Ethanol Treatment Research Articles

Sex-dependent effects of chronic intermittent ethanol treatment in Wistar rats

Multiple behavioral parameters are associated with vulnerability to alcohol dependence, but the contribution of such factors as cognitive flexibility and sex differences still require further clarification. Our goal was to assess how sex differences mediate relationships between cognitive flexibility, affective state, hierarchy and ethanol preference before and after chronic intermittent ethanol (CIE) exposure in Wistar rats. The results of our study revealed sex-dependent changes in ethanol preference and locomotor activity throughout the time. Anxiety levels were sex-dependently affected by ethanol exposure with a dramatic decrease in ethanol-exposed females. Similarly, striatal tyrosine hydroxylase expression was affected by CIE in a sex-dependent manner, with ethanol-exposed females having higher expression than males. Baseline cognitive flexibility was better in rats with higher locomotor activity and lower anxiety levels but did not correlate with ethanol preference in rats. Taken together, these results are important for understanding sex-dependent alterations in brain and behavior induced by alcohol. This provides important insights into understanding sex differences in the risk for mental disorders.

Contribution of the yeast bi-chaperone system in the restoration of the RNA helicase Ded1 and translational activity under severe ethanol stress

Preexposure to mild stress often improves cellular tolerance to subsequent severe stress. Severe ethanol stress (10% v/v) causes persistent and pronounced translation repression in Saccharomyces cerevisiae. However, it remains unclear whether preexposure to mild stress can mitigate translation repression in yeast cells under severe ethanol stress. We found that the translational activity of yeast cells pretreated with 6% (v/v) ethanol was initially significantly repressed under subsequent 10% ethanol but was then gradually restored even under severe ethanol stress. We also found that 10% ethanol caused the aggregation of Ded1, which plays a key role in translation initiation as a DEAD-box RNA helicase. Pretreatment with 6% ethanol led to the gradual disaggregation of Ded1 under subsequent 10% ethanol treatment in wild-type cells but not in fes1Δhsp104Δ cells, which are deficient in Hsp104 with significantly reduced capacity for Hsp70. Hsp104 and Hsp70 are key components of the bi-chaperone system that play a role in yeast protein quality control. fes1Δhsp104Δ cells did not restore translational activity under 10% ethanol, even after pretreatment with 6% ethanol. These results indicate that the regeneration of Ded1 through the bi-chaperone system leads to the gradual restoration of translational activity under continuous severe stress. This study provides new insights into the acquired tolerance of yeast cells to severe ethanol stress and the resilience of their translational activity.

Effects of Exogenous Ethanol Treatment in Nutrient Solution on Growth and Secondary Metabolite Contents of Three Herb Species in an Indoor Vertical Farming System.

This study aimed to explore the possibility of exogenous ethanol treatment as a technology to regulate the growth and the synthesis of secondary metabolites in herbaceous plants. After transplantation, sweet basil, Korean mint, and sweet wormwood were cultivated in a controlled vertical farming system and consistently exposed to exogenous ethanol at concentrations of 0, 0.5, 1, 2, 4, and 8 mM. Their growth parameters, antioxidant activity, and secondary metabolite contents were Everything is fine. measured to investigate the effects of the exogenous ethanol treatment on the three plants. The low-concentration ethanol treatments increased the shoot dry weight of the sweet basil and sweet wormwood compared to that of the control. As the ethanol concentration increased, the shoot fresh weight and leaf area in the sweet basil and Korean mint decreased compared to those of the control (0 mM). The DPPH (2,2-Diphenyl-1-picrylhydrazyl) radical scavenging activity and total phenolic content of the three plants increased with the ethanol concentration, while the total flavonoid content did not demonstrate a significant trend. The chlorophyll and carotenoids of the basil showed no apparent concentration-dependent trends; however, the chlorophyll and carotenoids of the Korean mint and sweet wormwood decreased with high ethanol concentrations. Moreover, the antioxidant enzyme activity increased with high ethanol concentrations, indicating that high ethanol concentrations induce oxidative stress in plants.

