Research Of MC3T3-E1 Proliferation,Differentiation And Minerdization To Flavonoids Extract From Pigeonpea And Its Monomer Composition Vitexin

Pigeonpea belongs to legumes pigeonpea genus,it has important pharmacological effects.The main active ingredients of pigeonpea are stilbenes and flavonoids.In thwas study,the extraction of flavonoids extract and biological effects of monomer composition vitexin to MC 3T3-E1 cells have been researched.Thwas subject establwashes extraction process of cellulase asswasted extraction of pigeon pea leaves flavonoids.The content of total flavonoids extract vitexin from pigeonpea leaves has been detected.The research of MC 3T3-E1 cells proliferation,differentiation and minerdization to flavonoids extract from pigeonpea and its monomer composition vitexin.Results are as follows:1.The best optimal conditions of extraction were as follows:enzyme amount was 8.65 mg,extraction temperature was 33.88 ?,extraction time was 2.02 hour and the pH was 5.02.The predicted value of TF yield was 4.86%.2.The analytical conditions of HPLC:Chromatographic column A Phenomenex Gemini C18 110A reverse column(250 mm×4.6 mm ID,5?m),flow rate was 1 mL/min,column temperature was 25 ?,detection wavelength was 332nm and injection volume was 10 ?L.The conditions of mobile phase:the volume ratio of acetonitrile and 1%aqueous acetic acid was 20 to 80 and the condition of flavonoids extract vitexin was 0.2%.3.The effects of flavonoids extract and vitexin were detected by MTT assays:Vitexin concentrations range of 1×10-10 ?g/mL-1×10-1 ?g/mL,effect on osteoblast MC 3T3-E1 role 24 h,48 h and 72 h.With the increase of the dose,the proliferation of obvious effect increased.When the concentration range of 1×10-10?/mL-1×10-6 ?g/mL,their proliferation effect greater than 1×10-5?g/mL-1×10-1 ?g/mL.Drugs on MC 3T3-E1 role 24 h,48 h and 72 h,are capable of varying degrees of promoting the proliferation of MC 3T3-E1 growth.4.In this study,the MC 3T3-E1 cells differentiation research of flavonoids extract vitexin has been researched using colorimetry.The differentiation on MC 3T3-E1 was obviously when the concentration of vitexin and pigeonpea extracts were 1×10-8 ?g/mL and the acting time was 3 d?5 d?7 d,respectively.5.The MTT assay study detected dexamethasone reversed MC 3T3-E1 proliferation effect,vitexin and pigeon pea leaf extract were added after induction processing in cell proliferation rate more than the model group.When vitexin and pigeonpea leaf extracts's concentration are 1×10-8 ?g/mL after 72 h,vitexin group was 37.52 ± 0.34%,the pigeonpea leaf extract was 52.47±2.55%;to some extent vitexin and pigeon pea leaf extract dexamethasone proved have a reversal the role,and have dose-dependent.6.The content of ALP were determined after using vitexin and the mixture extract from the leaf of pigeonpea on cells for 72 h,the concentration of both was 1×10-8 ?g/mL,respectively.And the content of ALP was 17.04±0.54%and 15.06±0.27%,respectively.It explains that dexamethasone could inhibit the differentiation of MC 3T3-E1.7.The von kossa was used to detect MC 3T3-E1 mineralized section to calculate the number of nodules,the results indicate that the same group of cells treated with in number of mineralized nodules increased along with the extension of time in dosing.The number of nodules in the vitexin group was more than that in pigeonpea extract group,when vitexin and pigeonpea leaf extract effect on MC 3T3-E1 for 28 d.It indicated vitexin and pigeonpea extract have the reverse effect to MC 3T3-E1 after induction from dexamethasone.