Study On The Role And Mechanisms Of Loureirin B On Experimental Hepatic Fibrosis

Partâ… :Loureirin B on experimental hepatic fibrosis and its possible mechanismObjective: To investigate Loureirin B on experimental hepatic fibrosis, and present possible mechanisms, lay the foundation for further study.Method: 60 clean male SD (Sprague-Dawley) rats weighing (200±10) g were randomly divided into 4 groups:Control group, model group, low dose group and high dose group; With thioacetamide (TAA) inducing liver fibrosis model, to observe Loureirin B (the main components of Dragon's Blood) on liver fibrosis in rats body weight,serum transaminase (ALT,AST),fibrosis markers (HA,LN),liver histopathology, hydroxyproline acid (Hydroxyproline, Hyp) changes.Results: To compare with model group, (1) treatment group(including low-dose and high-dose) body weight increases (P<0.05) and liver somatic decreases (P<0.05); (2) liver pathology METAVIR grade, low-dose group and high-dose group were better than the model group; liver fibrosis semi-quantitative score results, the low dose group and high dose group decrease disease activity meter points, but only high dose group inflammation score decreases significantly (P<0.05); fibrosis scoring down low dose group (P＜0.05), high dose group decreases significantly (P＜0.01); (3) The level of high dose group ALT, AST decreases significantly (P＜0.01), while the low dose group ALT, AST decrease significantly (P<0.05); (4) The level of high dose group HA, LN is significantly decreased (P＜0.01), while the low dose group of HA, LN decreased (P＜0.05); (5) low-dose group liver tissue Hyp decreased (P<0.05), Hyp high dose group is significantly decreased (P<0.01).(6) To compare with the control group, model group has significant differences in the indicators (P<0.01).Conclusion:Rat liver fibrosis model induced by TAA is established Successfully;Loureirin B can effectively improve the quality of life in rats with liver fibrosis;Loureirin B can protect TAA-induced liver fibrosis in liver function, reduce inflammation and fibrosis of liver cells, decrease the role of Hyp content;This may be the mechanism of hepatic fibrosis;Loureirin B treatment on liver fibrosis may be positively correlated with the dosage.Part II:Loureirin B on liver fibrosis in rat liver oxidative stress injury in a preliminary studyObjective:To observe Loureirin B on liver fibrosis in rat liver oxidative stress injury and explore the possible mechanism of liver fibrosis.Method:By kit on liver Superoxide dismutase(SOD),Glutathione peroxidase(GSH-Px) activities and Malondialdehyde(MDA) content; Western blotting were detected in rat livera-Smooth muscle actin(a-SMA) expression changes;Reverse transcription polymerase chain reaction (RT-PCR) on liver tumor necrosis factor-a (TNF-a), peroxisome proliferator-activated receptor-a (PPAR-a) mRNA gene expression change.Results:To Compare with model group,(1)High dose group liver tissue SOD, GSH-Px activity than the control group had decreased, but still higher than those of model group (P＜0.05), MDA content was decreased in high dose group (P＜0.05); (2) Western blot showed that the high and low dose treatment group, Dragon's Blood can be reduced by a-SMA expression, but only the high dose group was statistically significant (P＜0.05); (3) in the treatment group were reduced levels of TNF-a mRNA expression, but the only difference between the high dose group was significant (P＜0.05); treatment group were raised PPAR-a mRNA expression, but only high dose group was significant (P＜0.05). (4) Though despite the low dose group showing no significant difference,it reflects the treatment effect may be correlated with the dosage.Conclusion: Liver fibrosis in rats by the intervention of Loureirin B, (1) liver a-SMA expression decreased, suggesting that the activation and proliferation of HSC was inhibited Loureirin B treatment is effective; (2) liver tissue SOD and GSH-Px increased vitality, MDA content is decreased, suggesting Loureirin B can promote the liver, oxygen free radical scavenger, to improve the oxidative stress and reduce oxidative stress injury; (3) liver expression of TNF-a mRNA is significantly reduced and the expression of PPAR-a mRNA level increases, suggesting Loureirin B to weakening the start and progress of liver fibrosis important inflammatory cytokines TNF-a, block liver fibrosis place.Partâ…¢:Loureirin B on liver fibrosis PDGF-BB, TGF-β₁ signaling pathway in rat liver tissueObjective: By testing the changes of platelet-derived growth factor-BB (PDGF-BB), transforming growth factor-β₁ (TGF-β₁) signaling pathway,the possible mechanism of Loureirin B on liver fibrosis.Method:A western blot on liver tissue smad3, smad7, PDGF-BB protein expression changes; RT-PCR method to detect the liver tissue with TGF-β₁, Collage I mRNA expression changes;The change is compared in statistical methods by the analysis software.Results:To compare with model group, (1) high and low dose group may cut smad3, smad7, PDGF-BB protein expression, but the high dose group smad3, smad7 only significant difference in expression levels decreased (P＜0.05), low-dose group has no statistically significant difference; and low-dose group protein levels of PDGF-BB decreased significantly different (P＜0.05), high dose group the expression of PDGF-BB protein is significantly decreased (P<0.01);(2) high and low dose group can be reduced TGF-β₁ mRNA expression (P＜0.05 or 0.01); high and low dose groups were lower Collage I mRNA expression, but only the expression of high-dose group is significantly lower (P＜0.05), while the low dose group is not significant.Conclusion:After the intervention of Loureirin B, (1) Collage I mRNA expression is significantly reduced, suggesting Loureirin B inhibits the synthesis of collagen fibers; (2)The level PDGF-BB protein decreasing suggests Loureirin B may inhibite the proliferation of HSC; (3) TGF-β₁ mRNA expression and smad3 down, but smad7 elevated, suggesting Loureirin B inhibit liver fibrosis may be through TGF-β₁/smad signal transduction pathway.