Research advances in natural drug and food monomers in prevention and treatment of alcoholic liver injury / 临床肝胆病杂志

A large number of data show that the prevalence rate of alcoholic liver injury （ALI） is increasing year by year， and it has become one of the main causes of death due to chronic liver diseases such as liver cancer and liver cirrhosis. Quitting drinking is the main method for the prevention of ALI in modern medicine， and the main treatment methods include Western medicine with antioxidant and anti－fibrotic effects and nutritional support. However， Western medicine tends to have an unsatisfactory treatment effect and can only alleviate initial symptoms， and severe ALI still requires surgical treatment. Studies have shown that the monomers extracted from natural drugs and foods have obvious preventive and therapeutic effects on ALI， with high safety and easy access. Therefore， this article systematically summarizes the main natural drug and food monomers used for the prevention and treatment of ALI and proposes the idea of the combination of drug and food for the prevention and treatment of ALI from the perspective of paying attention to the whole process of health， in order to explore more effective prevention， health care， and treatment methods and provide ideas for research on the prevention and control of ALI.

Effect of Shenling Baizhusan on Alcoholic Liver Injury in Rats by Regulating Ferroptosis Based on Nrf2 Signaling Pathway / 中国实验方剂学杂志

ObjectiveTo observe the effect of Shenling Baizhusan on the intervention of the nuclear factor erythroid 2-related factor 2 （Nrf2） signaling pathway by regulating ferroptosis in rats with alcoholic liver injury. MethodForty SD rats were randomly divided into model group， polyene phosphatidylcholine group， and high， medium， and low-dose Shenling Baizhusan groups， with 8 rats in each group. Another 8 SD rats were taken as blank group. The model group， polyene phosphatidylcholine group， high， medium， and low-dose Shenling Baizhusan groups were given 10 mL·kg-1 liquor by gavage for modeling， and the blank group was given equal volume of distilled water by gavage. After 4 h of daily alcoholic administration， 143.64 mg·kg-1 of polyene phosphatidylcholine group was given to the polyene phosphatidylcholine group， 15， 7.5， 3.75 mg·kg-1 of Shenling Baizhusan were given to Shenling Baizhusan high， medium， and low-dose groups， respectively， and the blank group and the model group were given equal volume of distilled water. The gavage lasted for 6 weeks. The levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST）， glutamyl transpeptidase （GGT）， total cholesterol （TC）， and triglyceride （TG） were detected by automatic biochemical analyzer. The levels of tumor necrosis factor-α （TNF-α） and interleukin-β （IL-β） were detected by the enzyme-linked immunosorbent assay （ELISA）. The levels of lipopolysaccharide （LPS）， malondialdehyde （MDA）， superoxide dismutase （SOD）， glutathione （GSH）， and Fe+ were detected by biochemical assay. The pathological changes in the liver were observed by hematoxylin-eosin （HE） staining and oil red O staining. The mRNA expression levels of Nrf2， heme oxygenase-1 （HO-1）， glutathione peroxidase 4 （GPX4）， ferritin heavy polypeptide 1 （FTH1）， and nuclear factor-κB （NF-κB） were detected by Real-time polymerase chain reaction （Real-time PCR）. The protein expression levels of Nrf2， HO-1， GPX4， FTH1， p65， and phosphorylation （p）-p65 were detected by Western blot. ResultAs compared with the blank group， the levels of liver function （ALT， AST， and GGT） and blood lipids （TC and TG） in the model group were significantly increased （P<0.05）. The liver showed obvious steatosis， with a large number of fat deposition， the oxidative stress and inflammatory factors were significantly increased （P<0.05）， and the level of Fe+ was significantly increased in model group （P<0.05）. The protein expression levels of Nrf2， HO-1， GPX4， and FTH1 was significantly down-regulated （P<0.05）， and those of p65 and p-p65 was significantly up-regulated in the model group （P<0.05）. The mRNA expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly down-regulated （P<0.05）， and the mRNA expression level of NF-κB was significantly up-regulated （P<0.05）. As compared with the model group， the levels of liver function （ALT， AST， and GGT） and blood lipids （TC and TG） in the high-dose and medium-dose Shenling Baizhusan groups were significantly decreased （P<0.05）， liver steatosis was significantly improved， fat deposition was significantly reduced， oxidative stress and inflammatory factors were significantly decreased （P<0.05 ）， and Fe+ level was significantly decreased （P<0.05）. In the high-dose and medium-dose Shenling Baizhusan， the protein expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly up-regulated （P<0.05）， and those of p65， p-p65 were significantly down-regulated （P<0.05）. The mRNA expression levels of Nrf2， HO-1， GPX4， and FTH1 were significantly up-regulated （P<0.05）， and the mRNA expression level of NF-κB was significantly down-regulated （P<0.05）. ConclusionShenling Baizhusan can effectively reduce liver injury in rats with ALD， regulate steatosis and fat deposition， and play an antioxidant and anti-inflammatory role in the liver. Its mechanism may be related to the inhibition of ferroptosis in hepatocytes by up-regulating the Nrf2 signaling pathway to improve oxidative stress

