| Sparganium stoloniferum is a herbaceous plant,which has the functions of promoting blood circulation and relieving pain.In this paper,the crude polysaccharides were extracted step by step from Sparganium stoloniferum with hot water,chelating agent,1mol/L Na OH and 4 mol/L Na OH buffer yielded four different polysaccharide fractions RSB,RSC,RSDA and RSCA.Seven fractions,RSB-1,RSB-2,RSB-3,RSB-4,RSDA-1,RSDA-2 and RSDA-3,were isolated from RSB and RSDA by DEAE-Cellulose ion exchange chromatography.The structure of polysaccharides was identified by chemical and instrumental analysis methods including GC,GC-MS,NMR and partial acid hydrolysis.The biological activity of polysaccharide was studied by pharmacological experiments.These results would be provide a scientific theoretical basis for the developmentt of Sparganium stoloniferum.The main results of this paper are as follows.(1)The results of physicochemical properties showed that the contents of total sugar of RSB,RSC,RSDA and RSCA were 54.51%,70.01%,85.36%and 88.34%,respectively.Uronic acid contents of RSB,RSC,RSDA and RSCA were 5.43%,3.16%,0.59%and 0.38%,respectively.Protein contents of RSB,RSC,RSDA and RSCA were24.00%,19.92%and 0.95%,of which no protein was detected in RSCA.Microscopic and thermal stability analysis showed that the microstructures of RSB,RSC,RSDA and RSCA were significantly different,of which RSDA and RSCA had relatively high thermal stability.The results of in vitro anti-oxidation and immunomodulatory activities showed that RSB and RSDA had relatively high biological activities.(2)The chemical composition analysis results showed that the composition of the seven fragments was significantly different.RSB-4,RSDA-2 and RSDA-3 were identified as homogeneous components by HPGPC,and the average molecular weight of this three fractions were 9.32×10~5,8.23×10~5and 8.18×10~5Da,respectively.The monosaccharide composition analysis showed that RSDA-2 mainly composed of glucose,arabinose,xylose,and galactose at the molar ratio of 1.00:5.83:9.01:1.37;RSDA-3 mainly composed of arabinose and xylose at the molar ratio of 1.00:1.50;RSB-4 was glucan.(3)The results of structural analysis showed that the backbone of RSB-4 might be consisted of(1→3)-α-D-Glcp,in which the branched chains T-β-D-Glcp and(1→6)-α-D-Glcp were substituted at the O-6 position of(1→3)-α-D-Glcp;The backbone of RSDA-2 might be consisted of(1→2)-β-D-Xylp,(1→3)-β-D-Xylp and(1→3)-α-D-Arap,in which the branched chains T-α-L-Xylf,T-β-D-Glcp,T-α-D-Galp,(1→4)-β-L-Arap and(1→4)-β-D-Xylp were substituted at the O-4 position of the main chains;The backbone of RSDA-3 might be consisted of(1→2)-β-D-Xylp,(1→3)-β-D-Xylp and(1→3)-α-D-Arap,in which the branched chains T-α-L-Xylf,and(1→4)-β-D-Xylp were substituted at the O-4 position of the main chains.(4)RSB-4,RSDA-2 and RSDA-3 can effectively promote the proliferation of macrophages,with the highest proliferation index were 1.46,1.49 and 1.45,respectively.RSB-4,RSDA-2,and RSDA-3 had effects on the production of NO induced by LPS.When the dose concentration was 20μg/m L,the highest inhibition rates were 23.74%,25.89%and 22.21%,respectively.The loss of branch showed an effect on anti-inflammatory activities of cells.(5)The mechanism of the anti-inflammatory polysaccharide RSDA-2 was analyzed by RT-PCR and Western Blot experiments.RT-PCR results showed that RSDA-2 can significantly inhibit the expression of inflammatory factors i NOS,IL-1βand TNF-α,thereby slowing and preventing the progress of inflammatory response.The results of Western Blot experiments showed that RSDA-2 can inhibit the activation of NF-κB pathway of macrophages by inhibiting the phosphorylation of IκB and p65 induced by LPS,thereby inhibiting the expression and secretion of inflammatory mediators and exerting anti-inflammatory effects. |
PDF Full Text Request