Lactiplantibacillus plantarum LP4 alleviates alcoholic liver disease in C57BL/6 mice.

Cell Type–Dependent Pro- and Anti-Inflammatory Role of Signal Transducer and Activator of Transcription 3 in Alcoholic Liver Injury

Alcoholic liver disease: a matter of hormones?

Traditional treatments have a poor effect on alcoholic liver diseases. Linderae radix (LR), the dried root of Lindera aggregata (Sims) Kosterm., has been frequently used in traditional Chinese medicine for treating various diseases, and has been shown to exhibit a protective effect on liver injury. In the present study, LR extracts were made using various solvents, and then administrated to rats to establish a model of ethanol-induced liver injury. The study aimed to investigate the therapeutic effects and potential mechanism of LR extracts on acute alcoholic liver injury. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), triglycercide (TG), cholesterol (TC), methane dicarboxylic aldehyde (MDA) and superoxide dismutase (SOD) were determined using an automatic biochemistry analyzer. In addition, pathological examination was performed by hematoxylin-eosin staining. The levels of MDA and SOD, and the expression levels of nuclear factor (NF)-κB, tumor necrosis factor (TNF)-α and interleukin (IL)-1β in liver tissue were investigated immunohistochemically. The expression of cytochrome P450 2E1 (CYP2E1) mRNA was quantified by reverse transcription-quantitative polymerase chain reaction. The results indicated that LR extracts improved the histopathological status and decreased the serum levels of ALT, AST, TG, TC and MDA. Furthermore, the levels of MDA and inflammatory mediators (NF-κB, TNF-α and IL-1β) were decreased in liver tissues, and the overexpression of CYP2E1 mRNA induced by ethanol treatment. LR extracts exhibited a protective effect on alcoholic liver injury and the mechanism may be associated with the anti-inflammatory and anti-oxidative action.

Background:Anoectochilus roxburghii (Orchidaceae) is a traditional Chinese medicinal herb with anti-inflammatory, antilipemic, liver protective, immunomodulatory, and other pharmacological activities. Kinsenoside (KD), which shows protective effects against a variety types of liver damage, is an active ingredient extracted from A. roxburghii. However, the liver protective effects and potential mechanisms of KD in alcoholic liver disease (ALD) remain unclear. This study aimed to investigate the liver protective activity and potential mechanisms of KD in ALD.Methods: AML12 normal mouse hepatocyte cells were used to detect the protective effect of KD against ethanol-induced cell damage. An alcoholic liver injury model was induced by feeding male C57BL/6J mice with an ethanol-containing liquid diet, in combination with intraperitoneal administration of 5% carbon tetrachloride (CCl4) in olive oil. Mice were divided into control, model, silymarin (positive control), and two KD groups, treated with different doses. After treatment, hematoxylin–eosin and Masson’s trichrome staining of liver tissues was performed, and serum alanine aminotransferase (ALT) and aspartate transaminase (AST) levels were determined to assess the protective effect of KD against alcoholic liver injury. Moreover, proteomics techniques were used to explore the potential mechanism of KD action, and ELISA assay, immunohistochemistry, TUNEL assay, and western blotting were used to verify the mechanism.Results: The results showed that KD concentration-dependently reduced ethanol-induced lipid accumulation in AML12 cells. In ALD mice model, the histological examination of liver tissues, combined with the determination of ALT and AST serum levels, demonstrated a protective effect of KD in the alcoholic liver injury mice. In addition, KD treatment markedly enhanced the antioxidant capacity and reduced the endoplasmic reticulum (ER) stress, inflammation, and apoptosis compared with those in the model group. Furthermore, KD increased the phosphorylation level of AMP-activated protein kinase (AMPK), inhibited the mechanistic target of rapamycin, promoted the phosphorylation of ULK1 (Ser555), increased the level of the autophagy marker LC3A/B, and restored ethanol-suppressed autophagic flux, thus activating AMPK-dependent autophagy.Conclusion: This study indicates that KD alleviates alcoholic liver injury by reducing oxidative stress and ER stress, while activating AMPK-dependent autophagy. All results suggested that KD may be a potential therapeutic agent for ALD.

Zinc Supplementation Prevents Alcoholic Liver Injury in Mice through Attenuation of Oxidative Stress

Alcohol consumption alters the diversity and metabolic activities of gut microbiota, leading to intestinal barrier dysfunction and contributing to the development of alcoholic liver disease (ALD), which is the most prevalent cause of advanced liver diseases. In this study, we investigated the protective effects and action mechanism of an aqueous extraction of Pericarpium citri reticulatae and Amomi fructus (PFE) on alcoholic liver injury. C57BL/6 mice were used to establish the mouse model of alcoholic liver injury and orally administered 500 and 1,000mg/kg/d of PFE for 2weeks. Histopathology, immunohistochemistry, immunofluorescence, Western blotting, qRT-PCR, and 16S rDNA amplicon sequencing were used to analyze the mechanism of action of PFE in the treatment of alcohol-induced liver injury. Treatment with PFE significantly improved alcohol-induced liver injury, as illustrated by the normalization of serum alanine aminotransferase, aspartate aminotransferase, total triglyceride, and cholesterol levels in ALD mice in a dose-dependent manner. Administration of PFE not only maintained the intestinal barrier integrity prominently by upregulating mucous production and tight junction protein expressions but also sensibly reversed the dysregulation of intestinal microecology in alcohol-treated mice. Furthermore, PFE treatment significantly reduced hepatic lipopolysaccharide (LPS) and attenuated oxidative stress as well as inflammation related to the TLR4/NF-κB signaling pathway. The PFE supplementation also significantly promoted the production of short-chain fatty acids (SCFAs) in the ALD mice. Administration of PFE effectively prevents alcohol-induced liver injury and may also regulate the LPS-involved gut-liver axis; this could provide valuable insights for the development of drugs to prevent and treat ALD.

