Study Of Extraction And Purification Of Seabuckthorn Polysaccharide And Their Functions In Vivo And In Vitro

Seabuckthorn polysaccharide is a neutral polysaccharide, composed of Gal, Ara, Man and Glc has a strong scavenging oxygen free radical activity. LPS can stimulate almost all eukaryotic cell morphology, metabolism and gene expression changes, leading to uncontrolled expression of host cell factor-mediated severe infections. As the body’s main detoxification organ, the liver is undoubtedly the most active places clear of LPS, the main mechanism of lipopolysaccharide on liver damage by Kupffer cells and macrophage cytokine production over-activation, so study the function of seabuckthorn polysaccharide on acute liver injury caused by LPS and macrophage cell activation out of control, on the future of the seabuckthorn polysaccharide drugs on hepatitis treatment is great significant.1.Use of water extraction,alcohol precipitation,and gel column chromatography to get pure seabuckthorn polysaccharide.2.1n vitro:Kunming mice were randomly divided into four groups:normal group, model group, seabuckthorn polysaccharide low-dose group, high-dose group intraperitoneal injection of LPS (10mg/kg) to build liver injury model. Pathological observation and ALT, AST detection of pathological changes of liver tissue using RT-QPCR to detect changes in the liver cell factor.As a result, the polysaccharide group, serum ALT, AST and NO levels were significantly lower than the model group, the polysaccharide group liver tissue TNF-a, IL-1β, IL-6, of iNOS mRNA expression was significantly low than the model group, and high doses of polysaccharide effect is moreapparent.3.In vitro:RAW264.7cells were used as experimentalmaterials, and we study the role of seabuckthorn polysaccharide on LPS-stimulated macrophages. We also use PDTC, KT5720, H-7pretreatment of cells to detect the expression of cytokine mRNA by RT-QPCR, and conduct research on the the polysaccharide role of signaling pathways.As a result, the polysaccharides significantly inhibit TNF-α, IL-1β, IL-6and iNOS mRNA expression on LPS-stimulated macrophage. We also found that PDTC and H-7can significantly inhibit the production of cytokines and the role of KT5720is weak.Can be concluded from these studies:seabuckthorn polysaccharide has a significant role in damage caused by LPS,and its mechanism of the role is most likely to participate in the signaling pathways.