Alcoholic Injury Research Articles

Protective Effects of Polysaccharides From Sargassum hemiphyllum (Turner) C. Agardh Against Alcohol‐Induced LO2 Cell Damage

ABSTRACTThe study aimed to explore the protective impact of polysaccharide derived from Sargassum hemiphyllum (Turner) C. Agardh (SHP) against ethanol‐induced injury in LO2 hepatocytes, along with its potential mechanism of action. A model of alcoholic injury in LO2 cells was established to assess the shielding effect of SHP against liver injury induced by alcohol. Treatment with 800 mmol/L ethanol for 6 h was selected for the construction of the hepatocyte injury model. Compared with those in the alcohol model group, the survival rate of LO2 cells in the SHP treatment group was significantly greater. When the concentration of SHP reached 60 μg/mL, the cell viability increased to 89.17% ± 3.58%. Moreover, SHP treatment significantly reduced the level of intracellular reactive oxygen species (ROS), increased the levels of intracellular glutathione (GSH), lactate dehydrogenase (LDH), and catalase (CAT), reduced the level of malondialdehyde (MDA), and prevented the leakage of intrahepatic enzymes (aspartate aminotransferase (AST) and alanine transaminase (ALT)) to protect LO2 cells from alcohol‐induced injury. Moreover, at a concentration of 60 μg/mL, SHP inhibited the ethanol‐induced reduction in the protein expressions of Nrf2, HO‐2, and GCLC, indicating its potential to modulate the antioxidant system to restore the homeostatic state, consequently shielding the liver from peroxidative damage induced by alcohol. These results propose that SHP exhibits a protective role against oxidative damage in LO2 cells and holds promise as a novel natural hepatoprotective agent for averting liver injury.

Gastroprotective potential of the aqueous extract of nine-steaming and nine-sun-drying processed Polygonatum cyrtonema Hua against alcoholic gastric injury in mice

Ethnopharmacological relevancePolygonatum (Huangjing) genus has been used as both food and medicine in China for 2000 years, which was regarded as a “Top-grade” herb in the Shennong Bencao Jing. The most commonly used species is the rhizome of Polygonatum cyrtonema Hua (PC) that is traditionally utilized to invigorate Qi, nourish Yin, moisten lung, and tonify spleen and kidney. Aim of the studyExcessive alcohol consumption causes severe upper-gastrointestinal diseases, notably gastric mucosal damage characterized by hemorrhagic gastritis, which lacks safe and effective intervention. This study aims to investigate the gastroprotective effects of nine-steaming and nine-drying processed Polygonatum cyrtonema Hua (PPC) on alcohol-induced gastric mucosal damage in mice. Materials and methodsPPC extract was chemically characterized by UPLC-QE-MS analysis. ICR mice were subjected to an ethanol-induced gastric lesion model and were orally administered PPC aqueous extract for 5 consecutive days. After treatment, gastric tissues were stained with hematoxylin and eosin (H&E), and the pro-inflammatory and oxidative stress factors were determined using ELISA and Multiplex assay, while the gene expressions of gastric tissues were detected by RNA-seq and Western blotting. ResultsPPC reduced the alcohol concentration of liquor in vitro and protected against alcohol-induced gastric mucosal lesion in mice. Notably, PPC aqueous extract relieved alcohol-induced pro-inflammatory and oxidative stress factors, including interleukin 6 (IL-6), IL-8, keratinocyte-derived chemokine (KC), monocyte chemotactic protein-1 (MCP-1), superoxide dismutase (SOD), glutathione (GSH), and malondialdehyde (MDA). RNA-sequencing analysis revealed that ethanol exposure activated mitogen-activated protein kinases (MAPKs), tumor necrosis factor (TNF), and IL-17 signaling pathways in gastric tissue, and these activated signaling pathways were inhibited by the PPC treatment. Consistently, Western blot data showed that PPC treatment suppressed the activation of extracellular signal-regulated kinases (ERK), p38, c-Jun N-terminal kinases (JNK), TNF-α and IL-17A pathways in gastric tissue. ConclusionIn conclusion, the aqueous extract of PPC exerted a gastroprotective effect against alcohol-induced gastric injury by alleviating inflammation and oxidative stress, potentially through the inhibition of the MAPKs, IL-17 and TNF-α pathways. These findings supported the future development of PPC as an effective intervention for alcohol-induced gastric damage.