MicroRNA-124a regulates the differentiation of bone marrow mesenchymal stem cells into neurons

Objective This study investigated the effects of microRNA-124a on the differentiation of bone marrow mesenchymal stem cells (BMSCs) and its underlying mechanism. Methods Flow cytometry was used for isolation and identification of BMSCs. Real-time polymerase chain reaction (RT-PCR) was used to detect gene mRNA expression. Apoptosis was detected using Annexin V-FITC/PI Apoptosis Detection Kit. Cell proliferation ability was tested using Cell Counting Kit-8 (CCK-8). The differentiation of BMSCs into neuron inducers β-thiol ethanol or baicalin formed the basis of the study. Results β-thiol ethanol markedly suppressed the microRNA-124a expression of BMSCs, baicalin markedly induced the microRNA-124a expression of BMSCs and β-thiol ethanol or baicalin promoted apoptosis and reduced the growth of BMSCs. Only the microRNA-124a inhibitor did not affect apoptosis or the differentiation of BMSCs, and it increased the effects of β-thiol ethanol or baicalin on the apoptosis of BMSCs. Conclusion β-thiol ethanol and baicalin treatment could affect microRNA-124a expression in BMSCs. We demonstrated that microRNA-124a promoted the differentiation of BMSCs into neurons.

Carbetocin Inhibits Behavioral Sensitization to Ethanol in Male and Female Mice, Independent of Corticosterone Levels.

Oxytocin (OXT), a pro-social peptide, is increasingly recognized as a potential protective substance against drug addiction. In the context of ethanol, previous research has shown OXT's properties in reducing self-administration, alleviating motor impairment in rodents, and reducing craving in humans. However, its role in behavioral sensitization, a neuroadaptive response resulting from repeated drug exposure linked to an increased drug incentive, remains unexplored. OXT is recognized for its role in regulating the hypothalamic-pituitary-adrenal (HPA) axis, in which corticosterone is acknowledged as a significant factor in the development of behavioral sensitization. This study aimed to investigate the effects of carbetocin (CBT), an analogue of OXT, on the expression of behavioral sensitization to ethanol and the concurrent alterations in plasma corticosterone levels in male and female Swiss mice. We also aimed to confirm previous studies on OXT's impact on ethanol consumption in male mice, but with a focus on CBT, using the two-bottle choice model and the drinking in the dark (DID) methodology. For the sensitization study, the mice received either ethanol (1.8 g/kg, i.p.) or saline treatments daily for 15 consecutive days, followed by treatment with carbetocin (0.64 mg/kg, i.p.) or a vehicle for 6 days. Subsequently, on day 22, all the animals underwent an ethanol challenge to assess the expression of behavioral sensitization. The plasma corticosterone levels were measured on days 21 and 22. The CBT effectively prevented the expression of ethanol-induced behavioral sensitization in both male and female subjects, with no alterations having been detected in their corticosterone levels. In the ethanol consumption study, following an initial phase of ethanol acquisition, the male mice underwent a 6-day treatment with CBT i.p. or saline before being re-exposed to ethanol. We also found a reduction in their ethanol consumption due to the CBT treatment. In conclusion, carbetocin emerges as a promising and effective intervention for mitigating ethanol-induced behavioral sensitization and reducing ethanol intake, highlighting its potential significance in alcohol addiction treatment.

Controlling Argas arboreus and A. persicus (Acari: Argasidae) by Adiantum capillus-veneris L. extracts with phytochemical analysis

Argas species are parasites associated mostly with birds. Their infestations of the host may cause blood loss, resulting in anemia and finally death. Egypt loses millions of tons annually from birds because of these parasites. In addition, they can transmit pathogens to animals and humans. The acaricidal effects of the ethanolic and methanolic extracts of Adiantum capillus-veneris at different concentrations (1–4%) against semi-fed adults of Argas arboreus and A. persicus were investigated during 30 days after treatments. Mobility and mortality, acaricide efficacy, and the concentration that kills 50% of specimens (LC50) were estimated. The percentage of dead adults of both Argas species appeared during 6 days considerably until 30 days was significantly increased after treatment of either ethanol or methanol extracts of Adiantum at 1–4%, versus control groups. Ethanolic extracts (100% mortality) were more effective than methanolic ones (90% mortality) for both Argas species. Argas arboreus (80% efficacy and 5.9% LC50) was more resistant than A. persicus (100% efficacy and 4.1% LC50). Generally, males were more resistant than females. The chemical profile (by gas chromatography-mass spectrometry analysis) for the ethanolic extract of Ad. capillus-veneris at 4% (the most effective extract) was provided for the first time, which showed that the major group was sugars and sugar alcohols, and the main components were thymol-ß-d-glucopyranoside, D-(-)-Tagatofuranose, D-Arabinose, D-Galactose, D-(-)-Fructofuranose and Anthracene, 1-methyl. The efficiency of all these components was discussed. Based on the findings, bioactive compounds present in Ad. capillus-veneris have the potential to be applied as substitutes for synthetic acaricides and a biological control agent in the management of A. arboreus and A. persicus ticks.