Lipid nanoparticle delivery of siRNA targeting Cyp2e1 gene attenuates subacute alcoholic liver injury in mice / 浙江大学学报·医学版

OBJECTIVES@#To investigate the effect and mechanism of lipid nanoparticle (LNP) delivery of small interfering RNA (siRNA) targeting Cyp2e1 gene on subacute alcoholic liver injury in mice.@*METHODS@#siRNA targeting Cyp2e1 gene was encapsulated in LNP (si-Cyp2e1 LNP) by microfluidic technique and the resulting LNPs were characterized. The optimal dose of si-Cyp2e1 LNP administration was screened. Forty female C57BL/6N mice were randomly divided into blank control group, model control group, si-Cyp2e1 LNP group, LNP control group and metadoxine group. The subacute alcoholic liver injury mouse model was induced by ethanol feeding for 10 d plus ethanol gavage for the last 3 d. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, and the superoxide dismutase (SOD) activity as well as malondialdehyde, reactive oxygen species, glutathione, triacylglycerol, total cholesterol contents in liver tissue were measured in each group, and liver index was calculated. The expression of genes related to oxidative stress, lipid synthesis and inflammation in each group of mice were measured by realtime RT-PCR.@*RESULTS@#Compared with the model control group, the levels of liver index, serum ALT, AST activities, malondialdehyde, reactive oxygen species, triacylglycerol, total cholesterol contents in liver tissue decreased, but the SOD activity as well as glutathione increased in the si-Cyp2e1 LNP group (all P<0.01). Hematoxylin-eosin staining result showed disorganized hepatocytes with sparse cytoplasm and a large number of fat vacuoles and necrosis in the model control group, while the si-Cyp2e1 LNP group had uniformly sized and arranged hepatocytes with normal liver tissue morphology and structure. Oil red O staining result showed si-Cyp2e1 LNP group had lower fat content of the liver compared to the model control group (P<0.01), and no fat droplets accumulated. Anti-F4/80 monoclonal antibody fluorescence immunohistochemistry showed that the si-Cyp2e1 LNP group had lower cumulative optical density values compared to the model control group (P<0.01) and no significant inflammatory reaction. Compared with the model control group, the expression of catalytic genes P47phox, P67phox and Gp91phox were reduced (all P<0.01), while the expression of the antioxidant enzyme genes Sod1, Gsh-rd and Gsh-px were increased (all P<0.01). The mRNA expression of the lipid metabolism genes Pgc-1α and Cpt1 were increased (all P<0.01) and the lipid synthesis-related genes Srebp1c, Acc and Fasn were decreased (all P<0.01); the expression of liver inflammation-related genes Tgf-β, Tnf-α and Il-6 were decreased (all P<0.01).@*CONCLUSIONS@#The si-Cyp2e1 LNP may attenuate subacute alcoholic liver injury in mice mainly by reducing reactive oxygen levels, increasing antioxidant activity, blocking oxidative stress pathways and reducing ethanol-induced steatosis and inflammation.

Protective effects of Lonicerae Japonicae Flos against acute alcoholic liver injury in rats based on network pharmacology / 中国中药杂志

This study aims to explore underlying mechanism of Lonicerae Japonicae Flos(LJF) in protecting rats against acute alcoholic liver injury(ALI) based on mitogen-activated protein kinase(MAPK) pathway. First, the targets of LJF in preventing ALI were predicted by network pharmacology and the component-target-pathway network was constructed, so that the key targets of LJF components acting on MAPK pathway were screened. Second, male SD rats were randomized into the control(KB) group, model(MX) group, positive(YX) group, and LJF high-(GJ), medium-(ZJ), and low-(DJ) dose groups. Each administration group was given(ig) corresponding drugs for 7 days and KB group and MX group received(ig) equal volume of distilled water every day. Except for KB group, rats were given Chinese spirit(56%, 3 days) for ALI modeling. The levels of aspartate transaminase(AST), alanine transaminase(ALT), interleukin-6(IL6) and tumor necrosis factor-α(TNF-α) in serum and malondialdehyde(MDA), glutathione(GSH), superoxide dismutase(SOD) and glutathione peroxidase(GSH-Px) in liver tissue of rats in each group were detected. Furthermore, we employed quantitative real-time PCR(qRT-PCR) to probe the effects of LJF on the key targets of MAPK pathway in ALI rats. A total of 28 active components of LJF were screened from TCMSP database, and 317 intersected with ALI-related targets. According to Kyoto encyclopedia of genes and genomes(KEGG) pathway enrichment analysis, the 317 targets involved 226 pathways, which were mainly liver disease, inflammation, immunity, apoptosis and other related pathways. According to the MAPK pathway-target-active component network, the key active components of LJF, such as chlorogenic acid, hederagenol, and hyperoside, acted on 25 key targets of MAPK pathway. The results of in vivo experiments showed decreased levels of AST, ALT, and MDA in DJ, ZJ, and GJ groups(P<0.01 or P<0.05), reduced levels of IL6 in DJ and GJ groups(P<0.01 or P<0.05), and improved levels of SOD and GSH in ZJ and GJ groups(P<0.01 or P<0.05). The results of qRT-PCR demonstrated that the expression levels of mitogen-activated protein kinase kinase 4(MAPK2 K4) and mitogen-activated protein kinase 3(MAPK3) were decreased in DJ, ZJ, and GJ groups(P<0.01). The network pharmacology and experimental verification showed that the active components in LJF can reduce the inflammatory factor level and enhance the activities of SOD and GSH-Px by inhibiting the expression of key targets of MAPK pathway, thus alleviating and preventing liver damage caused by alcohol.

