Polygonum Taipaishanease Kung Polysaccharides Sulfation And Pharmacodynamic Studies Of Modificated Produts

Chinese medicine polysaccharide hashas an important immune regulation, anti-oxidation, anti-tumor, anti-viral, anti-cancer, anti-glycation, anti-inflammatory and other immune pharmacological activity, And toxic side effects, no residue, no drug resistance. In recent years, studies have shown that Chinese medicine polysaccharide after sulfated modification to enhance their pharmacological activity. In this experiment, Polygonum taipaishanease Kung as materials, Polygonum taipaishanease Kung polysaccharide(PTKP) was extracted by Cordiofolia, PTKP sulfated modification by chlorosulfonic acid- pyridine method, The orthogonal experiment to modify conditions were optimized, Barium sulfate- gelatin turbidity method was used to determine the degree of substitution(DS), Fourier infrared spectroscopy measuring its structure changes, swine testis cells(ST cell) as a model, CCK 8 method and real-time fluorescent quantitative PCR method to detect PTKP, sPTKP inhibitory effect of TGEV on ST cell; By in vitro antibacterial test for detection of porcine endogenous E. coli and Shigella inhibition of sPTKP,Determination of the impact of sPTKP on pig peripheral blood lymphocyte factor IL- 2 and TNF-α by ELISA. To determine the sulfated Polygonum taipaishanease Kung polysaccharide in vitro antiviral and antibacterial activity, etc.Following results were obtained: 1 Optimal conditions PTKP sulfated modified is chlorosulfonic acid: pyridine ratio of 1:10, the reaction temperature is 80 ℃, the reaction time was 1h, sPTKP sulfate group substitution degree of 1.57. 2 The maximum non-toxic concentration PTKP of 20μg / mL, the maximum non-toxic concentration sPTKP of 80μg / mL. In the same concentration of PTKP and s PTKP, three kinds of modes of the Virus infection After Admadministration is best..In the same mode of administration, sPTK, PTKP of TGEV inhibition rate proportional to its concentration, respectively;The same concentration, the same mode of administration, sPTK of TGEV inhibition rate higher than PTKP. PTKP, sPTKP by three kinds of administration could significantly reduce the level of transcription of mRNA relative expression levels of TGEV, These PTKP, sPTKP adsorption resistance of TGEV is best. At same administration, the relative expression of TGEV mRNA is inversely proportional to the concentration of PTKP and sPTK. The same administration,and concentration, sPTK group Inhibition of TGEV is significantly higher than PTKP group(P <0.01). 3 The PTKP minimum bactericidal concentration of porcine endogenous Shigella and E. coli minimum bactericidal concentrations were 20 mg / mL and 40 mg / mL;the sPTKP minimum bactericidal concentration of porcine endogenous Shigella and E. coli minimum bactericidal concentrations were 10 mg / mL and 20 mg / mL. 4 In concentrations within safe limits, sPTKP, PTKP treated, the content of IL-2 and TNF-α level were significantly higher than the control group of normal cells, and IL-2, TNF-α content and sPTKP, PTKP concentrations positively correlated. Under the same concentration, cells treated by sPTKP group, the content of IL- 2 and TNF- α significantly higher than that of PTKP group. Conclusion: Optimal conditions PTKP sulfated modified is chlorosulfonic acid: pyridine ratio of 1:10, the reaction temperature is 80 ℃, the reaction time was 1h, In vitro tests confirmed thatsulfated modified can significantly improve the PTKP activity of anti-TGEV and Inhibition of porcine endogenous Shigella and E. Coli ability, PTKP vitro Lymphocyte Immune factor IL-2 and TNF-α secretion activity has also been significantly enhanced.