The utility of alcohol saliva test strips compared to the breathalyzer in trauma patients in a resource-limited setting.

The correlation between alcohol consumption and injury is undeniable. However, past research relying on self-reporting alcohol use likely resulted in underreporting and emphasizing the need to increase alcohol testing, especially in resource-limited settings where the burden of injuries is highest. This is a prospective analysis of injured patients presenting to the trauma center at Kamuzu Central Hospital in Lilongwe, Malawi. We collected information including patient age, sex, admission date, mechanism of injury, breathalyzer test and Rapid ResponseTM Alcohol Saliva Test Strips (AST) result, and survival. A total of 805 trauma patients were included. The overall prevalence of alcohol consumption in this trauma cohort is 18.3%. There was a 95.5% agreement between the AST and breathalyzer test with a Kappa coefficient of 0.83. The sensitivity and specificity of the AST were determined to be 78.5% (CI 75.7-81.2) and 99.3% (CI 98.7-99.9), respectively. ROC analyses showed the AST to have excellent discrimination with an area under the curve of 0.88 (95% CI 0.85-0.92). The prevalence of alcohol-related injury is high in Malawi and the use of the Alcohol Saliva Test Strips is feasible and correlated with results derived from the breathalyzer. Routine alcohol testing for trauma patients presenting to a resource-limited setting is imperative and should be implemented.

Lactobacillus plantarum LP104 improved intestinal and brain inflammation by modulating TLR4 pathway and gut flora in alcohol liver injury mice

In this study, Lactobacillus plantarum LP104 (LP104) originating from kimchi was treated to C57BL/6N mice fed a chronic-plus-single-binge ethanol diet for ten days, aiming to evaluate the effects of the probiotics on intestinal and brain inflammation in the alcohol-induced liver injury model. Dietary supplementation with LP104 significantly reduced the concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total cholesterol (TC), triglycerides (TG) and low-density lipoprotein cholesterol (LDL-C) in alcoholic hepatic injury mice. The 16S rDNA sequencing results inferred that LP104 modulated the intestinal microbiota by depleting inflammation-related bacterial genera Proteobacteria and enriching Lactobacillus and Roseburia. Furthermore, LP104 intervention attenuated alcohol-induced gut inflammation by the TLR4 signaling pathway, which regulated the expressions of intestinal tight junction proteins such as ZO-1, Claudin-1 and Occludin. Subsequently, LP104 intake exerted neuroprotection by restoring neurotrophic factor levels and the blood-brain barrier (BBB) function to decrease lipopolysaccharide (LPS) levels in the serum. The western blotting results suggested LP104 ameliorated alcohol-induced brain inflammation by inhibiting the LPS/TLR4 signaling pathway in mice. Spearman's correlation analysis demonstrated that differential intestinal bacterial taxa were connected with the expressions of inflammation-related proteins in both gut and brain of alcohol liver injury mice.

Curcumol ameliorates alcohol and high-fat diet-induced fatty liver disease via modulation of the Ceruloplasmin/iron overload/mtDNA signaling pathway.

Fatty liver disease (FLD), a chronic liver disease characterized by excessive lipid deposition, is affecting more and more people worldwide owing to the increasing global incidence of obesity and heavy alcohol consumption. However, there is still no effective strategy for prevention or treatment of alcohol and high-fat diet (HFD)-induced FLD. The purpose of this study was to investigate the effect of curcumol on alcohol and HFD-induced FLD and the underlying molecular mechanisms. The results showed that curcumol ameliorated alcohol and HFD-induced hepatocyte injury in vivo and in vitro, and the mechanism might be related to its up-regulation of ceruloplasmin and subsequent alleviation of iron overload. Moreover, curcumol inhibited alcohol and HFD-induced mitochondrial damage and mtDNA release in hepatocytes by modulating iron overload. Furthermore, curcumol's inhibition of mtDNA release could suppress the activation of cGAS-STING and subsequent inflammation, and this phenomenon could be reversed by cGAS overexpression. Notably, alcohol and HFD-induced mtDNA release from hepatocytes contributed to HSC activation and this effect could be weakened by curcumol. In conclusion, these findings elucidated that curcumol ameliorated alcohol and HFD-induced FLD via modulating ceruloplasmin/iron overload/mtDNA signaling pathway, which lead to the inhibition of inflammation and HSCs activation.

