Primary Evaluation On Cancer Prevention And Treatment Of Dunaliella Salina Extract

Objective:Dunaliella salina (D. salina) is rich in carotene, it has the function of aiti-aging,anticancer,decreasing angiocardiopathy incidence rate,cosmetology and skin protection as well as supplementing vitamin A. At presentβ-carotene which is the main component in D. salina has been the focus of research and development, but other bioactivity compounds except polysaccharides wasn't reported at home and abroad. This study was conducted to investigate the existence of anticancer agents in the D. salina beside polysaccharides andβ-carotene, which would provide a foundation for further development of D. salina.Method:①Preparation of extract:In order to removal the salt of D. salina, washing the D. salina paste with distilled water, drying in the absence of light and grinding to powder. The samples were prepared via petroleum ether,chloroform,ethyl acetate,acetone,ethanol and distilled water extraction respectively. We gained six samples of different polarity. Two samples was obtained from aqueous water which was separated by water extracting and alcohol precipitation technology. Three samples exceptβ-carotene was obtained from Chloroform extract which was separated by column chromatography. Thus, we gained nine samples:A,B2,B3,B4,C,D,E,F1 and F2.②We selected MTT method to evaluate the anticancer effect of the samples on tumor cells proliferation (mouse tumor U14,S180 and human tumor A54,Hela) in vitro and IC50 as the evaluation index.③Lymphocyte proliferation test induced by ConA in vitro was performed to evaluate the immunological activity and its evaluation index is the rate of promoting or inhibiting the lymphocyte.④The attenuation of the samples was evaluated by the protective function test in vitro of the samples towards the bone marrow cells damaged by chemotherapeutics such as E-ADM. And bone marrow cell survival rate was the evaluation index.⑤Based on tumor cells proliferation inhibition test in vitro, the synergistic effect was evaluated by observing the effect of combination of the samples and E-ADM,DDP.⑥Preventing tumor function was evaluated by Ames test. We selected TA98 as tested bacteria and ADM as positive mutagen to observe the anti-mutagenesis of the samples. Result:①131.1gD. salina powder was obtained from 600g D. salina paste washed by distilled water. The extract of yield of A,B (β-carotene and B2,B3,B4),C,D,E and F (F1 and F2) is respectively 5.19%,10.53%,0.92%,0.38%,4.27%,11.67%. Extract B analysised by thin layer chromatography and ultraviolet spectrophotometry containβ-carotene which was gained 146.2mg by silica gel column chromatography.②The first seven samples showed a cytotoxic effect on U14,S180,A549 and Hela kept definite dose-effect relationship. Extract A,B3 exhibited higher cytotoxic effect and water extract has none cytotoxic effect (IC50>1.0mg/mL).③The first seven of extracts from the D. salina including that of extract A,B2,B3,B4,C,D,E may inhibit the proliferation of T lymphocyte of the mouse's spleen induced by ConA in dose manner.④Extract B4 and C may protect the bone marrow cells of mouse from E-ADM. Extract C is more effective than extract B4. Extract C at 2.5,5,10,20,40μg/mL may significantly prevent E-ADM from damaging bone marrow cells and increase the survival rate of marrow cells from 41.71% to 57.62%,55.11%,60.35%,56.71%,57.85%(P<0.05).⑤The result indicated that extract A and B3 combined with DDP on mice U14 show the synergistic effect (Q>1.15). However, the two samples combined wih E-ADM on mice U14 only show the additive effect (Q=0.93～1.05), and on mice S180 show antagonism (Q<0.85).⑥Extract F1 at 0.1,1.0,10.0μg/dish may minimize ADM mutagenesis of TA98 with inhibition rate of 53.38,94.79,66.07%.Low,middle and high dose groups of extract Fl may inhibit the ADM mutagenesis compared with ADM group and there is statistic significance (P<0.05).Conclusion:There are other active agents exceptβ-carotene and polysaccharides in D.salina that are againsr cancer, mutagenesis, increase cytotoxicity of DDP and protect the marrow cells from ADM. Therefore, the D. salina is worth developing as an anticancer drug or food.