Studies On Chemical Constituents And Pharmacological Effect Of Active Fraction For Fracture Healing In Sambucus Williamsii Hance

Sambbucus williamsii Hance belongs to the genus Sambbucus chinenis. As a species of the genus, Sambbucus williamsii Hance is also called Xu-gu-mu, Da-jie-gu-dan, Gong-lao-dao, etc. Sambbucus williamsii Hance is a folk medicine with a long history of safe use for treatment of fracture and joint disease in China. However, the active constituent of Sambbucus williamsii Hance and the effects of Sambbucus williamsii Hance on bone fracture remain largely unknown. In this study, the components obtained from the stems of Sambbucus williamsii were elucidated. Besides, the effects of the components on bone fracture and their mechanisms of action were examined in vivo and in vitro.At first, we identified the bioactive constituents from the 50% ethanol extract of the stems of Sambbucus williamsii. Then, the effects of isolated compounds on bone fracture, hemorrheology, blood biochemistry, anti-inflammation and analgesia were evaluated. The results showed that the active components markedly improving bone fracture recovery. Treatment with isolated compounds exerted a significant elevation in serum level of AIP and BGP and they showed marked anti-infflamatory effect, analgesic effect as well. Administration of the active components markedly ameliorated hemorheology when compared with model.In this study, isolation procedure was carried out by column chromatography and HPLC;structures of isolated compounds were identified by ¹H-NMR, ¹³C-NMR, ¹H-¹HCOSY, HSQC, HMBC and ESI-MS. Isolation of the 50% ethanol extract of the stems of Sambbucus williamsi afforded 23 constituents, including 12 iridoids compounds, 10 lignans compounds, 1 phenylpropanol compound, in which 5 iridoids compounds and 4 lignans compounds were novel compounds and 12 compounds were firstly found in Sambucus williamsii Hance . The structures of the nine novel compounds were identified as Williamsoside A(7- β -O-Ethylepimo-rroniside),Williamsoside B (7β -O-ethylmorroniside-(6'-O-7")- β -morroniside), Williamsoside C ( α -D-glucopyranosyl(1→2)-D-fructofuranosyl(4→6)- β -morroniside ),Williamsoside D ( α -D-apiofuranosyl (1"→6' )-7-deoxyloganin),Williamsoside E ( α-D-apiofuranosyl(1"→6')-loganin) ,Williamsoside F ((7R,8R)-4,9-dihydroxy-3-methoxy-4'.- 7, 5': 8-benzodioxanelignan-9'-O-ï¿¡-D-glucopyrano-side) ,Williamsoside G ((7S,8R)-4,9-dihydroxy-3,3'-dimethoxy-7,8dihydrobenzo-furan-1' -propylneolignan-9' -O-[ a -D-apiofuranosyl (1—6)]- ï¿¡ -D- glucopyrano-side) ,Williamsoside H (4,7,9,3' -tetrahydroxy-3- methoxy-8-0-4' isolignan-9' -O-0 -D- glucopyranoside (thero)) and Williamsoside I.Then in vitro experiments were performed to determine the effects of morroniside on proliferation, differentiation and mineralization of MC3T3-E1. Results suggested morroniside markedly improved the proliferation, differentiation and mineralization of MC3T3-E1. At the same time, the effects of morroniside on the level of Ca2+ in cell were investigated by confocal laser scanning microscope. Results showed that morroniside significantly increased the level of Ca2+ in cell. In conclusion, this study identified the bioactive components of Sambbucus williamsii Hance and investigated the effects of the bioactive components on bone fracture and the mechanisms involved in it. It is undoubted that this study not only affords an evidence to discover a new folk medicine to treat bone fracture but also improve the cognition for bone fracture recovery from pharmacological point.