Protective effect and mechanism of Wangshi Baochi Pills against acute alcoholic liver/stomach injury in mice / 中国中药杂志

As a common disease worldwide, alcoholic liver injury is caused by long-term or excessive intake of alcohol and triggers cell death due to alcohol metabolism and reactive oxygen species(ROS)-mediated cytotoxicity. Wangshi Baochi(WSBC) Pills have been widely adopted in clinical practice for evacuating stasis, resolving turbidity, clearing heat, tranquilizing mind, invigorating sto-mach, promoting digestion, purging fire and removing toxin. This study aimed to investigate the efficacy of WSBC Pills in dispelling the effect of alcohol and protecting against acute alcoholic liver/stomach injury in mice, and preliminarily investigate its possible mole-cular mechanism. The results found that the preventive treatment with WSBC Pills contributed to elevating the activity of alcohol dehydrogenase(ADH) and its expression in liver and shortening the time required for sobering up of mice with acute alcoholic liver injury. The staining of liver pathological sections as well as the detection of serum aspartate aminotransferase(AST) and alanine aminotransferase(ALT) and liver ROS levels revealed that WSBC Pills protected the liver by reducing serum AST and ALT. It suppressed oxidative stress-induced liver injury by lowering liver ROS and elevating superoxide dismutase(SOD), and the liver-protecting effect was superior to that of silibinin. Western blot assay confirmed that WSBC Pills inhibited the oxidative stress by up-regulating SOD1 and NAD(P)H: quinone oxidoreductase 1(NQO-1). In addition, WSBC Pills lowered the ROS level to protect against the acute alcoholic stomach injury in mice. The findings have suggested that WSBC Pills alleviated the acute alcoholic liver/stomach injury in mice by increasing ADH and resisting oxidative stress.

Study on the mechanism of Flos Puerariae and Semen Hoveniae in treatment of alcoholic liver injury based on network pharmacology and molecular docking / 浙江大学学报·医学版

OBJECTIVE@#To explore the mechanism of Flos Puerariae and Semen Hoveniae in treatment of alcoholic liver injury (ALI) based on network pharmacology and molecular docking.@*METHODS@#The information of chemical constituents and targets of Flos Puerariae and Semen Hoveniae was collected from TCMSP and Swiss databases, and the threshold values of oral bioavailability (OB) ≥ 30%, drug likeness (DL) ≥0.18 were used to screen the potential active compounds. The GeneCard and DrugBank databases were used to obtain the targets corresponding to ALI. The common targets were queried using Venn Diagram, and the network of PPI and Gene Ontology (GO) functional enrichment analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were performed through DAVID and Reactome database. Autodock Vina software was used for molecular docking of potential ingredients and key targets.@*RESULTS@#A total of 21 potential active compounds and 431 therapeutic targets were gathered in Flos Puerariae and Semen Hoveniae, which involved 273 biological functions, 90 KEGG pathways and 362 Reactome pathways. The GO functions involved protein binding, ATP binding, etc.; the KEGG pathways mainly included PI3K-Akt signaling pathway and TNF signaling pathway; the Reactome pathways contained signal transduction and immune system, etc. The results of molecular docking showed that 21 potential active ingredients had good affinity with the core targets Akt1, TP53 and IL-6.@*CONCLUSIONS@#The network pharmacology and molecular docking analysis demonstrate the synergetic effect of Flos Puerariae and Semen Hoveniae with multi-compounds, multi-targets and multi-pathways in the treatment of ALI; and also predict the possible medicinal substance, key targets and pathways, which provides clues for the new drug development and mechanism research.

