Study On Extraction And Separation Of Total Flavonoids From Pine Needles Of Cedrus Deodara And It’s Anti-tumor Activity In Vitro

Objevtive: To research pinus lambertiana(Cedrus Trew) of Cedrus deodara(Roxb.) Loud. Pine needles of total flavonoids enrichment and purification process by macroporous adsorption resin. Further, using tumour cell experiment in vitro to research its antitumor biological activity and mechanisms.Methods: Using ethanol refluxing extraction method, we get Cedrus deodara(Roxb.) Loud. pine needles total flavonoids. Through static adsorption-desorption experiment, we employ static adsorption-resolution rate and quantity by macroporous adsorption resin on its total flavonoids as filter conditions. Finally, we select the best macroporous adsorption resin from twelve kinds of different polarity,pore diameter and specific surface area. By single factor experiment,we further study dynamic adsorption-desorption optimum conditions, whose may eventually determine the suitable conditions of macroporous adsorption resin purification and enrichment for Cedrus deodara(Roxb.) Loud. pine needles total flavonoids. Moreover, we chose A549,Hep G2,SHG44,MKN45 and He La cells to carry out proliferation inhibition experiment in vitro for obtaining the most sensitive cancer cell on its total flavonoids,which will be used for cell cycle and apoptosis research.Results: We set static adsorption-desorption ratioes and quantity as index,through static tests, which weak polar macroporous resin HPD722 was been get.More, dynamic adsorption-desorption tests has been used to select the best condition of purification total flavonoids of the Cedrus deodara(Roxb.) Loud. pine needles. Finally, the sample mass concentration is 3.85mg/m L, p H 4.0 and 4BV volume. Under 4BV pure water removing impurity,2BV 70% ethanol resolving and three experiment repeating, the purity of total flavonoids has been changed from 14.15% to 54.28% whose purity times and recovery ratio is 3.84 and 89.58% respectively.In the study of vitro proliferation inhibition, the different concentration groups total flavonoids are proliferation inhibition all the tumor cells dose dependent. After 48 hours, the IC50 of human lung A549 and human liver Hep G2 cancer cells are 211.39μ g/m L and 114.12μg/m L respectively. Comparing proliferation inhibition effects of 5 kinds tumor cells in vitro, we find that Hep G2 cells is most sensitive(P<0.05)to Cedrus deodara(Roxb.) Loud. pine needles total flavonoids, in view of this,which cell cycle and apoptosis have been researched by flow cytometry. The result showes that its total flavonoids can block Hep G2 cell cycle in G0/G1 phase and induce apoptosis,that the two ways together play a role of inhibiting tumor cell proliferation in vitro.Conclusion: In order to establish the HPD722 purification and enrichment Cedrus deodara(Roxb.) Loud. pine needles total flavonoids,we use static and dynamic adsorptiondesorption tests through sample liquid concentration, p H, biggest sample amount, removing and elution conditions, different volume fraction of ethanol desorption and desorption volume to research its best dynamic conditions. The proliferation inhibition experiments in vitro showes that different dose groups total flavonoids of Cedrus deodara(Roxb.) Loud. pine needle generate inhibiting effects on 5 tumor cells respectivly. Particularly, its total flavonoids can remarkably(P<0.05) inhibit Hep G2 cell proliferation by retardation it arrests in G0/G1 phase and induces apoptosis.