Comparative hepatoprotective effects of flavonoids-rich fractions from flowers and leaves of Penthorum chinense Pursh in vitro

Ethnopharmacological relevancePenthorum chinense Pursh is a traditional Miao ethnomedicine rich in bioactive components, widely recognized for its hepatoprotective properties. However, the hepatoprotective effects of its flowers and leaves have not been individually elucidated. Aims of the studyThe objective of this study was to isolate and purify flavonoids-rich fractions from the flowers (PFF) and leaves (PLF) of P. chinense, and to assess their potential protective effects against oxidative, alcohol-induced, and free fatty acid (FFA) induced injury in hepatic cells. Materials and methodsThe P. chinense flowers and leaves flavonoids-rich fractions were extracted by the method optimized by response surface methodology, and the extracts were subsequently purified using petroleum ether and microporous column. The physical characteristics and component composition of PFF and PLF were analyzed by FT-IR and UPLC-MS/MS. The hepatoprotective activities of PFF and PLF were evaluated by the alcohol, H2O2, and FFA-induced hepatocyte injury cellular model in vitro. The protective effects of PFF and PLF on the hepatic cells were evaluated by assessing cell apoptosis rate, enzymes activities, mitochondrial membrane potential, and mRNA expression in relevant signaling pathways. ResultsThe results revealed that PFF was mainly composed of pinocembrin, quercitrin and quercetin, while PLF was predominantly composed of quercetin, pinocembrin, and kaempferol and their derivatives. PFF and PLF exhibited distinct effects on increasing the cell proliferation rate, regulating the MDA, GOT and GPT levels, and modulating the mRNA expression in apoptosis and antioxidant pathways in alcohol damaged LO2 cells. PFF exhibited superior efficacy in reducing cell apoptosis in alcohol-damage cells compared to PLF. Both PFF and PLF alleviated mitochondrial stress in H2O2-induced LO2 cells. Additionally, the PFF and PLF attenuated lipid accumulation and activated mRNA expressions in PPARα/ACOX1/CPT-1 lipid metabolism pathways, as well as Nrf2/ARE oxidative stress pathways. ConclusionThis study compared the hepatoprotective activities of flavonoids-rich fractions purified from the flowers and leaves of P. chinense. The results contribute to the enhanced development and utilization of various parts of P. chinense aimed at medical and health food applications.

AKR7A5 knockout promote acute liver injury by inducing inflammatory response, oxidative stress and apoptosis in mice.

Alcohol liver disease has become a worldwide critical health problem. The ingested alcohol could be converted into acetaldehyde or combined with free fatty acids to induce the endoplasmic reticulum oxidative stress in the liver. Coincidentally, AKR7A5 has both aldehyde detoxification and antioxidant effects. Therefore, we discuss the possible role and mechanism of AKR7A5 in the acute alcohol injury of mice liver. There were four experiment groups, the C57BL/6 mice of wild-type mice (WT) or AKR7A5-/- mice (KO) were intragastrically administrated with saline or 50% ethanol at 14 mL/kg, respectively. Compared to the WT + alcohol group, abnormal liver function, disordered hepatic cord, severe congestion in the hepatic sinus and the space of the hepatic cord, occurrence of oxidative stress, DNA damage and different expressions of apoptosis-related proteins were detected in the KO + alcohol group. Meanwhile, the biological process enrichment analysis showed that the down-regulated proteins were related to the metabolism of fatty acid, the up-regulated proteins positive regulation of reactive oxygen species metabolic process, negative regulation of coagulation and haemostasis. In conclusion, single ethanol binge combined with the absence of AKR7A5 caused more severe inflammatory response, oxidative stress, apoptosis of endogenous pathways, abnormal lipids metabolism and disordered coagulation in mice liver.