Protective Effect of Ficus pandurata Extract on Alcohol-induced Acute Liver Injury Base on Pyroptosis / 中国实验方剂学杂志

Objective:To study the protective effect of Ficus pandurata extract on acute alcoholic liver injury based on pyroptosis mechanism.Method:The 56 male C57BL/6 mice were randomly divided into normal control group， model control group， positive control group（60 mg·kg-1）， fresh medicine water extract group（48 g·kg-1）， dry drug water extract group（48 g·kg-1），dry drug 50% alcohol extract group（48 g·kg-1） and dry drug 95% alcohol extract group （48 g·kg-1）， 8 mice in each group.Positive control and different solvent extract groups of Ficus tenuifolia were intragastrically administrated for 18 days，once a day，while normal group and model group were given the same volume of pure water intragastrically. After 15 days of continuous gavage， mice received 50％ ethanol（12 mL·kg-1）intragastrically for 3 days to induce acute alcoholic liver injury model except for the normal control group. At 14 h after the last treatment，serum and liver samples were obtained，the serum content of alanine aminotransferase （ALT） and aspartate transaminase（AST） were determined， the histopathologic changes of the hepatic tissues were observed by hematoxylin ecosin（HE） staining.The content of malondialdehyde (MDA) in liver was determined by thiobarbituric acid (TBA) and the content of lactate dehydrogenase (LDH) was determined by microplate method. Western blot and TUNEL assay kit was used to detect the cell pyroptosis rate.Result:Compared with normal group， ALT， AST， MDA and LDH levels in the model group were significantly increased， liver index was significantly increased，TUNEL staining positive， inflammatory factors and pyroptosis related protein expressions were significantly increased （P<0.05）. Compared with model control group， the ALT，AST ，MDA and LDH of the drug intervention group decreased significantly （P<0.05）. The liver index decreased in different degrees， and the expression of inflammatory factors and pyroptosis related protein in the water extract treatment group decreased significantly （P<0.05）.Conclusion:The root extract of Ficus pandurata Hance has protective effect on acute alcoholic liver injury， and the mechanism of water extract might relate to inhibiting hepatocyte pyroptosis.

Mechanism of Anti-chronic Alcoholic Liver Injury in Rats of Tibetan Medicine Lagotis brachystachys Extracts by TLR2/MyD88/NF-κB and NALP3 Signaling Pathway / 中国实验方剂学杂志

Objective::To established the model of chronic alcoholic liver injury in rats by long-term(8 weeks) alcoholic gavage, to study the effects of Tibetan medicine Lagotis brachystachys extracts on Toll-like receptor(TLR)2/myeloid differentiation factor 88(MyD88)/nuclear factor kappa B (NF-κB)and NOD like receptor protein 3(NALP3) signaling pathways and study preliminary the mechanism of action of chronic alcoholic liver injury. Method::Sixty male Sprague-Dawley rats were randomly divided into normal group, model group, bifendate positive drug group (0.1 g·kg-1) and L. brachystachys low, medium and high-dose groups (0.5, 1, 2 g·kg-1), the corresponding drugs were given at 10 mL·kg-1 in each morning, and the 56 degree Liquor was administered by the afternoon gradient alcoholic gavage method.After 8 weeks, the levels of serum aspartate transaminase (AST), serum alanineaminotransfease(ALT), serum total cholesterol(TC), triglyceride(TG), interleukin-1β(IL-1β), and the liver levels of L-glutathione(GSH)were measured. The expression of TLR2, MyD88, NF-κB and NALP3 protein in liver were detected by Western blot.Hematoxylin-eosin (HE) staining was used to observe the pathological changes of liver tissue. Result::Compared with normal group, the serum levels of AST, ALT, TC, TG and IL-1β in model group were significantly increased (P<0.05, P<0.01). Compared with model group, the serum AST, ALT, TC, TG and IL-1β levels were decreased in the various doses of L. brachystachys, and the high dose group was particularly effective (P<0.05, P<0.01). Compared with normal group, the GSH level in the liver homogenate of model group decreased significantly, and the difference was not statistically significant. The levels of TLR2, MyD88, NF-κB and NALP3 in the liver tissue of model group were significantly increased (P<0.05, P<0.01). The GSH levels in the liver and the protein expression of TLR2, MyD88, NF-κB and NALP3 were decreased in L. brachystachys group (P<0.05, P<0.01). The liver pathological section showed that L. brachystachys can improve the pathological changes of rat liver tissue. Conclusion::L. brachystachys can protect liver from alcohol-induced chronic liver injury in rats. The mechanism was related to TLR2/MyD88/NF-κB and NALP3 signaling pathway.