Effect of ethanol treatment on the structural, techno-functional, and antioxidant properties of edible insect protein obtained from Tenebrio molitor larvae

Edible insect-derived proteins have attracted considerable attention in the food industry owing to their excellent nutritional and bio-functional activities. Herein, ethanol (20, 40, 60, and 80 %)-treated Tenebrio molitor protein (ETMP) was prepared, and its structural, techno-functional, and antioxidant properties were assessed. As the ethanol concentration increased, the molecular weight of the ETMP decreased, and α-helix content decreased whereas that of β-sheet increased, affecting the secondary structure. Ethanol treatment also resulted in changes in the techno-functional properties of edible insect proteins. ETMP showed significant 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical-scavenging activity (p < 0.05), and its antioxidant activity effectively increased the viability of Vero cells damaged by oxidative stress; 20 % ethanol treatment induced the highest antioxidant activity. In conclusion, our results suggest that appropriate ethanol treatment (20 %) increases the antioxidant activity of edible insect proteins, suggesting their potential in food as an alternative protein resource and functional food with excellent antioxidant activity.

Low-dose ethanol consumption inhibits neutrophil extracellular traps formation to alleviate rheumatoid arthritis

Rheumatoid arthritis (RA) is a chronic systemic autoimmune disease. Ethanol consumption has been reported to reduce morbidity in RA patients, but the mechanism behind it remains unclear. Our results showed that Muribaculaceae was predominant in the gut microbiota of mice after ethanol treatment, and the levels of microbiota metabolite acetate were increased. Acetate reduced arthritis severity in collagen-induced arthritis (CIA) mice, which was associated with a decrease in the articular neutrophils and the myeloperoxidase-deoxyribonucleic acid complex in serum. Meanwhile, in vitro experiments confirmed that acetate affected neutrophil activity by acting on G-protein-coupled receptor 43, which reduced endoplasmic reticulum stress in neutrophils and inhibited neutrophil extracellular traps formation. Furthermore, exogenous acetate reversed CIA mice with exacerbated gut microbial disruption, further confirming that the effect of gut microbial metabolite acetate on neutrophils in vivo is crucial for the immune regulation. Our findings illuminate the metabolic and cellular mechanisms of the gut-joint axis in the regulation of autoimmune arthritis, and may offer alternative avenues to replicate or induce the joint-protective benefits of ethanol without associated detrimental effects.

Alcohol‐Treated Nickel‐Aluminium Catalyst for One‐Step Highly‐Selective Butane‐1,4‐Diol Synthesis from 2‐Butyne‐1,4‐Diol

AbstractThe development of low‐cost nickel‐based catalysts for direct and selective hydrogenation of 2‐butyne‐1,4‐diol (BYD) to butane‐1,4‐diol (BAD) under mild conditions is an important and attractive target both in fundamental research and industrialization but remains a formidable challenge. The primary industrial production method for BAD synthesis is a two‐step reaction route, which suffers from complicated catalysis conditions and high equipment costs. Herein, we develop a high‐performance catalyst via a facile alcohol‐treated strategy for highly selective BAD synthesis at moderate operation conditions. The as‐synthesized NA‐80E catalyst exhibits outstanding BAD selectivity of 98.82 % and BYD conversion of 100 % at 60 °C and 4 MPa, outperforming most reported results for BAD formation in a one‐step process and even being comparable to those obtained by the two‐step hydrogenation reaction route under much high temperatures and pressures. Crucially, we found that after facile alcohol (ethanol) treatment, an intriguing phenomenon of suppression of adjacent acid‐assisted hydrogenolysis via extra acidic Al species at the NiO‐Al2O3 interface is observed, contributing to the precise enhancement of BAD selectivity by inhibiting the production of butanol (BOL). This facile alcohol‐treated method along with the revealed mechanism of blocked hydrogenolysis opens vast possibilities for designing high‐performance and highly‐selective hydrogenation catalysts.

