Study On Chemical Structures And Bio-activities Of Polysaccharides From Kochia Scoparia

Objective The aim of this study was to filtrate hypoglycemic and immune active site of polysaccharides from kochia scoparia.By modern ways of separation and analysis of clear its structure characteristics and efficacy,preliminary mechanism.To investigate the relationship between the structure of kochia scoparia polysaccharides and its hypoglycemic and immunomodulatory effects.Researching the composition of kochia scoparia polysaccharide which has good pharmacological activity.Methods(1)Extraction and isolation of KSP: Hot water extracting and TCA method was employed to isolate the KSP.The KSP was separated by membrane separation technique into seven parts of relative molecular weight.They were 6 kDa,10 kDa,20 kDa,50 kDa,100 kDa and 150 kDa,respectively.(2)Screening the hypoglycemic activity of crude polysaccharides with different molecular weights: Selecting the alloxan-induced mouse diabetes model and the diabetic rats were treated with different crude polysaccharides.The fasting blood glucose and serum insulin,blood urea nitrogen and glycogen content were measured.Blood glucose activity and screening of active polysaccharide sites.(3)Screening the immunomodulation activity of crude polysaccharides with different molecular weights: Spleen lymphocyte proliferation of mouse and the macrophage phagocytosis were studied.The immunoregulatory activity of polysaccharides was studied and the sites of active polysaccharides were screened.(4)The active polysaccharides were screened by pharmacological experiments and purified by DEAE Cellulose 52 column chromatography to obtain homogeneous polysaccharides.The purity and molecular weight of homogenous polysaccharides were detected by HPGPC-ELSD.The structure of homogeneous polysaccharides were analyzed by high performance liquid chromatography(HPLC),periodate oxidation,Smith degradation,methylation analysis,gas chromatography-mass spectrometer(GC-MS)and infrared spectroscopy(IR).Results(1)Hot water extracting and TCA method was employed to isolate the KSP.The extraction rate was 12.1%.The polysaccharide content was 49.1%.The protein content was 2.7%.The seven parts of different molecular polysaccharides were named respectively KSP-1,KSP-2,KSP-3,KSP-4,KSP-5,KSP-6,KSP-7.prop-ortions of seven parts in total polysaccharides are 4.2%,2.9%,5.2%,28.3%,3.4%,16.1% and 39.9%,respectively.(2)Seven groups of KSP were selected by activity screening about hypoglycemic.After screening,three groups of hypoglycemic activities were obtained,of which the activity of KSP-7 was the best.(3)Seven groups of KSP were selected by proliferation of splenic lymphocytes and peritoneal macrophage phagocytosis of neutral red.After screening,three groups of immunoregulatory activities were obtained,of which the activity of KSP-6 was the best.(4)KSP-6 and KSP-7 were isolated and purified through DEAE-Cellulose 52.After purification,got four homogeneous polysaccharides,named KSP-6a,KSP-6b,KSP-7a and KSP-7b.The molecular weights of four homogenous KSP were 9.11×103 Da,8.44×103 Da,5.58×103 Da and 3.07×103 Da,respectively.KSP-6a was mainly composed of Man,GlcA,GalA,Glc,Gal and Xyl.KSP-6b,KSP-7a and KSP-7b were mainly composed of Man,Rha,GlcA,GalA,Glc,Gal and Xyl.However,their molar ratio were different.The structure of KSP-6a is mainly that main chain was Xyl and Glc.The structure of KSP-6b is mainly that main chain was Xyl and Gal.The structure of KSP-7a is mainly that main chain was Glc and Gal.The structure of KSP-7b is mainly that main chain was Glc and Rha.Conclusions(1)Membrane separation method is suitable for the initial segmentation of polysaccharides.The method can be used to separate the polysaccharides according to different molecular weights.The method can be used for the initial purification,and the process can be further studied and the research efficiency can be improved.(2)kochia scoparia polysaccharides had good hypoglycemic activity and immune activity.(3)KSP-6a and KSP-6b were isolated and purified by KSP-6.The composition of monosaccharide in KSP-6a was the highest in Glc.The composition of monosaccharides in KSP-6b was the highest in Man and Glc.KSP-7a and KSP-7b were isolated and purified by KSP-7 with the best hypoglycemic activity.The composition of monosaccharides was the highest in Glc and Gal.The monosaccharides Ratio and the connection of different,and slightly different composition of monosaccharides.(4)In the case of different monosaccharide composition,with the majority of Man and Glc,immunoregulatory activity is good;to Glc and Gal mostly,hypoglycemic activity is good.