Antiosteoporotic Activity And Metabolomics Of Cistanche Deserticola Active Constituents

Objective: To establish HPLC fingerprint and content determination methods for qualitative and quantitative analysis of Cistanche deserticola Y.C.Main Ningxia;ovariectomized rat(mice)modelswere used to evaluate the therapeutic effects of Cistanche deserticola extract,total phenylethanoid glycosides fraction and phenylethanoid glycosides components;the metabolomics technique was applied to estimatethe possible mechanism of the anti-osteoporosis,by using UPLC-QTOF analysis.Methods: 1.With echinacoside as a reference peak,and acteoside and other four ingredients as index components,to establish the fingerprint of HPLC-DAD.2.The pathological state of postmenopausal osteoporosis was simulated with ovariectomizedrats(mice).The ovariectomized animals were orally administered with Cistanche deserticola extract(800,400,200 mg/kg),total phenylethanoid glycosides fraction of Cistanche deserticola(240,120,60 mg/kg),Cistanche A and acteoside(80,40,20 mg/kg),2?-acetylacteosideand 6?-acetylacteoside(40,20,10 mg/kg),respectively;after 12 weeks of continues orally administrated intervention,the osteoporotic-related bone mineral density and bone microarchitecture,as well as bone formation and bone resorption index were evaluated.3.The serum of the above ovariectomized animals were used for UPLC-QTOF analysis,and PCA and PLS-DA statistical methods were employed to discovery the differential metabolites based on p<0.05 and VIP>1.Metabolites structure was determined by using the literature,database and standard control.Results: 1.The HPLC fingerprint for qualitative analysis of Cistanche deserticola was established.2.Cistanche deserticola extract,total phenylethanoid glycosides fraction and phenylethanoid glycosides components can improve the bone mineral density,bone microarchitecture,increase bone formation indext and reduce bone resorption index in ovariectomized rats(mice).3.33 endogenous metabolites including valine were screened by the database and literature from positive and negative ion modes.When compared with sham operation group,leukotriene F4,PC(22:6/18:1),PC(40:6),PI(32:0),PC(32:1),lysophosphatidylcholine(20:3),LPE,LPC18:0a,LysPc20:4,sphingosine 1-phosphate,taurocholic acid,SM,Lphenylalanine,Dioleoylphosphatidylcholine,valine,N-methyl-amino isobutyric acid and indole-3-acetamide,25-hydroxy vitamin D3,linolenic acid,phytosphingosine in OVX rats were significantly reduced,while carnitine,2-methyl malonic acid,5-hydroxy-N-benzoyl methyl guanine,serine,quinine acid and sugar with deoxycholate 3-glucuronide deoxyadenosine,3-hydroxy benzene acetic acid and 5-HIAA were significantly elevated,the total phenylethanoid glycosides of Cistanche deserticolacan improve the level of leukotriene F4,PC(22:6/18:1),PC(40:6),PI(32:0),PC(32:1),lysophosphatidylcholine(20:3),LPE,sphingosine 1-phosphate,taurocholic acid,SM,two L-phenylalanine,oleoyl phosphatidylcholine,phenyl pyruvic acid,valine,carnitine,2-methyl malonic acid,5-hydroxy-N-benzoyl methyl guanine,methylhippuric acid,quinic acid and Glycochenodeoxycholicacid 3-glucuronide,deoxyadenosine,3-Hydroxyl phenylacetic acid and 5-HIAA.The mechanisms of action were mainly through the intervention of fatty acid,amino acid and phospholipid metabolism.Conclusion: 1.The HPLC fingerprint can be used for the qualitative analysis of Cistanche deserticola.2.Cistanche deserticola extract,total phenylethanoid glycosides effective parts and active ingredients have the therapeutic effects on osteoporosis induced by ovariectomy in rats(mice).3.Cistanche deserticola may play an important role in the treatment of osteoporosis by adjusting the 33 metabolites such as valine.The mechanisms were mainly related to fatty acid,amino acid,and phospholipid metabolism.