Protective effect of kinsenoside on acute alcohol-induced liver injury in mice

Abstract Anoectochilus roxburghii (Wall.) Lindl., Orchidaceae, is a Chinese medicinal plant which can be effective for some diseases such as hepatitis, nephritis, pneumonia. Its active ingredient is kinsenoside. The mechanisms of kinsenoside on the liver-protective effect have not been fully explored until today. The present study was aimed to investigate the protective effect and mechanism of kinsenoside on acute alcoholic liver injury. The protected activity of kinsenoside (10, 20 and 40 mg/kg) were investigated on acute alcoholic liver injury in mice. Male C57BL/6 J mice were fed with non-fat feed for 30 days and oral administrated 14 ml/kg bw of ethanol (50%) on the 31st day. The activities of serum aspartate aminotransferase, serum alanine aminotransferase, triacylglyceride and very low density lipoprotein were determined in serum. The hepatic levels of oxidative stress as glutathione, malondialdehyde were measured in liver homogenates. The levels of cytochrome P450 2E1 (CYP2E1) were measured by immunohistochemistry. Furthermore, histopathological observations were carried out on the separated livers of mice. It was suggested that the trends of acute hepatic injury and fatty degeneration induced by alcohol were reduced in the ethanol group after kinsenoside treatment. Compared to ethanol groups, triacylglyceride, malondialdehyde, very low density lipoprotein, reduced glutathione, serum alanine aminotransferase and serum aspartate aminotransferase levels of kinsenoside (20, 40 mg/kg) groups were decreased (p < 0.05). Meanwhile kinsenoside significantly decreased the level of protein CYP2E1. In conclusion, kinsenoside enhances antioxidant capacity of mice and antagonizes alcohol-induced lipid metabolism disorders. Besides, kinsenoside inhibits alcohol-caused hepatocyte apoptosis, reduces oxidative stress, and relieves hepatocyte death, which may be a mechanism of kinsenoside in the treatment of alcoholic liver.

Protective effect of Schisandra chinensis total lignans on acute alcoholic-induced liver injury related to inhibiting CYP2E1 activation and activating the Nrf2/ARE signaling pathway

ABSTRACT Schisandra chinensis (Turcz.) Baill., Schisandraceae, is a well-known traditional Chinese medicine used mainly as a recipe for hepatoprotection. Modern researches have revealed that the hepatoprotection is related to its lignans and crude polysaccharide. In this study, we examined the effect and mechanism of S. chinensis total lignans on the liver injury induced by alcoholic. S. chinensis total lignans were extracted with ethanol extraction. The liver index, alanine aminotransferase and aspartate aminotransferase levels in serum of the rat culture supernatant were examined. The malondialdehyde level and superoxide dismutase activities in serum and liver tissue, and triacylglyceride content in liver tissue were tested. Western blot was conducted to determine cytochrome P450 2E1 expression in liver tissue of rats. The results showed that S. chinensis total lignans administration significantly inhibited alcohol-induced liver injury. In exploring the underlying mechanisms of S. chinensis total lignans action, we found that it significantly decreased Alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (AKP), Glutamyl transpeptidase (γ-GT), reactive oxygen species (ROS) and malondialdehyde (MDA) level in livers/serum and inhibited the gene expression level of CYP2E1 in rat livers. The Nuclear factor erythroid-2 related factor 2 (Nrf2) gene expression and Nuclear factor erythroid-2 related factor 2 (Nrf2) protein nuclear transfer increased significantly, and significantly increased the expression of downstream target gene and protein heme oxygenase-1 (HO-1), Glutamate--cysteine ligase regulatory subunit (GCLM), NAD(P)H:quinine oxidoreductase 1 (NQO1). Moreover, S. chinensis total lignans decreased production of pro-inflammatory markers including Tumor Necrosis Factor-α (TNF-α), Interleukin-1beta (IL-1β) and Interleukin-6 (IL-6). In conclusion, these results suggested that the inhibition of alcohol-induced liver injury by S. chinensis total lignans is associated with its ability to inhibiting CYP2E1 activation and activating the Nrf2/Antioxidant response element(ARE) signaling pathway.

Comparison of contents and effects of lignans from 4 strains of Schisandra chinensis / 吉林大学学报(医学版)

Objective: To campare the differences of contents and relative effects of lignans from 4 strains of Schisandra chinensis, and to provide the experimental basis for the precise selection and application. Methods: The contents of 7 kinds of lignans from 4 strains of Schisandra chinensis were determined by high performance liquid chromatography (HPLC). The antioxidant effects of 4 strains of Schisandra chinensis were compared by measuring the abilities of reduction, scavenging of DPPH free radical and inhibiting linoleic acid oxidation in vitro Sixty male ICR mice were randomly divided into normal control group, alcoholic liver injury model group (model group), Schisandra chinensis YH group, (YH group), Schisandra chinensis SH group (SH group), Schisandra chinensis 156-5-2 group 156-5-2 group) and Schisandra chinensis 50-1-5 group (50-1-5 group), 10 mice in each group. One hour after the last administration, the mice in normal control group were given the equal volume of distilled water, and those in the other groups were intragastrically given 50% alcohol 12 mL • k g- 1 ) . All the mice were fasted, but with free access to water for 12 h and executed, and the liver and serum were obtained. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in serum of the mice, and the superoxide dismutase (SOD) activity, the levels of triglyceride (TG) and malondialdehyde (MDA) in the liver tissue of the mice were determined. Results: The total content of lignan monomers in Schisandra chinensis YH was the highest, the second was Schisandra chinensis SH, and 156-5-2 and 50-1-5 were later. Except for 156-5-2, other 3 kinds of Schisandra chinensis had strong antioxidant effects, and the order of antioxidant activity was Y H > S H > 5 0 - l - 5 . Compared with model group, the ALT and AST levels in the serum and the levels of MDA and TG in the liver tissue of the mice in YH group were decreased significantly (P 0. 05); the ALT level in the serum of the mice in 156-5-2 group was decreased (P 0 . 05). Conclusion: In 4 strains of Schisandra chinensis, YH has the highest content of lignans, has the strongest antioxidant effect in vitro, and has the best protective effect on acute alcoholic liver injury.

