Studies On Thepurification Of Fucoidan Extracted From Kjellmaniella Crassifolia Cultured In Dalian And Its Anti-inflammatory Activity

Fucoidan is a water-soluble sulfuric acid heteropolysaccharide extracted from brown algae and has various biological activities.In this paper,the fucoidan sulfate from Kjellmaniella crassifolia cultured in Dalian was used as raw material,and the two components F1 and F2 were separated and purified by DEAE-Sepharose Fast Flow ion exchange chromatography.The recovery rates of these two components were 23.25% and 25.19%,respectively.The sulfate group contents of F1 and F2 were 2.94% and 41.34%,respectively,and the total sugar content was 75.00% and 44.91%,respectively.The monosaccharide composition was analyzed by high performance liquid chromatography,F1 mainly contained mannose,and F2 mainly contained fucose.Using Fourier transform infrared spectroscopy,the F1 sulfate group was distributed at the C2 and C3 positions,and the F2 sulfate group was distributed at the fucose C4 position.The results of animal experiments showed that all dose groups of DF,F1 and F2 decreased the levels of IL-1?,IL-6 and TNF-?,and increased the level of IL-10.Compared with the model group,the low and high dose groups of DF and F1,high dose group of F2 highly significantly reduced the levels of IL-6 and TNF-?,the low and high dose groups of F1 and high dose group of F2 highly significantly reduced IL-1? level and all dose groups of F1 significantly increased IL-10 level.The high dose group of F2 had the best effect on the reduction of IL-1? and IL-6.Meanwhile,compared with the model group,the high dose group of DF,all dose groups of F1 and F2 highly significantly reduced the activity of MDA,and the low dose group of DF significantly reduced the activity of MDA,and the high dose group of F2 showed best effect.In adition,all dose groups of DF,F1 and F2 significantly increased the activity of SOD and GSH-Px compared with the model group.The high dose group of F1 was the most effective in the activity of SOD and GSH-Px.The mRNA expressions of inflammation-related genes COX-2,iNOS,I?B-? and NF-?B were detected by RT-PCR.Compared with model group,all dose groups of DF,the high dose groups of F1 and the low dose group of F2 significantly reduced the expression of COX-2 mRNA.For the expression level of iNOS mRNA,the high dose groups of F1,all dose groups of F2 reduced iNOS mRNA highly significantly(p?0.01)while the low dose groups of DF and the low dose groups of F1 reduced iNOS mRNA significantly(p?0.05).Obviously,high dose group of F1 had best effect.Compared with model group,the low and high dose groups of DF,F1 and F2 highly significantly increased the expression of I?B-? mRNA(p?0.01).For the expression of NF-?B mRNA,the high dose groups of DF and F1,all dose groups of F2 highly significantly reduced NF-?B mRNA levels(p?0.01),the low dose groups of DF and F1 significantly decreased the expression of NF-?B mRNA(p?0.05).Also the results showed that high-dose group of F1 had best effect for inhibiting inflammatory by adjusting the expression of NF-?B mRNA.Therefore fucoidan sulfate from Kjellmaniella crassifolia cultured in Dalian could regulate the inflammation by adjusting the inflammatory genes.