Peucedanum praeruptorum Dunn leaf aqueous extract protects against alcoholic gastric injury by inhibiting inflammation and oxidative stress in mice

Ethnopharmacological relevancePeucedanum praeruptorum Dunn (PPD) was used to treat gastrointestinal disease in China before the Tang Dynasty, and it was considered a “Top-grade” herb in Shennong Bencaojing, known for its ability to relieve the stomach Qi and indigestion. Aim of the studyAlcohol consumption can induce severe gastric mucosal injury that lacks effective and safe interventions. We aimed to investigate the gastroprotective effects of Peucedanum praeruptorum Dunn leaf (PPL) after bolting in alcohol-induced gastric damage in mice. Materials and methodsMice were orally administered PPL aqueous extract at doses of 2.5, 5, and 10 g/kg for 5 consecutive days prior to the induction of gastric damage model with ethanol. Gastric tissue was stained by hematoxylin and eosin (H&E), and the levels of pro-inflammatory cytokines and oxidative stress indicators were determined using ELISA and RT-qPCR. RNA-seq was used to detect differentially expressed genes (DEGs) in the gastric tissue, while Western blotting was employed to measure the expressions of IL-17, TNF-a, and AKT pathways. ResultsTreatment with PPL alleviated alcohol-induced gastric damage in mice, whereas dried root (PPD) and stem (PPS) of Peucedanum praeruptorum Dunn had no gastroprotective function. The content of peucedanocoumarin I was higher in the dried PPL compared to PPD and PPS, with an increase in peucedanocoumarin I content in PPL after boiling. Additionally, PPL administration (5, 10 g/kg) decreased pro-inflammatory factors, such as interleukin-6 (IL-6), IL-8, IL-4, IL-1β, IL-18, and tumor necrosis factor (TNF-a) in alcohol-induced gastric injury mice (p < 0.05), and improved oxidative stress markers, including superoxide dismutase enzymes (SOD), catalase (CAT), and malondialdehyde (MDA) (p < 0.05). RNA-seq data revealed that PPL treatment inhibited alcohol-induced inflammation-related signals, including IL-17 and TNF pathways, and restored alcohol-inhibited gastric digestive and metabolic functions, such as xenobiotics metabolism of cytochrome P450, and protein digestion and absorption pathways. Notably, treatment with PPL downregulated the expressions of IL-17 A, TNF-a, monocyte chemoattractant protein-1 (MCP-1), and AKT-phosphorylation induced by ethanol exposure (p < 0.05). Thus, the aqueous extract of PPL provided protection against alcohol-induced gastric injury by mitigating inflammation and oxidative stress in mice, suggesting a potential novel therapeutic approach for alcohol-induced gastric damage.

Gastroprotective Effect of Isoferulic Acid Derived from Foxtail Millet Bran against Ethanol-Induced Gastric Mucosal Injury by Enhancing GALNT2 Enzyme Activity.

Excessive alcohol consumption has led to the prevalence of gastrointestinal ailments. Alleviating gastric disorders attributed to alcohol-induced thinning of the mucus layer has centered on enhancing mucin secretion as a pivotal approach. In this study, foxtail millet bran polyphenol BPIS was divided into two components with MW < 200 D and MW > 200 D by molecular interception technology. Combined with MTT, cell morphology observation, and trypan blue staining, isoferulic acid (IFA) within the MW < 200 D fraction was determined as the effective constituent to mitigate ethanol-induced damage of gastric epithelial cells. Furthermore, a Wistar rat model with similar clinical features to alcohol-induced gastric mucosal injury was established. Then, gastric morphological observation, H&E staining, and assessments of changes in gastric hexosamine content and gastric wall binding mucus levels were carried out, and the results revealed that IFA (10 mg/Kg) significantly ameliorated alcohol-induced gastric mucosal damage. Finally, we applied techniques including Co-IP, molecular docking, and fluorescence spectroscopy and found that IFA inhibited the alcohol-induced downregulation of N-acetylgalactosamintransferase 2 (GALNT2) activity related to mucus synthesis through direct interaction with GALNT2 in gastric epithelial cells, thus promoting mucin synthesis. Our study lays a foundation for whole grain dietary intervention tailored to individuals suffering from alcoholic gastric mucosal injury.