Gypenosides protect acute alcoholic liver injury through Nrf2/nf-kb signaling pathway in mice / 中国药理学通报

Aim To investigate the protective effect of gypenosides on acute alcoholic liver injury in mice and its mechanism. Methods The BALB/c mice were divided into five groups, control, alcoholic injury group and three protection groups by different doses of gypenosides. The mice in protection groups were given different doses of gypenosides everyday for one week. After the mice were sacrificed, the peripheral blood and the liver were collected. The pathology of liver tissue sections was observed by HE staining. The ALT, AST, TC and TG were determined by automatic biochemical analyzer. TNF-a and IL-6 mRNA levels were determined by real-time PGR. The content of MDA and GSH, the activities of SOD and CAT and the level of GSH were determined by kits respectively. The protein levels of proteins of Nrf2 and NF-kB signaling pathways were assayed by Western blot. Results Alcohol treatment increased the levels of ALT, AST, TC and TG as well as the expression levels of TNF-a and IL-6. At the same time, alcohol treatment increased the content of MDA and decreased the activities of SOD, CAT and GSH. The alcohol treatment also inhibited the Nrf2 signaling pathway and induced the NF-kB translocation to nuclear. The gypenoside supplementation significantly inhibited all the above changes in a dose-dependent way. Conclusion The gypenoside supplementation shows significant protection on the acute alcoholic liver oxidative injury in mice by regulating Nrf2/NF- kB signaling pathway.

Protective effect of immunomodulating peptide(PGPIPN) derived from beta-casomorphin in bovine milk on acute alcohol-induced liver injury / 中国药理学通报

Aim To investigate the protective effect of immunomodulating peptide(PGPIPN) on the acute al-coholic liver injury in mice. Methods Kunming mice were randomly divided into control group, model group,glutataione(GSH) group, PGPIPN low dose group, PGPIPN moderate and high dose groups. The mice were treated with different doses of PGPIPN or GSH for two weeks except control group and model group. The acute alcoholic liver injury model was in-duced by gavage with 56° alcohol for three days. The indices including the activities of AST,ALT in serum, and the contents of TNF-α, MDA, SOD and GSH-Px in liver were examined. Liver histopathological changes were examined by HE staining. Results Compared with control group,the levels of serum ALT,AST and the contents of TNF-α, MDA significantly increased, while the contents of SOD and GSH-Px significantly de-creased in model group. There was hepatocyte apopto-sis and inflammatory cell infiltration in liver tissues. Compared with model group, the activities of serum ALT, AST and the contents of TNF-α, MDA were re-markably reduced in PGPIPN high dose group. The contents of SOD and GSH-Px significantly increased in PGPIPN high dose group. PGPIPN could alleviate the injury of liver. Conclusion PGPIPN has certain pro-tective effect on acute alcoholic liver injury of mice, providing a theoretical guidance.

Hepaprotective Effect of Standardized Ecklonia stolonifera Formulation on CCl₄-Induced Liver Injury in Sprague-Dawley Rats

The liver is an essential organ for the detoxification of exogenous xenobiotics, drugs and toxic substances. The incidence rate of non-alcoholic liver injury increases due to dietary habit change and drug use increase. Our previous study demonstrated that Ecklonia stolonifera (ES) formulation has hepatoprotective effect against alcohol-induced liver injury in rat and tacrine-induced hepatotoxicity in HepG2 cells. This present study was designated to elucidate hepatoprotective effects of ES formulation against carbon tetrachloride (CCl₄)-induced liver injury in Sprague Dawley rat. Sixty rats were randomly divided into six groups. The rats were treated orally with ES formulation and silymarin (served as positive control, only 100 mg/kg/day) at a dose of 50, 100, or 200 mg/kg/day for 21 days. Seven days after treatment, liver injury was induced by intraperitoneal injection of CCl₄ (1.5 ml/kg, twice a week for 14 days). The administration of CCl₄ exhibited significant elevation of hepatic enzymes (like AST and ALT), and decrease of antioxidant related enzymes (superoxide dismutase, glutathione peroxidase and catalase) and glutathione. Then, it leaded to DNA damages (8-oxo-2′-deoxyguanosine) and lipid peroxidation (malondialdehyde). Administration of ES formulation inhibited imbalance of above factors compared to CCl₄ induced rat in a dose dependent manner. Real time PCR analysis indicates that CYP2E1 was upregulated in CCl₄ induced rat. However, increased gene expression was compromised by ES formulation treatment. These findings suggests that ES formulation could protect hepatotoxicity caused by CCl₄ via two pathways: elevation of antioxidant enzymes and normalization of CYP2E1 enzyme.