Chronic alcohol consumption is a leading cause of chronic liver disease worldwide, leading to cirrhosis and hepatocellular carcinoma. Currently, the most widely used model for alcoholic liver injury is ad libitum feeding with the Lieber-DeCarli liquid diet containing ethanol for 4-6 weeks; however, this model, without the addition of a secondary insult, only induces mild steatosis, slight elevation of serum alanine transaminase (ALT) and little or no inflammation. Here we describe a simple mouse model of alcoholic liver injury by chronic ethanol feeding (10-d ad libitum oral feeding with the Lieber-DeCarli ethanol liquid diet) plus a single binge ethanol feeding. This protocol for chronic-plus-single-binge ethanol feeding synergistically induces liver injury, inflammation and fatty liver, which mimics acute-on-chronic alcoholic liver injury in patients. This feeding protocol can also be extended to chronic feeding for longer periods of time up to 8 weeks plus single or multiple binges. Chronic-binge ethanol feeding leads to high blood alcohol levels; thus, this simple model will be very useful for the study of alcoholic liver disease (ALD) and of other organs damaged by alcohol consumption.

Alcoholic liver disease (ALD), caused by consumption of alcohol, with high morbidity and mortality, whose effective interventions is essential. Chinese medicine has a long history of detoxification, and Chaige anti-alcoholic granules (CAG) is an accepted formula including 13 Chinese herbs with definite detoxifying and liver-protecting effects in clinical for acute alcohol intoxication. However, the underlying mechanism is unclear. In this study, mice were selected for acute toxicity experiments with the increasing drug concentration to 0.78 mg⋅mL-1. Body weight changes, organ indices, liver and kidney histological observations were performed after 2 weeks. ALT, AST, BUN, and creatinine in mice serum were detected by the kits. A rat model of alcoholic liver injury (ALI) was established by gavage of Chinese wine with 56% alcohol, which was intragastrically received with CAG at 1575, 3150 and 6300 mg⋅kg⋅day-1 for 2 weeks, respectively, while positive group 100 mg⋅kg⋅day-1 metadoxine. The organ indices were measured, and the protective effect of CAG on ALI was determined using kits, ELISA, histopathology, and western blotting. The results of the acute toxicity experiment showed that the mice were alive normally and the organ index, liver and kidney histopathology, and serum biochemical indicators showed no significant difference between the control group and the CAG-treated groups. The results of hepatoprotective effect of CAG in rat showed that compared with the control group, the liver index, ALT, AST, ADH, TC, TG, GSH-Px, SOD, and CYP450 2E1 levels were all increased in the model group (P < 0.01), while ALDH and MDA were decreased (P < 0.01). Compared with the model group, the detection indicators in the different dose CAG treatment groups could be reversed, and there was a certain dose-effect relationship, that is, the reversal effect of the high and med-dose groups was better (P < 0.01). Liver histological observation showed that CAG could alleviate the infiltration of inflammatory cells. These indicated that CAG had no acute toxicity and exhibited a large safety range, and was first identified to protect against hepatotoxicity through anti-oxidative stress and anti-inflammation, providing a scientific basis for further research into its clinical applications.

Alcoholic Liver Disease: Pathogenesis and New Therapeutic Targets

Objective To evaluate the hepatoprotective mechanism of Xwak granule (Xwak) in treatment of mice with alcoholic liver injury via activating ERK/NF-κB and Nrf/HO-1 signaling pathways. Methods The chemical composition of Xwak was tested by liquid chromatography coupled with mass spectrometry (LC-MS). Herein, 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging assay and 2,2-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid (ABTS) radical tests were performed in vitro. The hepatoprotective effect of Xwak was assessed at different concentrations (1.5, 3, and 6 g/kg) in a mouse model of alcoholic liver injury. Results Totally, 48 compounds, including 16 flavonoids, 8 tannins, 9 chlorogenic acids, and 15 other compounds, were identified from Xwak. Xwak showed to have a satisfactory antioxidant activity in vitro. In a group of Xwak-treated mice, the serum levels of alanine transaminase (ALT), aspartate transaminase (AST), and alkaline phosphatase (ALP) were decreased compared with a group of the mouse model of alcoholic liver injury. In addition, the levels of antioxidant enzymes, such as glutathione peroxidase (GSH-PX), total superoxide dismutase (T-SOD), and catalase (CAT), were noticeably increased and the levels of malondialdehyde (MDA), tumor necrosis factor-α (TNF-α), transforming growth factor-β (TGF-β), and interleukin-6 (IL-6) were markedly reduced in the liver of mice. The state of oxidative stress in the mouse model of alcoholic liver injury was improved after treatment with Xwak. The improvement of inflammation-mediated disruption may conducive to the Xwak activity in the control of liver injury. The signals of p-ERK1/2, p-NF-κB, COX-2, iNOS, CYP2E1, Nrf, and HO-1 were significantly induced in the liver of mice after treatment with Xwak. Conclusions The abovementioned findings indicated that the hepatoprotective mechanism of Xwak could be achieved by activating ERK/NF-κB and Nrf/HO-1 signaling pathways to alleviate oxidative stress and inflammatory.