Mitigative Effect of Graphene Oxide Nanoparticles in Maintaining Gut–Liver Homeostasis against Alcohol Injury

Alcoholic liver disease (ALD) develops when the immunotolerant environment of the liver is compromised due to excessive alcohol consumption. ALD progression involves variations in the expressions of multiple genes, resulting in liver inflammation and the development of a leaky gut. It is still unclear which molecular mechanism is involved in ALD progression, and due to that, there are currently no FDA-approved drugs available for its treatment. In this study, the protective effects of graphene oxide (GO) nanoparticles were investigated against ethanol-induced damage in the gut–liver axis in in vitro. GO was synthesized using a modified Hummer’s method, and characterization was performed. Given the general concerns regarding nanoparticle toxicity, assessments of cell viability, lipid accumulation, DNA damage, cell death, and the generation of reactive oxygen species (ROS) were conducted using various techniques. Furthermore, the gene expressions of pro- and anti-inflammatory cytokines were determined using RT-qPCR. The findings reveal that GO promoted cell viability even against ethanol treatment. Additionally, lipid accumulation significantly decreased when cells were treated with GO alongside ethanol compared to ethanol treatment alone, with similar trends observed for other assays. A gene expression analysis indicated that GO treatment reduced the expression of proinflammatory cytokines while enhancing the expression of antioxidant genes. Moreover, GO treatment led to improvements in gut integrity and a reduction in proinflammatory cytokines in colon cells damaged by ethanol. These findings suggest that GO holds promise as a drug carrier, exhibiting no observed toxic effects. By shedding light on the protective effects of GO against ethanol-induced damage, this study contributes to the burgeoning field of nanoparticle-mediated therapy for ALD.

Potential mechanism of sea cucumber gut peptides in protecting ethanol-induced gastric mucosal injury

Alcoholic gastric injury is a prevalent condition that can result in gastric ulcers. This research delves into the protective effects of sea cucumber gut peptides, Val-Thr-Pro-Tyr (VTPY) and Val-Leu-Leu-Tyr (VLLY), on ethanol-induced gastric injury. These peptides significantly alleviate the pathological symptoms of gastric tissue affected by ethanol, reducing ulcer area and index, epithelial cell loss, submucosal edema, and inflammatory infiltration. High doses of VTPY and VLLY resulted in a significant decrease in ulcer index, with reductions from 5.38 to 2.63 and 1.13, respectively (P < 0.05). Moreover, the levels of P–NF-κB/p65 expression decreased by 33.47% and 54.27%, while myeloperoxidase (MPO) activity decreased by 42.98% and 55.33%, respectively. These treatments also effectively suppressed inflammatory cytokine expression and inhibited macrophage infiltration, ultimately alleviating the inflammatory response in gastric mucosal tissue. They also enhance mitochondrial function by regulating factors associated with mitochondrial dynamics and increasing succinate dehydrogenase (SDH) activity. Furthermore, they alleviate endoplasmic reticulum stress by reducing the immunoglobulin heavy chain-binding protein (Bip), phosphorylated eukaryotic translation initiation factor 2 alpha (P-eIF2α), and activating transcription factor 4 (ATF4) expression levels. Ultimately, high doses of VTPY and VLLY increased the B-cell lymphoma-extra large to B-cell lymphoma-2 associated death promoter ratio (Bcl-XL/Bad ratio) by 138.68% and 247.99%, respectively, while the expression level of cleaved-caspase 3 decreased by 35.26% and 48.22%, thereby preventing the activation of the apoptotic pathway. Sea cucumber gut peptides VTPY and VLLY exhibit effective anti-inflammatory and anti-apoptotic properties, along with anti-endoplasmic reticulum stress effects, by inhibiting the ATF4/Bcl-2 signaling pathway, thus effectively shielding against ethanol-induced gastric damage. VLLY appears to be more potent in this aspect.

