnetin-3-O-β-D-glucopyranoside(4),Quercetin-3-O-β-D-glucopyranoside(5),Sinapic acid(6),1(E),2(E)-O-Di-sinapoyl-β-D-glucopyranoside(7),I-Monopalmitin(8),1-Monstearin(9),Kaempferol-3-O-β-D-glucopyranoside(10).The anti-inflammatory activity of the compound was determined in vitro using lps-induced mouse macrophage RAW264.7 as an inflammatory model.Compounds 7 and 8 significantly reduced the content of NO released by lps-induced RAW264.7 cells at concentrations of 20 μM and 40 μM,showing strong anti-inflammatory activity.Compounds 3,5,8 and 9 were isolated from the seeds of Lepidium apetalum Willd.for the first time.Compounds 8 and 9 is the first report from the genus Lepidium.Compound 9 is the first report from the family Brassicaceae.At present,the Pharmacopoeia does not include data on the content standards of the seeds of Lepidium apetalum Willd.And compound 4 was found to be the main component in the process of extraction and separation of the seeds of Lepidium apetalum Willd.Therefore,the HPLC method was used to determine the content of Isorhamnetin-3-O-β-D-glucopyranoside,which provided reference for the content standard of the seeds of Lepidium apetalum Willd.in Pharmacopoeia.Isorhamnetin-3-O-β-D-gIucoside has a good linear relationship at the level of 10.0-500.0 μg/mL,Y=8088.8X+35308(R2=0.9991),the RSD values of precision,stability,repetitiveness test were all less than 2%(n=6),sample adding recovery is 100%(RSD=1.23%).Thus,HPLC method could accurately determine the content of Isorhamnetin-3-O-β-D-glucoside in the seeds of Lepidium apetalum Willd.The results showed that the content of Isorhamnetin-3-O-β-D-glucoside in the seeds of Lepidium apetalum Willd.was 7.5mg/g.There are south and north parts of Semen Lepidii,due to the different habitats,the seed of Descurainia sophia(L.)Webb.ex Prantl.is South Semen Lepidii,which is belong to Brassicaceae family,and mainly distributed in Jiangsu,Anhui,Zhejiang and other places.The seed of Lepidium apetalum Willd.is North Semen Lepidii,which is belong to Brassicaceae family,and mainly distributed in Jilin,Liaoning,Inner Mongolia,Hebei and other places.Because of their different pharmacological effects,may be related to the different composition and content of the ingredients.In order to analyze the differences of composition and content of liposoluble components in Semen Lepidii from different habitats,the pharmacological effects of the two are different,which may be related to the composition and content of components.In order to analyze the differences in composition and content of fat-soluble of Semen Lepidii from different habitats,the fat-soluble of south Semen Lepidii from Anhui,north Semen Lepidii from Hebei and north Semen Lepidii from Yanbian were tested by GC-MS.Results show that different habitats of Semen Lepidii fat-soluble composition and content all have differences,seventy compounds were identified in the fat-soluble components of Semen Lepidii from three different habitats.34,32 and 43 kinds of the fat-soluble components were identified in Semen Lepidii from Anhui,Hebei and Yanbian respectively.The contents of these compounds were all slightly different in the three habitats.The most content was methyl linolenate.The methyl linolenate in Semen Lepidii from Yanbian was 48.67%and had the highest relative content among three habitats.Therefore,it is speculated that Semen Lepidii from Yanbian,as a kind of Korean medicine,may have better effect of lowering blood lipid.