Study On Extraction Of Adlay Bran Oil And Its Anti-tumor Effect On A549 And RD Cells In Vitro

Adlay bran is an abundant byproduct of coix seed.Its output is about 40 thousand tons every year in Guizhou province.Adlay bran contains more than 20%(w/w)oil,in which more than 80% are polyunsaturated fatty acids,and it also contains a variety of nutrients such as squalene,vitamin E,oryzanol and so on.The oil extracted from coix seed has antitumor activity and has been applied in the clinical treatment of cancer patients.It has been found that the methanol / ethanol extracts from other parts of the adlay also have some anti-tumor and anti-inflammatory functions,but there is little research on whether the adlay bran oil has anti-tumor effect.At present,adlay bran is often treated directly as livestock feed,resulting in the waste of highquality resources.In order to improve the exploitation and utilization of adlay bran,the research on the extraction of adlay bran oil,the analysis of the oil quality and its effect of antitumor in vitro were had been systematically studied in this paper.(1)The influence of different extraction methods(aqueous extraction,aqueous enzymatic extraction and aqueous ethanol extraction)on free oil yield was studied.Results showed: the extraction rate of aqueous ethanol extraction was the highest.The effect of pulverization method on the particle size and distribution of oil and protein in adlay bran were studied.Based on this concrete process,the conditions of aqueous ethanol extraction were optimized.The optimal conditions of particle size: 30 μm;the optimal conditions of aqueous ethanol extraction: extraction pH 7.0,ethanol concentration 30%(v/v),solid-to-liquid ration 1:5(w/v),extraction temperature 50°C,extraction time 2 h.Under above optimal conditions,the free oil yield reached 71.44%±0.82%,the residual oil content of sendiment phase was 22.72%±0.74%,the residual oil content of emulsion phase was 2.98%±0.43%,the residual oil content of water phase was 2.86%±0.15%.(2)The quality of adlay bran oil extracted by aqueous ethanol was analysed.The results showed that the physical and chemical properties of crude oil had no significant difference from other extraction methods.The acid value and peroxide value of crude oil extracted by aqueous ethanol extraction were both lower than that of the organic solvent extraction.The adlay bran oil obtained by aqueous ethanol extraction contained 10 fatty acid components,and the composition and content of fatty acids were basically the same as those of coix seed oil.The similarity of the adlay bran oil and coix seed oil was more than 0.95 by fingerprint and similarity evaluation system,indicating the consistency of the main chemical components.In the inhibition experiments of fatty acid synthetase(FASN)in vitro,the inhibitory effect of adlay bran oil on FASN was higher than that of coix seed oil,reaching 84.6%.It indicated that adlay bran oil could inhibit the tumor by inhibiting the activity of fatty acid synthetase.(3)We studied the anti-tumor effect of adlay bran oil in vitro.The results showed that the adlay bran oil had significant inhibitory effect on the proliferation of A549 and RD tumor cells,and it could significantly increase the rate of early apoptosis and late apoptosis of tumor cells.It could also obviously increase the expression of IL-10,IFN-β and Atg 5 immune factors in cells,thereby reducing the degree of inflammatory reaction.There was no significant difference between the adlay bran oil with three antitumor drugs(KangLaiTe,docetaxel and paclitaxel)in the effect of anti-tumor and promoting the expression of cellular immune factors.The results of transcriptional analysis showed that the total number of differentially expressed genes was 4090 in RD tumor after treated with adlay bran oil.GO enrichment and KEGG enrichment showed that the differentially expressed genes mainly enriched in multiple biological processes and signaling pathways closely related to tumor,such as cell circulation,DNA replication,carbon metabolism,and so on.It was indicated that adlay bran oil had broad prospects for development and utilization in anti-tumor treatment.