Paeoniflorin Attenuates Carbontetrachloride Induced Acute Liver Injury And Liver Fibrosis

1.Paeoniflorin Attenuates Carbon Tetrachloride-Induced Acute Liver InjuryObjective:The present study aims to investigate the roles and specific mechanisms of paeoniflorin on carbon tetrachloride-induced acute liver injury.Methods:Mice were randomly divided into six groups(n=6): normal control group,paeoniflorin control group,carbon tetrachloride model group and three doses of paeoniflorin intervention groups.Acute liver injury was induced by intraperitoneal injection of carbon tetrachloride(5 μl/g,dissolved in olive oil).Mice received 10,30,100 mg/kg paeoniflorin every 8 hour a time for three times before carbon tetrachloride administration.24 hours after carbon tetrachloride administration,mice were anesthetized and killed,and then serum and liver tissues were collected to further analysis.The activities of alanine aminotransferase and aspartate aminotransferase were determined to evaluate hepatocellular damage.Tissue sections were stained with hematoxylin and eosin staining to examine histopathological changes.The activity of caspase-3 in liver tissue was measured to evaluate hepatocyte apoptosis.The activities of SOD,GSH-PX and CAT and the content of MDA and GSH in liver tissues were detected using kits following manufacturer’s directions.Serum of TNF-α and IL-6 levels were detected by enzyme-linked immunosorbent assay.Quantitative reverse transcription polymerase chain reaction was used to detect the m RNA expression of HO-1 in liver tissue.Results:When compared with control group,carbon tetrachloride administration increase serum alanine aminotransferase and aspartate aminotransferase levels.Carbon tetrachloride induced hepatocyte apoptosis.Hepatic MDA was markedly increased,GSH-PX,CAT,GSH and SOD were significantly decreased in the carbon tetrachloride exposure model group.Carbon tetrachloride significantly increased TNF-α and IL-6 level in serum.When compared with carbon tetrachloride model group,pretreatment paeoniflorin obviously reduce serum alanine aminotransferase and aspartate aminotransferase levels.Pretreatment paeoniflorin could restrain hepatocyte apoptosis.Pretreatment paeoniflorin obviously inhibited MDA content,increased GSH content,enhanced GSH-PX,CAT and SOD activities in the liver of mice.Pretreatment paeoniflorin significantly decreased TNF-α and IL-6 level in serum.Hepatic HO-1 m RNA was markedly up-regulated by paeoniflorin treatment.Conclusion:Paeoniflorin could effectively prevent carbon tetrachloride-induced acute liver injury by reducing hepatocyte apoptosis,inhibiting lipid peroxidation and pro-inflammatory cytokines production,promoting antioxidant protein HO-1 expression.2.Paeoniflorin Attenuates Carbon tetrachloride-Induced Liver FibrosisObjective:The present study aims to investigate the roles and specific mechanisms of paeoniflorin on carbon tetrachloride-induced liver fibrosis.Methods:Mice were randomly divided into four groups(n=6): normal control group,paeoniflorin group,carbon tetrachloride model group and paeoniflorin intervention groups.Mice were administered with carbon tetrachloride(5 μl/g,i.p.,dissolved in olive oil)for seven weeks to induce liver fibrosis and concomitantly treated with paeoniflorin(100 mg kg-1 day-1)for last six weeks.Mice were anesthetized and killed,and then serum and liver tissues were collected to further analysis.The activities of alanine aminotransferase and aspartate aminotransferase were determined to evaluate hepatocellular damage.Tissue sections were stained with hematoxylin and eosin staining to examine histopathological changes.Masson staining,Sirius staining and the content of hydroxyproline in liver tissue was measured to evaluate fibrosis.The activities of SOD,GSH-PX and CAT and the content of MDA and GSH in liver tissues were detected using kits.Hepatic of TNF-α and IL-6 levels were detected by enzyme-linked immunosorbent assay.Quantitative reverse transcription polymerase chain reactionwas used to detect the m RNA expression of α-SMA,desmin,vimentin,collagenⅠand HO-1 in liver tissue.Western blot was used to detect theprotein expression of α-SMA,desmin,vimentin,collagen Ⅰ and HO-1 in liver tissue.HO-1 activity was measured by bilirubin generation.Results:When compared with control group,carbon tetrachloride increase serum alanine aminotransferase and aspartate aminotransferase levels.Carbon tetrachloride induced hepatic fibrosis.Carbon tetrachloride significantly increasedthe m RNA and protein expression of α-SMA,desmin,vimentin and collagen Ⅰ in liver tissue.Hepatic MDA was markedly increased,GSH-PX,CAT,GSH and SOD were significantly decreased in the carbon tetrachloride exposure model group.Carbon tetrachloride significantly increased TNF-α and IL-6 level in serum.When compared with carbon tetrachloride group,pretreatment paeoniflorin could obviously reduce serum alanine aminotransferase and aspartate aminotransferase levels.Pretreatment paeoniflorin could restrain liver fibrosis.Paeoniflorin significantly decreasedthe m RNA and protein expression of α-SMA,desmin,vimentin and collagenⅠ in carbon tetrachloride-induced liver fibrosis.Pretreatment paeoniflorin obviously inhibited MDA content,increased GSH content,enhanced GSH-PX,CAT and SOD activities in the liver of mice.Pretreatment paeoniflorin significantly decreased TNF-α and IL-6 level in serum.Hepatic HO-1 m RNA and protein expression and activity was markedly up-regulated by paeoniflorin treatment.HO-1 activity was increased by paeoniflorin pretreatment.Conclusion:Paeoniflorin could effectively prevent carbon tetrachloride-induced liver fibrosis by reducing hepatic fibrosis,inhibiting lipid peroxidation and pro-inflammatory cytokines production,promoting antioxidant protein HO-1 expression and activity.