Studies On The Activity Of Inhibiting Osteoclasts And The Chemical Constituents About Fuctus Toosendan

Fuctus Toosendan are dry fruits of Melia toosendan Sib.et Zucc.in Meliaceae,and mainly grow in Sichuan,Guizhou,Hubei,Hunan,Henan province and the southern part of Gansu province.Fuctus Toosendan have discharging liver,promoting qi circulation to relieve pain and deworming.It is traditionally used for chest and hypochondrium,abdominal distention,colic and enterozoic abdominalgia,et al.Extracts were obtained with the ethanol of 70%from Fuctus Toosendan,and the polarity was separated by the means of systematic solvent extraction.The chemical constituents of the chloroform layer were studied and 7 compounds were isolated with silica gel columns,preparative TLCs,recrystallization etc.On the basis of 1H-NMR, 13C-NMR,MS data,physicochemical characters and reported in literature,6 chemical structures of them were elucidated as:15-triacontanol（Ⅰ）,stigmasterol（Ⅱ）,β-sitosterol（Ⅲ）,△^5,6-Iso-Toosendanin（Ⅳ）,Pentadecane pelargonate（Ⅴ） and 2-（13-methyl）-tridecane caproate（Ⅵ）,and new chemical compound isⅣ,Ⅰ,Ⅱ,Ⅴ&Ⅵare isolated from Fuctus Toosendan.The experimental method of culturing osteoclasts in vitro was established. RANKL could induce RAW cells into osteoclast cells,so monocytes RAW 264.7 cells were used to generate osteoclasts in tissue culture system in the presence of RANKL. The oesteoclasts cells were detected as indicated by TRAP staining.The bioactivity of Fuctus Toosendan inhibiting osteoclasts was studied,and the effective part was determined by means of chemical separation combined with activity detecting.The results display the extract of the ethanol layer has remarkable activity of inhibiting osteoclasts.Meanwhile,the effective part was further purified by macroporous resin,and obtained two active priciples which are the ethanol parts of 40%～60%& 60%～80%.Moreover,the part of 40%～60%has good bioactivity.The inhibition ratio of osteoclast is exceed 95%,even if high- concentration RANKL induces.The finding about inhibiting osteoclasts is the first report,and provid theoretical and experimental bases for the more research and development of Fuctus Toosendan in the future.