extract (Wuzhi Tablet) protects against chronic-binge and acute alcohol-induced liver injury by regulating the NRF2-ARE pathway in mice

Alcohol abuse leads to alcoholic liver disease and no effective therapy is currently available. Wuzhi Tablet (WZ), a preparation of extract fromthat is a traditional hepato-protective herb, exerted a significant protective effect against acetaminophen-induced liver injury in our recent studies, but whether WZ can alleviate alcohol-induced toxicity remains unclear. This study aimed to investigate the contribution of WZ to alcohol-induced liver injury by using chronic-binge and acute models of alcohol feeding. The activities of ALT and AST in serum were assessed as well as the level of GSH and the activity of SOD in the liver. The expression of CYP2E1 and proteins in the NRF2-ARE signaling pathway including NRF2, GCLC, GCLM, HO-1 were measured, and the effect of WZ on NRF2 transcriptional activity was determined. We found that both models resulted in liver steatosis accompanied by increased transaminase activities, but that liver injury was significantly attenuated by WZ. WZ administration also inhibited CYP2E1 expression induced by alcohol, and elevated the level of GSH and the activity of SOD in the liver. Moreover, the NRF2-ARE signaling pathway was activated by WZ and the target genes were all upregulated. Furthermore, WZ significantly activated NRF2 transcriptional activity. Collectively, our study demonstrates that WZ protected against alcohol-induced liver injury by reducing oxidative stress and improving antioxidant defense, possibly by activating the NRF2-ARE pathway.

Effects of Pinus Yunnanensis on Acute Alcoholic Liver Injury in Mice / 中国中医药信息杂志

Objective To study the effect of Pinus yunnanensis on acute alcoholic liver injury in rats and explore its mechanism. Methods A model of acute alcoholic liver injury in mice was prepared by alcohol. The mice were randomly divided into normal control group, model group, positive control group, Pinus yunnanensis low-, medium-and high-dose groups. Mice in the medicine group were given the corresponding medicine by gavage once a day for 7 days. After the last three hours of intragastric administration, the liver and spleen index, ALT, AST and GSH in serum, SOD, MDA and NO in liver homogenates were measured. Histopathological changes of liver were observed by HE staining. Results Compared with model group, Pinecone of Pinus unnanensis high-, medium- and low-dose groups could significantly reduce the liver index in mice (P0.05). HE staining results showed that, the damage of liver tissue in mice of Pinus yunnanensis was significantly improved compared with the model group. Conclusion Pinus yunnanensis has protective effects on acute alcoholic liver injury in mice.

"The Effect of""Gut-liver Axis""on the Development of Alcoholic Liver Disease and the Intervention of Linderae Radix" / 浙江中医药大学学报

[Objective] To analyze relations between alcoholic liver disease(ALD) and gut-liver axis, and the intervention effects of Linderae radix(LR) on both of them. [Methods] Research reports about ALD,gut-liver axisand LR on anti ALD at home and abroad in recent years were reviewed, and effects ofgut-liver axisin the occurrence and development of ALD and LR on ALD treatment were summarized. [Results] Recent studies show that there is an abnormalgut-liver axisin ALD,gut permeability and intestinal endotoxemia(IETM) induced by excessive alcohol consumption is one of the key pathogenic factors for the occurrence and development of ALD. Improving intestinal functions to alleviate IETM would be an efficient way to treat ALD. Linderae radix (LR) is a traditional Chinese medicine commonly used for Qi-regulating in clinic ,it possesses a good effect for gastrointestinal function regulation and as reported in recent researches,it has shown a good protective effect on ALD by improving the abnormal gut-liver axisand alleviating IETM. [Conclusions] The abnormalgut-liver axisinduced by excessive alcohol intake is the key factor for ALD formation and development. LR possesses a protective function on ALD viagut-liver axisimprovement.

Protective effect of Zhi Zi Da Huang Tang on rats with alcoholic fatty liver / 吉林大学学报(医学版)