Polysaccharides from Eucommia ulmoides Oliv. leaves alleviates alcohol-induced mouse brain injury and BV-2 microglial dysfunction

Acute alcohol intoxication is a harmful clinical condition characterized by behavioral and neurological symptoms, for which few effective therapies are available at present. Dysfunction of microglial BV-2 cells has been reported to be associated with acute alcohol-induced brain injuries. In the present study, the protective effects of Eucommia ulmoides Oliv. leaves polysaccharides (EULP) on acute alcoholic brain injury and microglial dysfunction were investigated. 14-day pretreatment of EULP significantly attenuated neurobehavioral deficit and neurotransmitter damage in the brain tissue of mice caused by acute alcohol exposure. Additionally, EULP regulated the metabolic disorder of brain tissue. Consistently, it was shown that EULP pretreatment significantly improved alcohol-induced phagocytosis decrease, oxidative stress and inflammation in BV-2 cells. Therefore, EULP may be proposed and employed as a potential therapeutic agent for alcohol-induced brain damage.

Preparation of Pueraria lobata Root-Derived Exosome-Like Nanovesicles and Evaluation of Their Effects on Mitigating Alcoholic Intoxication and Promoting Alcohol Metabolism in Mice.

Pueraria lobata (P. lobata), a dual-purpose food and medicine, displays limited efficacy in alcohol detoxification and liver protection, with previous research primarily focused on puerarin in its dried roots. In this study, we investigated the potential effects and mechanisms of fresh P. lobata root-derived exosome-like nanovesicles (P-ELNs) for mitigating alcoholic intoxication, promoting alcohol metabolism effects and protecting the liver in C57BL/6J mice. We isolated P-ELNs from fresh P. lobata root using differential centrifugation and characterized them via transmission electron microscopy, nanoscale particle sizing, ζ potential analysis, and biochemical assays. In Acute Alcoholism (AAI) mice pre-treated with P-ELNs, we evaluated their effects on the timing and duration of the loss of the righting reflex (LORR), liver alcohol metabolism enzymes activity, liver and serum alcohol content, and ferroptosis-related markers. P-ELNs, enriched in proteins, lipids, and small RNAs, exhibited an ideal size (150.7 ± 82.8 nm) and negative surface charge (-31 mV). Pre-treatment with 10 mg/(kg.bw) P-ELNs in both male and female mice significantly prolonged ebriety time, shortened sobriety time, enhanced acetaldehyde dehydrogenase (ALDH) activity while concurrently inhibited alcohol dehydrogenase (ADH) activity, and reduced alcohol content in the liver and serum. Notably, P-ELNs demonstrated more efficacy compared to P-ELNs supernatant fluid (abundant puerarin content), suggesting alternative active components beyond puerarin. Additionally, P-ELNs prevented ferroptosis by inhibiting the reduction of glutathione peroxidase 4 (GPX4) and reduced glutathione (GSH), and suppressing acyl-CoA synthetase long-chain family member 4 (ACSL4) elevation, thereby mitigating pathological liver lipid accumulation. P-ELNs exhibit distinct exosomal characteristics and effectively alleviate alcoholic intoxication, improve alcohol metabolism, suppress ferroptosis, and protect the liver from alcoholic injury. Consequently, P-ELNs hold promise as a therapeutic agent for detoxification, sobriety promotion, and prevention of alcoholic liver injury.

C-Jun N-Terminal Kinase-2 (JNK2) Plays a Crucial Role in Alcohol and Corticosterone-Induced Dysbiosis of Gut Microbiota and Tissue Injury at the Gut-Liver Axis