Ameliorative effect of diazepam against ethanol-induced mitochondrial disruption in brains of the mice

Brain oxidative damage and neurodegeneration by ethanol (ETH) are considered as important factors that triggered by oxidative stress. Recently, the abuse of diazepam (DZM) has increased by alcoholism-addicted patients. The present study evaluated the effects of combination treatment of ETH with DZM on oxidative damage induced in brain mitochondria of the mice. Only ETH (0.3, 0.6, and 2.5 g / kg) and ETH+ DZM (2.5 mg / kg) were administered intraperitoneally (ip) to the mice. Pathological changes and oxidative stress biomarkers including ROS, lipid peroxidation, carbonyl protein, mitochondrial function, and glutathione content were evaluated in brain mitochondria after 42 days. Results indicated that co-treatment of DZM and ETH significantly reduced mitochondrial toxicity, oxidative damage, pathological changes and increased level of glutathione. Subchronic ETH administration induced brain oxidative damage, mitochondrial disruption, and serious damage to the brain cells. Whereas, combination treatment improved oxidative damage, mitochondrial function, and pathological changes in brain cells after intoxication by ETH. These findings suggest antioxidant effect of DZM in combination with ETH and can be considered in reducing oxidative stress and mitochondrial damage attenuation in the brain. Combination therapy may be a better therapeutic candidate for prevention of brain oxidative damage induced by ETH.

Deletion of NADPH oxidase 2 in chondrocytes exacerbates ethanol-mediated growth plate disruption in mice without major effects on bone architecture or gene expression.

Excess reactive oxygen species generated by NADPH oxidase 2 (Nox2) in response to ethanol exposure mediate aspects of skeletal toxicity including increased osteoclast differentiation and activity. Because perturbation of chondrocyte differentiation in the growth plate by ethanol could be prevented by dietary antioxidants, we hypothesized that Nox2 in the growth plate was involved in ethanol-associated reductions in longitudinal bone growth. Nox2 conditional knockout mice were generated, where the essential catalytic subunit of Nox2, cytochrome B-245 beta chain (Cybb), is deleted in chondrocytes using a Cre-Lox model with Cre expressed from the collagen 2a1 promoter (Col2a1-Cre). Wild-type and Cre-Lox mice were fed an ethanol Lieber-DeCarli-based diet or pair-fed a control diet for 8 weeks. Ethanol treatment significantly reduced the number of proliferating chondrocytes in the growth plate, enhanced bone marrow adiposity, shortened femurs, reduced body length, reduced cortical bone volume, and decreased mRNA levels of a number of osteoblast and chondrocyte genes. Conditional knockout of Nox2 enzymatic activity in chondrocytes did not consistently prevent any ethanol effects. Rather, knockout mice had fewer proliferating chondrocytes than wild-type mice in both the ethanol- and control-fed animals. Additional analysis of tibia samples from Nox4 knockout mice showed that loss of Nox4 activity also reduced the number of proliferating chondrocytes and altered chondrocyte size in the growth plate. Although Nox enzymatic activity regulates growth plate development, ethanol-associated disruption of the growth plate morphology is independent of ethanol-mediated increases in Nox2 activity.

Myrrh Attenuates Gonadal Toxicity and DNA Fragmentation Induced by Ethanol in Male Rats

Background: Long-term ethanol consumption causes pathological alterations to the male reproductive system, possibly resulting in infertility. Myrrh is a resin with antioxidant properties that is produced from the bark of Commiphora tree species. This research aimed to observe the protective impact of myrrh extract supplementation on male reproductive injury and toxicity induced by ethanol treatment. Methods: Adult male Sprague Dawley rats were orally treated with 40% ethanol (3 g/kg) in combination with myrrh extract (500 mg/kg) for four weeks to investigate the curative efficiency of myrrh extract against ethanol toxicity from physiological, histopathological and genetic aspects. Result: The findings revealed that the sperm count, testosterone level and DNA concentration diminished significantly in the ethanol-treated rats compared to control rats, while the contents of malondialdehyde (MDA), heat shock protein 70 (HSP70) and proinflammatory mediators, including tumor necrosis factor-α (TNF-α) and nitric oxide (NO), were elevated significantly; severe degenerative changes in the testicular structure and strong testicular DNA fragmentation were also observed. Co-treatment with myrrh attenuated lipid peroxidation and improved testicular histomorphology, testosterone levels and DNA concentrations and pattern, while at the same time, the levels of both HSP70 and TNF-α levels were increased. Our results provide insight into the curative impacts of myrrh on ethanol toxicity resulting from the rich content of curzerene in myrrh, which has antioxidant activities.

