Liver Injury Protective Effection Of Polysacchrides Of Dicliptera Chinensis And The Underlining Mechanism

Hepatic injury is the commom histopathological basis of diverse hepatic diseases. It's continuous existing could trigger the process of hepatic fibrosis, cirrhosis and heptocarcinogenesis. So, controling hepatic injury is the major point in clinical treatment of hepatic diseases. Although the underlying mechanism of hepatic injury is complicated, lipid peroxidation, impaired immunity and dirupted apoptosis are widely accepted.Dicliptera chinensis (L.) Nees. is a trditional herbs of medicine and food, and used in hepatic disease by folks. It's reported that polysaccharides of Dicliptera chinensis (L.) Nees. could decrease sera enzyme in animal hepatic injury mode induced by CCl4 or Bacillus Calmette-Guerin+ lipopolysaccharide. But the comprehensive evalutaion about hepatic injury protection and it's underlying mechanism are not explored. This study focus on the comprehensive hepatic protection effection and underlying mechanism of polysaccharides of Dicliptera chinensis (L.) Nees. in three animal model includes fulminant hepatitis, immunological hepatitis and immunosuppressing model.Objective 1. To investigate the liver protective effects of polysaccharides of Dicliptera chinensis (L.) Nees. on fulminant hepatitis caused by D-galactosamine (D-GalN)/lipopolysaccharide (LPS) in rats. And explore the underlying mechanism.2. To investigate the liver protective effects and it's underlying mechanism of polysaccharides of Dicliptera chinensis (L.) Nees. on acute hepatitis caused by Bacillus Calmette-Guerin (BCG)/lipopolysaccharide (LPS) in mice.3. To investigate the effection of Dicliptera chinensis polysaccharides on immunity in immune suppressed mice induced by cyclophosphamide (CTX).Methods 1. Male Wistar rats were randomly divided into four groups, including normal control group, GalN/LPS-treated group, and two Dicliptera chinensis (L.) Nees. polysaccharides-treated group (150, 300 mg·kg-1, i.g., respectively). The mortality were observed at 6th and 12th hour. Blood were harvested at 6th and 12th hour in survived rats. All blood samples were mesured by ALT,AST,TNF-α,IL-1β. Sample harvested at 12th hour were detected for serum NO additionally. At 12th hour, all survived rats were sacrificed for histopathological evaluation and immunohistochemistry(IHC) assay. Apoptosis associated protein Bcl-2 and Bax were detected by IHC.2. Kunming mice were randomly grouped as normal control group (NC), BCG and LPS treated model group, high dose polysaccharides treated group, medium dose polysaccharides treated group, low dose polysaccharides treated group, biphenyldicarboxylate treated group (postitive control). Activity of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST) were detected. Hepatic hemogenate were used to detect superoxide dismutase (SOD) , malondialdehyde (MDA) and activity of glutathione peroxidase (GSH-PX). HE staining were used to evaluted the pathological change in hepatic tissue.3. Kunming mice were randomly divided into six groups,which includes normal controlgroup, model group, high dose polysacchrides group, mediunm dose polysacchrides, low dose polysacchrides and levamisole treated group. A immunosuppression model were induced by CTX in Kunming mice through subcutaneous injection. Splenic index and thymic index were evaluated by wet weight of spleen and thymus. The monocyte/macrophage phagocytosis were assessed by clearance of ink. Serum hemolysin were detected by colorimetric method. Lymphocytes proliferation rate were assessed by MTT method.Results 1. All animals survived at 6 hours after modeled, pretreatment with polysaccharides of Dicliptera chinensis (L.) Nees. significantly decrease the mortality of the hepatic injuried animals. Pretreatment with polysaccharides of Dicliptera chinensis (L.) Nees. significantly attenuated GalN/LPS-induced severe hepatotoxicity, as evidenced by decreased ALT, AST levels and improved histopathological changes. Polysaccharides of Dicliptera chinensis (L.) Nees. decreased the elevated levels of TNF-α, IL-1βand NO. Polysaccharides of Dicliptera chinensis (L.) Nees. decreased the abnormally elevated expression of Bax, but have no obvious effection on expression of Bcl-2.2. The polysaccharides of Dicliptera chinensis(L.) Nees. could decrease the activity of serum ALT and AST; And decrease MDA, increase SOD and the activity of GSH-PX in hepatic hemogenate. The pathological change were alleviated in hepatic tissue.3. The polysaccharides of Dicliptera chinensis(L.) Nees. could rescue the atrophy of spleen and thymus induced by CTX, speed the clearance of ink, promote the secretion of hemolysin and increase the proliferation rate of splenic lymphocytes in immunity-suppressed mice.Conclusions 1. Dicliptera chinensis (L.) Nees. polysaccharides can provide a definite protective effect against fulminant hepatitis caused by GalN/LPS in rats, which may be mediated by its immunity-regulatory effect to decrease sera level of TNF-α, IL-1βand NO and protection against disrupted apoptosis.2. The polysaccharides of Dicliptera chinensis (L.) Nees. could protect angainst the immunologically hepatic injury in mice, it's underlying mechanism may include anti-oxidative radicals and anti-lipid peroxidation.3. The polysaccharides of Dicliptera chinensis could rescue the immunity function of immune-suppressed mice.