Introduction: About 15% of heavy drinkers develop liver disease, suggesting the “Multiple hit hypothesis” of Alcohol-Associated Liver Diseases (AALD). Chronic stress is a potential second hit in AALD pathogenesis. Our recent study showed that alcohol-induced gut and liver injury is exacerbated by chronic restraint stress or corticosterone. JNK2 plays an essential role in stress-induced cell injury. In this study, we investigated the role of JNK2 in ethanol (EtOH) and corticosterone-induced gut and liver injury and microbiota dysbiosis in mice. Methods: Wildtype (WT) and JNK2 knockout (JNK2-KO) mice (male and female; 8-10 wks) were fed a Lieber-DiCarli liquid diet with or without EtOH (0% 2d, 1% 2d, 2% 2d, 4% 1wk, 5% 1wk, &amp; 6% 1wk). Control groups were pair-fed isocaloric EtOH-free diet. One group of pair-fed and EtOH-fed WT and JNK2-KO mice were administered corticosterone (CORT, 25 mg/kg/day; s.c.). Intestinal permeability was measured by vascular-to-luminal flux of FITC-inulin in vivo, tight junction (TJ) and adherens junction (AJ) integrity was assessed by confocal microscopy for occludin, ZO-1, E-cadherin, and β-catenin, and mucosal inflammation by RT-PCR for cytokines/chemokines. Plasma lipopolysaccharide (LPS) and cytokines were measured to evaluate endotoxemia and systemic inflammation. Liver damage was assessed by measuring plasma AST/ALT, liver triglyceride, and cytokine/chemokine mRNA. Gut microbiota was analyzed by 16S rRNA sequencing of colonic flushing followed by bioinformatics analyses. Results and conclusion: In WT mice, EtOH feeding increased intestinal permeability, disrupted epithelial TJ and AJ, and elevated mucosal cytokine expression, indicating epithelial barrier dysfunction and mucosal inflammation. EtOH increased plasma LPS and cytokine/chemokine levels, indicating endotoxemia and systemic inflammation. Plasma AST/ALT and liver triglyceride levels were increased, indicating EtOH-induced liver damage. CORT significantly elevated EtOH-induced intestinal permeability, TJ/AJ disruption, endotoxemia, systemic inflammation, and liver damage in WT mice. These effects of EtOH and CORT were absent in JNK2-KO mice. Microbiota α-diversity was higher in JNK2-KO mice than in WT mice, and β-diversity analysis showed that the microbiota composition in WT mice differed from that of KO mice. Microbiota α- and β-diversities were altered by EtOH and CORT in WT mice but not in JNK2-KO mice. The relative abundance of Nocardiaceae and Verrucomicrobiaceae was high, and Ruminococcaceae and Bacillaceae were low in KO mice. CORT decreased the abundance of Caulobacteriaceae and Ruminococcaceae and increased Verrucomicrobiaceae and Lactobacillaceae in WT but not KO mice. EtOH increased Ruminococcaceae and Bacillaceae and decreased Lactobacillaceae in WT mice but not KO mice. CORT-induced modulation of EtOH-induced microbiota dysbiosis was observed in WT mice but not KO mice. In conclusion, these data demonstrate that JNK2 regulates gut microbiota in mice, and JNK2 deficiency confers resistance to alcohol and corticosterone-induced tissue injury and microbiota dysbiosis. Acknowledgements: NIH/NIAAA - R01AA012307, AA029270; VA - IO1BX003014. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

41 miR-146a Mimic Administration Attenuates Lung Inflammation After Alcohol and Burn Injury

41 miR-146a Mimic Administration Attenuates Lung Inflammation After Alcohol and Burn Injury

Oxysophoridine inhibits oxidative stress and inflammation in hepatic fibrosis via regulating Nrf2 and NF-κB pathways