Silk fibroin hydrogel membranes prepared by a sequential cross-linking strategy for guided bone regeneration

Silk fibroin hydrogel is a widely used material for tissue engineering. However, its mechanical properties are the main obstacle to its application to medical bone regeneration barrier membranes. Here, we developed a new hydrogel membrane for guided bone regeneration (GBR). In this study, a sequential crosslinking strategy including photo crosslinking and organic solvent (ethanol) treatment was used to obtain silk fibroin hydrogel membrane (EA-SF). The mechanical properties of EA-SF were significantly enhanced compared to the hydrogel prepared only by photocrosslinking (E-SF). The compressive and tensile strengths of the hydrogel film treated with 75% ethanol for 6 h were 1.18 ± 0.36 MPa and 0.43 ± 0.03 MPa, respectively. In vitro cell culture results showed that EA-SF has good biocompatibility and can effectively shield fibroblasts (L929). EA-SF also has excellent in vitro protein hydrolysis stability. In vivo experiments (subcutaneous implantation and calvarial defect experiment) confirmed the stability and barrier functionality of EA-SF. In conclusion, this study demonstrated that ethanol could improve the mechanical properties of silk fibroin hydrogel and extend the scope of their application, making silk fibroin hydrogel a promising GBR material.

Autoclavable Albumin-Based Cryogels with Uncompromising Properties.

The development of autoclavable hydrogels has been driven by the need for materials that can withstand the rigors of sterilization without compromising their properties or functionality. Many conventional hydrogels cannot withstand autoclave treatment owing to the breakdown of their composition or structure under the high-temperature and high-pressure environment of autoclaving. Here, the effect of autoclaving on the physical, mechanical, and biological properties of bovine serum albumin methacryloyl (BSAMA) cryogels at three protein concentrations (3, 5, and 10%) was extensively studied. We found that BSAMA cryogels at three concentrations remained little changed after autoclaving in terms of gross shape, pore structure, and protein secondary structure. Young's modulus of autoclaved BSAMA cryogels (BSAMAA) at low concentrations (3 and 5%) was similar to that of BSAMA cryogels, whereas 10% BSAMAA exhibited a higher Young's modulus value, compared with 10% BSAMA. Interestingly, BSAMAA cryogels prolonged degradation. Importantly, cell viability, drug release, and hemolytic behaviors were found to be similar among the pre- and post-autoclaved cryogels. Above all, autoclaving proved to be more effective in sterilizing BSAMA cryogels from bacteria contamination than UV and ethanol treatments. Thus, autoclavable BSAMA cryogels with uncompromising properties would be useful for biomedical applications.

Short-term exposure to ethanol induces transcriptional changes in nontumorigenic breast cells.

Breast cancer is a leading cause of cancer-related deaths in women. Many genetic and behavioral risk factors can contribute to the initiation and progression of breast cancer, one being alcohol consumption. Numerous epidemiological studies have established a positive correlation between alcohol consumption and breast cancer; however, the molecular basis for this link remains ill defined. Elucidating ethanol-induced changes to global transcriptional programming in breast cells is important to ultimately understand how alcohol and breast cancer are connected mechanistically. We investigated induced transcriptional changes in response to a short cellular exposure to moderate levels of alcohol. We treated the nontumorigenic breast cell line MCF10A and the tumorigenic breast cell lines MDA-MB-231 and MCF7, with ethanol for 6 h, and then captured the changes to ongoing transcription using 4-thiouridine metabolic labeling followed by deep sequencing. Only the MCF10A cell line exhibited statistically significant changes in newly transcribed RNA in response to ethanol treatment. Further experiments revealed that some ethanol-upregulated genes are sensitive to the dose of alcohol treatment, while others are not. Gene Ontology and biochemical pathway analyses revealed that ethanol-upregulated genes in MCF10A cells are enriched in biological functions that could contribute to cancer development.

Effect of ethanol, phytic acid and citric acid treatment on the physicochemical and heavy metal adsorption properties of corn starch

Corn starch dispersions were heated with ethanol (E) and reacted with phytic acid (E-PA), citric acid (E-CA), and a mixture of phytic and citric acid (E-PACA) under dry-heating to prepare heavy metal adsorbents. Microscopy images indicated that ethanol treatment induced the formation of porous structures on the surface; furthermore, treatment with phytic and citric acid induced indentations, pores, and irregular structures in E-PA, E-CA, and E-PACA starches. Phytic and citric acid were retained in the starch molecules through ester bonds with the phosphate and carboxyl groups, respectively. Starch esterification by phytic and citric acid induced a loss of crystallinity, high water absorption capacity, and low solubility. E-PACA starch exhibited more efficient Cu2+ adsorption (38.13 mg/g) than native, E, E-PA, and E-CA starches (0.11, 0.49, 2.05, and 36.23 mg/g, respectively). Thus, modification with ethanol, phytic acid and citric acid can be applied to prepare natural starch-based heavy metal adsorbents.