Objective:To establish the alcoholic fatty liver(AFL) animal models,and to explore the protective effect of Zhi Zi Da Huang Tang (ZZDHT) on the rats with AFL and its dosage.Methods:A total of 54 SD rats were divided into normal control group (n=10) and model control group (n=44).The rats in model control group were given alcohol by lavage (50% ethanol solution 6.0 mL·kg-1) combined with high-fat feed to establish the rat models of AFL.After 4 weeks,the rats in model control group were randomly divided into model group(treated with water),simvastatin group (10 mg·kg-1),low and high doses (5.0 and 10.0 g·kg-1) of ZZDHT groups,and there were 10 rats in each group.Every 2 weeks during the process,the body weights and levels of serum total cholesterol (TC),triglyceride (TG),aspertate aminotransferase (AST),and alanine aminotransferase (ALT) were detected.After 4 weeks,the body weights and liver weights of the rats were detected;the levels of TC,TG,AST,ALT,alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in serum and liver tissue of the rats of were detected;the levels of tumor necrosis factor-alpha (TNF-α),interleukin-6 (IL-6) and interleukin-1β(IL-1β) in serum of the rats were detected;the morphology of liver tissue was observed by HE staining and the pathological examination was performed.Results:Compared with normal control group,the levels of serum TC,AST,ALT,TG,TNF-α,IL-6,and IL-1β of the rats in model group 4 weeks after administration were significantly increased (P<0.05 or P<0.01).Compared with model group,the levels of serum TC,TG,AST,and ALT of the rats in low and high doses of ZZDHT groups 4 weeks after administration were decreased (P<0.05 or P<0.01);the levels of serum TNF-α and IL-6,IL-1β of the rats in high dose of ZZDHT group were decreased (P<0.01);the levels of serum TNF-α and IL-1β of the rats in low dose of ZZDHT group were decreased (P<0.01);the levels of serum ADH and ALDH in liver tissue of the rats in low and high doses of ZZDHT groups and simvastatin group were decreased(P<0.05 or P<0.01).The HE staining result showed that compared with model group,the pathological conditions of the liver tissue of the AFL rats in ZZDHT groups were significantly improved.Conclusion:ZZDHT can significantly improve the liver injury caused by high fat diet combined with alcohol and fat liver lesions.

Impacts of Quercetin on Alcoholic Liver Injury in Rats / 医学研究杂志

Objective To study the impacts of quercetin on alcohol-stimulated liver injury in rats.Methods Thirty Sprague-Dawley rats were randomly divided into control,model,quercetin treatment (40mg/kg,80mg/kg,160mg/kg) groups.Ethanol plus 0.5ml fish oil were used to induce alcoholic liver injury for 6 weeks.Liver injury was evaluated using pathological examination and serum ALT levels.The plasma endotoxin and serum TNF-α,IL-1 and IL-18 levels were analyzed by ELISA method.The expression of CD14,LBP,TNF-α,IL-1 and IL-18 proteins in the liver were measured by Western blot.Results The increased serum ALT,fatty degeneration,focal necrosis and inflammatory cell infiltration in the liver were investigated in model group.In addition,plasma endotoxin,serum TNF-α,IL-1,IL-18 levels,and the expression levels of CD14,LBP,TNF-α,IL-1,and IL-18 proteins significantly elevated in model group compared with control group (P ＜ 0.05).However,quercetin treatment improved histological changes,and significantly reduced the levels of plasma endotoxin and serum TNF-α,IL-1,IL-18,as well as the expression of CD14,LBP,TNF-α,IL-1 and IL-18 proteins (P all ＜ 0.05).Fatty degeneration was still investigated in quercetin treatment group,but focal necrosis and inflammatory cell infiltration disappeared.Conclusion Quercetin can protect liver against necrosis and inflammation induced by alcohol,and the mechanism may involve its effect on the reduction of plasma endotoxia,and inhibition of Kupffer cell activity and proinflammatory cytokine expression.

Protective effects of Liuwei Wuling tablet on acute alcoholic-induced liver injury in mice / 中国中药杂志

The protective action and the relevant mechanism of Liuwei Wuling tablet on acute alcoholic hepatic injury in mice were investigated. All the C57BL/6 mice were divided randomly into 7 groups including blank, model, bifendate (150 mg•kg⁻¹, positive control) and experimental groups consisted of extremely low dose (0.1 g•kg⁻¹), low (0.5 g•kg⁻¹), upper (4 g•kg⁻¹) and high dose (8 g•kg⁻¹) of Liuwei Wuling tablet groups. The acute liver injury model was induced by modified method that the model, positive control and experimental groups were orally administrated 56% alcohol (6 g•kg⁻¹) twice at 12 hour intervals on the fifth day after drugs administration. After 12 hours, the mice were sacrificed to contribute blood and liver for biochemical and histological examinations. Compared with the model, the activities of ALT and AST in serum decreased significantly in different Liuwei Wuling tablet groups. Meanwhile, in liver tissue, the levels of TG, MDA, TNF-α and IL-1β reduced obviously while the GSH and SOD activities showed markedly increase with a dose-dependent manner. Correspondingly, the microscopically pathological differences of the liver tissue observed by HE and oil red O staining indicated that the liver cell swelling, hydropic degeneration and lipid droplets formation induced by alcohol were significantly improved, which suggested the Liuwei Wuling tablet can reduce the liver injure. In conclusion, the Liuwei Wuling tablet had the protective effect on acute alcoholic hepatic injury which maybe depended on the mechanism of relieving lipid peroxidation, elevating antioxidant enzymes activity, inhibiting oxidative stress and reducing inflammation factors expression.