BackgroundHepatic fibrosis (HF) runs through multiple stages of liver diseases and promotes these diseases progression. Oxysophoridine (OSR), derived from Sophora alopecuroides l., is a bioactive alkaloid that has been reported to antagonize alcoholic hepatic injury. However, whether OSR suppresses HF and the mechanisms involved in Nrf2 remain unknown. PurposeSince the dysregulation of inflammation and oxidative stress is responsible for the excessive accumulation of extracellular matrix (ECM) and fibrosis in the liver. We hypothesized that OSR may attenuate HF by inhibiting inflammation and oxidative stress through activating Nrf2 signaling. MethodsIn this study, we employed LPS-stimulated HSC-T6 cells, RAW264.7 cells, and a CCl4-induced C57BL/6 mouse fibrotic model to evaluate its suppressing inflammation and oxidative stress, as well as fibrosis. ResultsThe result showed that OSR significantly reduced α-SMA and TGF-β1 at a low dose of 10 μM in vitro and at a dose of 50 mg/kg in vivo, which is comparable to Silymarin, the only Chinese herbal active ingredient that has been marketed for anti-liver fibrosis. Moreover, OSR effectively suppressed the expression of iNOS at a dose of 10 μM and COX-2 at a dose of 40 μM, respectively. Furthermore, OSR demonstrated inhibitory effects on the IL-1β, IL-6, and TNF-α in vitro and almost extinguished cytokine storm in vivo. OSR exhibited antioxidative effects by reducing MDA and increasing GSH, thereby protecting the cell membrane against oxidative damage and reducing LDH release. Moreover, OSR effectively upregulated the protein levels of Nrf2, HO-1, and p62, but decreased p-NF-κB p65, p-IκBα, and Keap1. Alternatively, mechanisms involved in Nrf2 were verified by siNrf2 interference, siNrf2 interference revealed that the anti-fibrotic effect of OSR was attributed to its activation of Nrf2. ConclusionThe present study provided an effective candidate for HF involved in both activation of Nrf2 and blockage of NF-κB, which has not been reported in the published work. The present study provides new insights for the identification of novel drug development for HF.

Patterns of concomitant traumatic brain injury and ocular trauma in US service members

BackgroundConcomitant traumatic brain injury (TBI) and ocular trauma (OT) are caused by the same physical mechanisms, which may complicate therapeutic intervention if screening and evaluation of each condition are not...

Reduced Expression of miR-146a Potentiates Intestinal Inflammation following Alcohol and Burn Injury.

MicroRNAs (miRNAs) are small noncoding RNA molecules that negatively regulate gene expression. Within the intestinal epithelium, miRNAs play a critical role in gut homeostasis, and aberrant miRNA expression has been implicated in various disorders associated with intestinal inflammation and barrier disruption. In this study, we sought to profile changes in intestinal epithelial cell miRNA expression after alcohol and burn injury and elucidate their impact on inflammation and barrier integrity. Using a mouse model of acute ethanol intoxication and burn injury, we found that small intestinal epithelial cell expression of miR-146a is significantly decreased 1 d following injury. Using invitro studies, we show that reduced miR-146a promotes intestinal epithelial cell inflammation by promoting p38 MAPK signaling via increased levels of its target TRAF6 (TNFR-associated factor 6). Furthermore, we demonstrate that invivo miR-146a overexpression significantly inhibits intestinal inflammation 1 d following combined injury and potentially supports intestinal barrier homeostasis. Overall, this study highlights the important impact that miRNA expression can have on intestinal homeostasis and the valuable potential of harnessing aberrant miRNA expression as a therapeutic target to control intestinal inflammation.

The antioxidant peptides from walnut protein hydrolysates and their protective activity against alcoholic injury.

In this study, walnut protein was hydrolyzed, separated by ultrafiltration, purified by RP-HPLC, identified by LC-MS/MS, and screened by molecular docking to finally obtain three novel antioxidant peptides HGEPGQQQR (1189.584 Da), VAPFPEVFGK (1089.586 Da) and HNVADPQR (949.473 Da). These three peptides exhibited excellent cellular antioxidant activity (CAA) with EC50 values of 0.0120 mg mL-1, 0.0068 mg mL-1, and 0.0069 mg mL-1, respectively, which were superior to that of the positive control GSH (EC50: 0.0122 mg mL-1). In the ethanol injury model, three antioxidant peptides enhanced the survival of cells treated with ethanol from 47.36% to 62.69%, 57.06% and 71.64%, respectively. Molecular docking results showed that the three antioxidant peptides could effectively bind to Keap1, CYP2E1 and TLR4 proteins. These results suggested that walnut-derived antioxidant peptides could be potential antioxidants and hepatoprotective agents for application in functional foods.

Research progress of traditional Chinese medicine and its monomer compounds and extracts on alcoholic gastric injury

Research progress of traditional Chinese medicine and its monomer compounds and extracts on alcoholic gastric injury