Study On The Chemical Constituents And The Anti-inflammatory And Immunological Activities Of Urtica Fissa

Urtica fissa E.Pritz.a perennial herb,has a long history to be used as traditional Chinese medicine,the Minority has used its water decoction for the treatment of rheumatoid arthritis,diabetes and skin eczema.However,there are few reports about the research on active ingredients against rheumatoid arthritis and the research on chemical constituents of U.fissa.Meanwhile,U.fissa also lack of quality control indicators,thus clarifying the material base of U.fissa against rheumatoid,has a very important significance.Prefered studies of our laboratory have shown that Urtica fissa has anti-rheumatoid component.The aim of this experiment is to study the chemical constituents of U.fissa further based on the previous studies.To verify the anti-rheumatoid arthritis and the immunological activity of U.fissa by the in vitro activity experiment.Thus,to determine the anti-rheumatic and the immunological active ingredients of U.fissa,to clarify the basis of anti-rheumatoid material of U.fissa and laid the foundation.The chemical constituents of U.fissa were purified by the methods of macroporous resin column chromatography,silica gel,reverse phase silica gel column chromatography,Sephadex LH-20 column chromatography,preparative high performance liquid chromatography and recrystallization.A total of 38 compounds were isolated and identified from the ethanol extract of U.fissa by the modern spectroscopy combined with physicochemical properties.They were Urticin A(1),Urticin B(2),Urticin C(3),Urticin D(4),Urticin E(5),Urticin F(6),Urticin G(7);cordyrrole A(8),3-Hexenedioic acid,monomethyl ester(9),stearic acid(10),mannitol(11),vanillic acid(12),5-hydroxyl-2-hydroxymethyl pyridine(13),macrophyllilactone E(14),uracil(15),quercetin(16),cleomiscosin A(17),caffeic acid(18),chlorogenic acid(19),p-hydroxybenzaldehyde(20),ficusol(21),crinumaquine(22),2-[formyl-5-(methoxymethyl)-1-hydro-pyrrol-1-yl] butyric acid butyrate(23),2-[formyl-5-(butyl ether methyl)-1-hydrogen-pyrrol-1-yl] butyric acid 15 butyl ester(24),4-[formy-5-(methoxymethyl)-1H-pyrrol-1-yl]butanoic acid(25),2-[formyl-5-(methylbutyl ether)-1-hydrogen-pyrrol-1-yl] butyric acid methyl ester(26),2-[formyl-5-(methoxymethyl)-1-hydro-pyrrol-1-yl] butyric acid butyl ester(27),lobechine(28),erythritol(29),methyl ?-D-galactopyranoside(30),(6R,9R)-Roseoside(31),urticol-7-O-?-D-glucopyranoside(32),bis(4-hydroxyphenyl)methanol(33),pinoresinol monomethyl ether-4-O-?-D-glucoside(34),kaempferitrin(35),adenosine(36),capparisine A(37),capparisine B(38).Among them compounds 1-7 were new compounds,compounds8?9?11-14?17?20-35?37?38 were first isolated from Urtica.The MTT assay was used to detect the effects of different polarities and compounds of ethanol extract of U.fissa on RAW264.7 cell viability.The effects of different polarities of ethanol extract of U.fissa on LPS-induced NO content in culture supernatant of RAW264.7 cells were determined by Griess method.Results were as follows(1)Every extraction and compounds from ethanol extracts of U.fissa all showed few cytotoxicity on RAW264.7 cells;(2)The inhibitory effect of NO was obvious on RAW264.7 cell culture supernatant induced by LPS.In the ethanol extract of U.fissa,with the increase of the extraction polarity,the cytotoxicity was decreased and the inhibition of NO was decreased.These results maybe suggest that U.fissa has some anti-inflammatory activity to resist rheumatoid arthritis.Alkaloid compounds,glycosides,lignans compounds show a more significant anti-inflammatory activity.The results show that U.fissa has a certain anti-inflammatory activity,to achieve the role of anti-rheumatoid arthritis.The anti-complement activity of the ethanol extract of U.fissa at different polarities was determined by the classical method,the results were the non-polar parts have more stronger anti-complement activity,while the polar parts were not so strong.Dispite the water extract,the other extracts all has anti-complement activity.It also well explained that U.fissa may achieve anti-rheumatoid arthritis activity by anti-complement activity.Our experiment have studied the chemical composition of U.fissa for the first time,and studied the anti-inflammatory and anti-complement activity systematically.A total of 38 compounds were isolated and identified from the ethanol extract of U.fissa.Among them 7 compounds were new compounds,and has anti-inflammatory active,and 25 compoundswere first isolated from U.fissa.Clarify the active ingredient of U.fissa against rheumatoid arthritis and provided scientific basis for the further study of U.fissa.