Anti-osteoporotic Chemical Constituents In Curculigo Orchioides And It's Quality Evaluation

Curculigo orchioides is the dry rhizome of Curculigo orchioides Gaertn,one of the traditional Chinese medicine and had been included in the last two editions of Chinese pharmacopeia.In traditional Chinese medicine,Curculigo orchioides are considered to have the effects of maintaining healthy energy,and nourishing the liver and kidneys.People usually use Curculigo orchioides to treat bone metabolism disorder disease and osteoporosis in some Chinese herbal formulae.The main constituents of Curculigo orchiodes are cycloartane-type triterpene saponin,phenol and phenolic glycoside,lignan and lignan glycoside,flavone,etc.The main potential pharmacological activities of Curculigo orchioides had been reported are immunostimulatory,antioxidant,taste-modifying,liver and cardiovascular system protection, tonifying kidney and antiosteoporosis,etc.Although Curculigo orchioides had been widespreadly used in the presently clinical antiosteoporotic formula,the base of its active compounds is not clear??our study,after we confirmed the antiosteoporotic activity of ethanol extract of Curculigo orchioides by OVX model,we isolated the chemical constituents from it and looked for its antiosteoporotic compounds by osteoblast and osteoclast cell culture in vitro. Meanwhile,we carried out quality evaluation on Curculigo orchioides collected from different places by HPLC-UV analysis.We used OVX model to investigate the antiosteoporotic activity of the ethanol extract of Curculigo orchioides,after experiment we concluded that administration of Curculigo orchioides extract prevented bone loss in the trabecular bone of the tibia in ovariectomized rats without affecting the weight of the body and the uterus,and increased serum phosphorus,calcium,and OPG levels,decreased serum DPD/Cr, TRAP,ACTH,and corticosterone levels,but did not alter serum TNF-α,IL-6,and ALP levels in ovariectomized rats.Curculigo orchioides ethanol extract has a definite protective effect on bone loss in ovariectomized rats by inhibiting bone resorption and increasing serum phosphorus and calcium levels, without affecting bone formation.Therefore,Curculigo orchioides can be considered a potential antiosteoporosis herbal plant,although more studies are needed to clarify its real potential chemical constituents and their mechanism of action.Seven compounds were isolated from Curculigo orchioides by the conventional chromatography and semi-preparative HPLC and all seven compounds were identified by UV,IR,MS,¹H-NMR,¹³C-NMR, 2D-NMR.Among them,a novel chlo-contain phenol and six known compounds were islosted.They are 2,6-dimethoxy benzolic acid(1),curculigoside A(2) and B(3),curculigine A(4),4- dichlorine-5-hydroxyl-3-methylphenol-l-O-β-D-glucopyranosyl-(1→6)-β-D-glucopyranosede,and named as curculigine D(5),3,3',5,5'-tetramethoxy-7,9':7',9-diepoxylignan -4,4'-di-O-β- D-glucopyranoside(6),andβ-sitosterol respectively.Compound 5 is a new chloro-contained type phenol isolated from Curculigo orchioides with the resemble structure as curculigine A.After the screening by primary osteoblast and induced osteoclast,the phenolic glycosides have potential antiosteoporotic activity,they can potentially stimulate osteoblast proliferation and inhibit osteoclast TRAP activity.12 samples were collected from different places of southwest district in China,we carried out the fingerprint research about them and established a new quality evaluation way for estimating their quality by determining the content of curculigoside A and B.The results revealed that the chemical constituents of these samples were obviousely different,and the curculigoside A and B content also differed in these sample?? the samples the content of curculigoside A was highest in CO-ES(0.538%) and lowest in CO-WS(0.028%),the content of curculigoside B was highest in CO-YL(0.330%) and lowest in CO-BN(0.103%).The samples were collected from the range of geographic places,east longitude 102.06 to 106.87,northern latitude 23.15 to 30.11,altitude 315m to 1850m,the chemical constituents of different samples may related to the factor of geography,climate,collect time,age,etc.We also determined the curculigoside A and B in 6 good samples bought from drugstores of 6 different provinces in China,the differences of curculigoside A and B content were also significant,we can presume that the chemical constituents content may related to their processing thecnology and store way.Curculigoside A and B are the main constituents in the collection and goods samples,it can be more accurate to estimate its quality by determining these two compounds simultaneously.A way that estimate the quality by determining more main chemical constituents is very important to estimate medicine quality,we need to do more efforts to establish a more accurate way to control the quality of Curculigo orchioides and find out its best grow place.