The Study Of Misgurnus Anguillicaudatus Lyophilized Powder On Hepatoprotective And Anti-hepatic Fibrosis Effects

Objective: To observe the hepatoprotective and anti-hepatic fibrosis effects ofMisgurnus anguillicaudatus lyophilized powder, MLP) and to explore its possibletherapeutic mechanism.Methods:（1） The LD50or the maxiumu dose of MLP was determined with mouseacute toxicity test. The mouse body weight and toxicity reaction were recorded andobserved for successive seven days when MLP was administrated for the highest dose of12.48g/kg.（2） In CCl₄-induced liver injury mouse, three parts of MLP, full MLP, muscleMLP and skin MLP, were administrated to screen the effective part. Mouse were treatedwith MLP for10days, and received intraperitoneal injection of0.4％CCl₄solution in11thd. The hepatoprotective effect was evaluated by analysis of serum ALT and AST levels.（3）In CCl₄-induced acute liver injury mouse, three doses of MLP （100,200and400mg/kg）were administrated to detect their therapeutic effect by analyzing serum ALT and ASTlevels.（4） In CCl₄-induced hepatic fibrosis mouse，the effect of MLP （50,100,200mg/kg） was evaluated by measuring liver function tests and observing histopathologyresult, including ALT, AST contents in serum and Hyp content in tissue.（5） InDMN-induced hepatic fibrosis rats, indicators in common with hepatic fibrosis mouse wereanalyzed to evaluate therapeutic action of MLP treatments.（6） On the base of MLPanti-hepatic fibrosis experiment, to deeply explore its anti-fibrosis mechanism, MMPs,TIMP-1, HA, LN and collagen PC-III, C-IV, TGF-β1contents were tested by ELISAmethod, and α-SMA, TIMP-1protein contents and distribution were detected usingWestern blot and immunohistochemistry methods.Results:（1） MLP was safe within the dose of12.48g/kg, because the mouse treatedwith maximum MLP had no symptoms of poisoning and death.（2） In the experiment ofscreening effective part of MLP, musle MLP and full MLP treatments can significantlyreduce mice serum ALT and AST levels （P＜0.05）, in contrast skin MLP hadn’t obvious action. Full MLP was choosed as administration part taking into account the productionprocess.（3） In CCl₄-induced acute liver injury mouse，the serum ALT and AST levelswere significantly decreased compared with model group （P＜0.05）, especiallyhigher-dose group （P＜0.01）.（4） In CCl₄-induced hepatic fibrosis mouse, the Hypcontent in tissue and ALT/AST level in serum were evidently decreased in MLP treatedgroups. In CCl₄-induced hepatic fibrosis mice, MLP treated groups can evidently decreaseHyp content in tissue and ALT/AST level in serum. HE and Masson staining results in themodel group showed that the normal liver lobule structure was damaged, and the liver cellwere presented ballooning-like degeneration or feather-like degeneration, accompany witha lot of inflammatory cell infiltration in liver portal. Collagen fiber volume was abundantlyincreased and even complete fake hepatic lobule was formed. However, the degree of liverdamage and fibrosis were ameliorated by MLP. At the same time, SSS scores in MLPtreatment groups were declined compared with before treatment＞2cents. These resultssuggested that MLP can significantly reduce CCl₄-induced hepatic fibrosis.（5） InDMN-induced hepatic fibrosis rats, the serum ALT and AST levels and Hyp content inliver tissuewere significantly decreased in MLP groups （P＜0.01, P＜0.05）, but serumAlbumin content （P＜0.05） was increased. Hepathological results showed that the livercells were almost resumed normal, inflammatory cells and collagen fibers weresignificantly reduced after MLP administrated.（6） The ELISA detection results indicatedthat the levels of HA, LN, PC-III, C-IV were reduced, the MMP-2level was up-regulatedand the MMP-9level was down-regulated, TGF-β1protein expression wasreduced.Western and immunohistochemical results displayed that TIMP-1, α-SMA.protein expression were reduced.Conclusion: The results suggest that MLP has hepatoprotective and anti-hepaticfibrosis effects and its mechanism of anti-hepatic fibrosis may be associated with reducingECM synthesis, advancing ECM degradation, inhibiting HSC activation, inhibitingcytokine TGF-β1expression.