Research On Intestinal Flora Imbalance And Probiotics With Alcholic Liver Disease

Objective To investigate the role of intestinal flora imbalance and probiotics and TGF-β/Smad signal pathway in the progress of alcohol-induced liver fibrosis in mice.To investigate the character of bowel flora and the theropeutic role of probiotics in alcoholic liver disease(ALD)patient.To explore the theoretical basis for potential treatments of liver fibrosis.Methods Part ? A total of 36 C57 mice were randomly and equally divided into 3 different dietary regimes: Group I(alcohol injury;received alcohol);group II(alcohol injury with flora imbalance;received alcohol plus lincomycin hydrochloride)and group III(alcohol injury with corrected flora imbalance;received alcohol,lincomycin hydrochloride and extra probiotics).To evaluate the degree of liver fibrosis and liver biochemical indicators in mice,half the mice in each group were sacrificed through cervical dislocation to harvest liver tissues for pathologic assessments and collect blood samples from the retro orbital plexus;HSCs were addi-tionally isolated from the remaining mice.The levels of ALT,AST and ALP in the mice sera were detected with ALT,AST or ALP kits respectly.HE staining of liver tissue sections was performed for Pathologic assessments.Masson staining of liver tissue sections was performed to evaluate the degree of liver fibrosis.The pathological assessments of livers were performed according to AID histology score standard.Annexin V fluorescein isothio-cyanate/propidium iodide double staining and flow cytometry was also performed to detect the apoptosis of HSCs.The expression of mothers against decapentaplegic homolog(smad)3 and smad4 proteins in hepatic stellate cells(HSCs)of the mice was examined using western blot anaysis.Part П 65 adult males admitted with a diagnosis of ALD were enrolled in this randomized,clinical study in 2weeks.Patients were randomized to receive 14 days of probiotics plus standard thropy versus and standard thropy(abstinence,high caloric,protein,low fat diet,vitamins,etc)only.Stool cultures,liver enzymes and endoxemia were performed before the trail and were repeated after the therapy.Results were compared between groups and with 30 healthy,matched controls who did not consume alcohol.Results Part ? Mice in group II exhibited significantly higher serum ALT,ALP and AST levels(P<0.05)compared with those in group I and group III.Serum ALT,ALP and AST levels of the mice in group III were significantly lower compared with that of the mice in group II(P<0.05).According to the standard of ALD scores,the scores of hepatocyte ballooning,liver steatosis,liver inflammation and fibrosis are summarized in the stydy,which shows that the degree of liver damage was significantly higher in the mice of group II compared with the mice in group.However,these effects were limited following the correction of intestinal flora imbalance in the mice of group III(P<0.05).Apoptosis in HSCs was significantly inhibited in the livers of the mice in group II as compared with that in group I(P<0.05).The present study also observed that correcting the intestinal flora imbalance in the mice of group III significantly increased apoptosis in HSCs compared with that in group I(P<0.05).The expression levels of smad3 and smad4 were significantly upregulated in HSCs of the mice in group II compared with that in group I(P<0.01).By contrast,the expression levels of smad3 and smad4 were significantly downregulated in HSCs of the mice in group III compared to that in the mice of group II(P<0.01).Part П Compared to healthy controls,ALD patients had significantly reduced numbers of bifidobacteria and enterococci.Liver enzymes were significantly elevated in the alcoholic group compared to the healthy control group,indicating that these patients did have significant alcohol-induced liver injury.The level of endoxemia was significantly higher than the control.After 14 days of therapy of probiotics,ALD patients had significantly increased quantity of bifidobacteria,lactobacilli and enterococci compared to the standard therapy group.Despite similar values at study initiation,patients treated with probiotics had significantly lower AST and ALT activity(P<0.05)and significantly decreased of endoxemia(P<0.05)at the end of treatment than those treated with standard therapy alone.In a subgroup of 35 subjects with diagnosis alcoholic hepatitis,therapy of probiotics was associated with a significant reduction in liver enzymes.In this subgroup,there was a significant end of treatment mean AST(P<0.05)and endoxemia(P<0.05)reduction in the probiotics therapy versus the standard therapy group.Conclusions Intestinal flora imbalance can affect alcohol-induced liver fibrosis in C57 mice by aggravating the progress of liver fibrosis.While,The present study demonstrated that correcting intestinal flora imbalance is necessary to break down the progress.Subsequent to correcting intestinal flora imbalance,the proliferation and activation of HSCs was attenuated,which reduced excessive production of extracellular matrixes by downregulating the TGFβ/smad signaling pathway.ALD patients have significantly altered bowel flora compared to healthy controls.Short-term oral supplementation with probiotics was associated with restoration of the bowel flora and greater improvement in alcohol-induced liver injury than standard therapy alone.Therefore,correcting intestinal flora imbalance is crucial and can be an effective method in the treatment of liver fibrosis.