Study On The Protective Effects And Mechanism Of Seabuckthorn Polysaccharide On Acute Liver Failure In Mice

This paper studies the protective effects and mechanism of seabuckthorn polysaccharides on mice with acute liver failure in order to provide scientific basis for the hepatoprotective effects of seabuckthorn polysaccharides.Seabuckthorn polysaccharides was extracted by the methods of water extract and alcohol precipitation, and purified by Sephadex gel chromatography. We tested the putity and monosaccharide composition of the purified seabuckthorn polysaccharides. The purity of seabuckthorn polysaccharides was indentified by ultraviolet spectrum scan and the high performance liquid chromatography (HPLC). The results showed that the purified seabuckthorn polysaccharides are a homogeneous material. The composition of monosaccharide of seabuckthorn polysaccharides was determined by HPLC. The results showed that seabuckthorn polysaccharides are mainly composed of5kinds of monosaccharide, Man, Rha, Glu, Gal and Ara. their percentage in the seabuckthorn polysaccharides is12.73%,5.86%,26.71%,6.30%,48.41%, and the mole ratio was2.02:1.02:4.24:1:9.22.Using LPS/D-GalN combined induced acute hepatic failure model to study the hepatoprotective effects of seabuckthorn polysaccharides in mice. One hundred and twenty male C57BL/6mice were randomly divided into six groups:normal control group, model group, dexamethasone positive control group, low, medium and high dose group of seabuckthorn polysaccharides. Mice in the seabuckthorn polysaccharides low, medium and high dose group were gavaged with50,100and200mg·kg-1seabuckthorn polysaccharides for14days. Acute liver injury model was established by intraperitoneal injection of LPS (10μg·kg-1) and D-GalN (700mg·kg-1). The positive control group was treated with intraperitoneal injection of dexamethasone (10mg·kg-1). Serum samples were collected4h after model establishment, and liver tissues were collected8h later. We detected the activity of ALT, AST, SOD, and GSH-PX and MDA content. Liver histological changes were obtained by pathological observation. Serum TNF-α and IL-1β expression was detected by ELISA. The expression of TLR4, p-ERK, NF-κB and PPAR-γ was detected immunohistochemical methods. SOCS3, p-ERK, p-p38MAPK and p-JNK expression in the liver was detected by Western Blotting. The results show that the seabuckthorn polysaccharides significantly inhibited serum ALT and AST activity in mice with acute liver failture (P<0.01, P<0.05).Pretreatment with seabuckthorn polysaccharides promoted the serum and liver SOD and GSH-PX activity (P<0.01, P<0.05), while inhibiting the generation of MDA (P<0.01). Pathological observation showed that the seabuckthorn polysaccharides significantly reduced inflammation caused liver damage. Serum TNF-α and IL-1β expression in the seabuckthorn polysaccharides group also significantly decreased (P<0.01).Furthermore, seabuckthorn polysaccharides inhibited the expression of TLR4, the phosphorylation of ERK, p38MAPK and JNK, and the nuclear expression of the NF-κB p65. Moreove, seabuckthorn polysaccharides didn’t affect the expression of SOCS3and PPAR-γ. Therefore, we speculate that the anti-inflammation and anti-oxidation effects of seabuckthorn polysaccharides are associated with regulation on the TLR4-MAPK-NF-κB signaling pathway, alleviating LPS/D-GalN induced acute liver failure.