The occurrence of alcoholic liver injury is related to the oxidative stress. Bacteria for alleviating alcoholic related liver injury have received widespread attention. Study aims to investigate the alleviated efficacy of Lactiplantibacillus plantarum (L. plantarum) P101 on alcohol-induced liver injury and its potential mechanism. The model of alcoholic liver injury was obtained according to the NIAAA method and the mice were treated with L. plantarum P101 (108 CFU.mice-1). Results showed that treatment of L. plantarum P101 could significantly improve liver function and antioxidant capacity. Furthermore, L. plantarum P101 significantly up-regulated Nuclear factor erythroid 2-related factor (Nrf2) and its target molecule, Hemeoxygenase 1 (HO-1), by promoting nuclear translocation of Nrf2. Moreover, inflammatory factors and pro-apoptotic protein (Caspase3) levels were significantly decreased in mice treated with L. plantarum P101. This study confirmed that the beneficial effect of L. plantarum P101 supplement was achieved via regulating Nrf2/HO-1 antioxidant pathway, and alleviated alcoholic liver injury.

Alcoholic liver disease (ALD) remains a major global health challenge, necessitating effective therapeutic interventions. Polyene phosphatidyl choline (PPC) has shown potential in liver disorders, yet its clinical efficacy and molecular mechanisms in ALD require further validation. This study aimed to assess the clinical effectiveness of PPC using real-world data and to elucidate its hepatoprotective mechanisms in a murine ALD model. Clinical efficacy was evaluated via retrospective data analysis, focusing on PPC's effects on liver injury and lipid metabolism biomarkers. A mouse model of alcoholic liver injury was established, and PPC's protective effects were investigated using biochemical assays and histopathology. RNA-sequencing identified differentially expressed genes and pathways, with key targets validated by Western blot. Clinical analysis showed PPC significantly improved liver function and lipid parameters in ALD patients. In mice, PPC alleviated alcohol-induced hepatic inflammation, oxidative stress, and lipid accumulation. RNA-seq indicated involvement of the Akt-PDE3-PKA axis, and Western blot confirmed PPC downregulated Akt1, Traf3, Prkaca, and NF-κB p65 while upregulating Pde3b. Histopathology consistently revealed reduced steatosis and liver damage. In conclusion, PPC injections exert clinically meaningful hepatoprotective effects, likely through regulation of the Akt-PDE3-PKA pathway to modulate lipid homeostasis in ALD.

A polysaccharide from Alhagi honey protects the intestinal barrier and regulates the Nrf2/HO-1-TLR4/MAPK signaling pathway to treat alcoholic liver disease in mice

Alcoholic liver damage–toxicity, autoimmunity and allergy

Hyperactivation of mammalian target of rapamycin complex 1 (mTORC1) and endoplasmic reticulum (ER) stress promote hepatic lipid accumulation and insulin resistance. To elucidate the role of mTORC1 in the alcoholic liver disease (ALD), we examined mTORC1 activation and adaptive unfolded protein response (UPR) mechanisms in chronic plus binge ethanol feeding (CB) murine model of alcoholic liver injury. CB resulted in hepatic steatosis, elevation of serum alanine aminotransferase levels, exacerbation of cell apoptosis as indicated as caspase 3 cleavage and poly ADP-ribose polymerase activation. In livers of CB mice, selective UPR genes were upregulated, as evidenced by X-box binding protein splicing, phosphorylation of eIF2α and increased levels of ER chaperone. Furthermore, CB led to defective adaption to prolonged ER stress for ER stress-mediated apoptosis was activated in liver. Similar ER stress responses were achieved/attenuated in mice after challenging with tunicamycin/4-PBA, an ER stress inducer/inhibitor. The steatotic changes were also attributed to lipids accumulation as reflected by sterol regulatory element-binding protein 1 and peroxisome proliferator-activated receptor alpha. Importantly, the activity of mTORC1, as indicated by phosphorylation of S6K1 and 4E-BP1, was increased in CB mice. mTORC1 inhibition by Deptor overexpression or rapamycin injection can restore hepatic ER homeostasis, inhibited lipogenesis, and reduced fatty liver. Specific signaling pathway regulated by mTORC1 was also determined in S6K1 dominant negative CB mice. Therefore, hepatic activation of mTORC1 may play a causal role in ER stress and steatosis related to ALD.