Extracion, Purfication And Anti-inflammatory Activity Of Corn Bran Arabinoxylan

In this study,Ultrasonic-Microwave assisted alkali used to extract water-unextractable arabinoxylan(WUAX)from corn bran.The optimum preparation conditions were determined by single factor and response surface methodology.The crude WUAX was purified by Cellulose DEAE-52 column and Sephadex G-100 column,respectively.High-performance liquid chromatography,high-performance gel-permeation chromatography,Fourier transform infrared spectral analysis,scanning electron microscopy(SEM)and NMR analytical techniques were used to compare the characterization of the main eluent components.The RAW 264.7 macrophages induced by lipopolysaccharide was to study the anti-inflammatory activity of AX-1-1?AX-2-1 and AX-3-1.The main results were as follows:1.Optimization and purification of WUAX form corn bran by Ultrasonic-Microwave assisted alkali processIn this study,Ultrasonic-Microwave assisted alkali was applied to extract WUAX from the corn bran.Based on the single factor and response test,the four factors of ultrasonic power,concentration of sodium hydroxide,solvent-to-material ratio and ultrasonic-microwave synergetic time were optimized.The results showed that the maximum WUAX yield of 27.78 ±0.17% was obtained with ultrasonic power 500 W,sodium hydroxide 0.30 mol/L,solvent-to-material ratio 30 mL/g,ultrasonic-microwave synergetic time 25 min.The preliminarily purified were using by Cellulose DEAE-52 column.There was one peak of water-eluted arabinoxylan(AX-3),and two peaks of salt-eluted arabinoxylans(AX-1 and AX-2).Each main fraction was further purified on a Sephadex G-100 column with distilled water to obtain high-purity arabinoxylans.Three major eluent components were collected and named AX-1-1,AX-2-1 and AX-3-1 for structural elucidation and anti-inflammatory activity evaluation.2.Characterization of AX-1-1,AX-2-1 and AX-3-1 from corn branFT-IR analysis processed the arabinoxylan characteristic peaks of AX-1-1?AX-2-1 and AX-3-1;Scanning electron microscopy(SEM)showed that the three main eluent fractions were mainly aggregated,thin and smooth.High-performance gel-permeation chromatography showed that AX-1-1,AX-2-1 and AX-3-1 were homogenous polysaccharides with molecular weights of 287,305 and 280 KDa,respectively.High-performance liquid chromatography showed that the main monosaccharide components of AX-1-1,AX-2-1 and AX-3-1 were arabinose and xylose with A/X ratios of 0.78,1.02 and 0.96,respectively.The structure of the three purified arabinoxylans was analyzed by H-NMR and C-NMR spectra.It was determined that AX-1-1,AX-2-1 and AX-3-1 mainly consisted of a main chain linked by ?-D-xylose,and ?-L-arabinofuranose formed a side chain structure at the position of C(O)-2 or C(O)-3,which had similar chemical composition and spatial structure.3.Anti-inflammatory Activity of AX-1-1,AX-2-1 and AX-3-1 from corn branAnti-inflammatory effects of AX-1-1,AX-2-1 and AX-3-1 were evaluated using RAW 264.7 macrophages stimulated by lipopolysaccharide(LPS).AX-3-1 showed significant anti-inflammatory activity,which not only reduced the secretion of inflammatory cytokines,including tumor necrosis factor(TNF-?),Interleukin-1?(IL-1?)and Interleukin-1?(IL-1?),but also reduced the level of Nitric Oxide(NO)release.AX-3-1 inhibited the expressions of Cyclooxygenase-2(COX-2)and inducible nitric oxide synthase(iNOS),and inhibited the nuclear factor-?B(NF-?B)activation of lipopolysaccharide(LPS)stimulated RAW 264.7 macrophages.