Exacerbating effects of single-dose acute ethanol exposure on neuroinflammation and amelioration by GPR110 (ADGRF1) activation

BackgroundNeuroinflammation is a widely studied phenomenon underlying various neurodegenerative diseases. Earlier study demonstrated that pharmacological activation of GPR110 in both central and peripheral immune cells cooperatively ameliorates neuroinflammation caused by systemic lipopolysaccharide (LPS) administration. Ethanol consumption has been associated with exacerbation of neurodegenerative and systemic inflammatory conditions. The goal of this study is to determine the effects of single-dose acute ethanol exposure and GPR110 activation on the neuro-inflammation mechanisms.MethodsFor in vivo studies, GPR110 wild type (WT) and knockout (KO) mice at 10–12 weeks of age were given an oral gavage of ethanol (3 g/kg) or maltose (5.4 g/kg) at 1–4 h prior to the injection of LPS (1 mg/kg, i.p.) followed by the GPR110 ligand, synaptamide (5 mg/kg). After 2–24 h, brains were collected for the analysis of gene expression by RT-PCR or protein expression by western blotting and enzyme-linked immunosorbent assay (ELISA). Microglial activation was assessed by western blotting and immunohistochemistry. For in vitro studies, microglia and peritoneal macrophages were isolated from adult WT mice and treated with 25 mM ethanol for 4 h and then with LPS (100 ng/ml) followed by 10 nM synaptamide for 2 h for gene expression and 12 h for protein analysis.ResultsSingle-dose exposure to ethanol by gavage before LPS injection upregulated pro-inflammatory cytokine expression in the brain and plasma. The LPS-induced Iba-1 expression in the brain was significantly higher after ethanol pretreatment in both WT and GPR110KO mice. GPR110 ligand decreased the mRNA and/or protein expression of these cytokines and Iba-1 in the WT but not in GPR110KO mice. In the isolated microglia and peritoneal macrophages, ethanol also exacerbated the LPS-induced expression of pro-inflammatory cytokines which was mitigated at least partially by synaptamide. The expression of an inflammasome marker NLRP3 upregulated by LPS was further elevated with prior exposure to ethanol, especially in the brains of GPR110KO mice. Both ethanol and LPS reduced adenylate cyclase 8 mRNA expression which was reversed by the activation of GPR110. PDE4B expression at both mRNA and protein level in the brain increased after ethanol and LPS treatment while synaptamide suppressed its expression in a GPR110-dependent manner.ConclusionSingle-dose ethanol exposure exacerbated LPS-induced inflammatory responses. The GPR110 ligand synaptamide ameliorated this effect of ethanol by counteracting on the cAMP system, the common target for synaptamide and ethanol, and by regulating NLRP3 inflammasome.

Ethanol inhibits pancreatic acinar cell autophagy through upregulation of ATG4B, mediating pathological responses of alcoholic pancreatitis.

Excessive alcohol intake is a major risk factor for pancreatitis, sensitizing the exocrine pancreas to stressors by mechanisms that remain obscure. Impaired autophagy drives nonalcoholic pancreatitis, but the effects of ethanol (EtOH) and alcoholic pancreatitis on autophagy are poorly understood. Here, we find that ethanol reduces autophagosome formation in pancreatic acinar cells, both in a mouse model of alcoholic pancreatitis induced by a combination of EtOH diet and cerulein (a CCK ortholog) and in EtOH+CCK-treated acinar cells (ex vivo model). Ethanol treatments decreased pancreatic level of LC3-II, a key mediator of autophagosome formation. This was caused by ethanol-induced upregulation of ATG4B, a cysteine protease that, cell dependently, regulates the balance between cytosolic LC3-I and membrane-bound LC3-II. We show that ATG4B negatively regulates LC3-II in acinar cells subjected to EtOH treatments. Ethanol raised ATG4B level by inhibiting its degradation, enhanced ATG4B enzymatic activity, and strengthened its interaction with LC3-II. We also found an increase in ATG4B and impaired autophagy in a dissimilar, nonsecretagogue model of alcoholic pancreatitis induced by EtOH plus palmitoleic acid. Adenoviral ATG4B overexpression in acinar cells greatly reduced LC3-II and inhibited autophagy. Furthermore, it aggravated trypsinogen activation and necrosis, mimicking key responses of ex vivo alcoholic pancreatitis. Conversely, shRNA Atg4B knockdown enhanced autophagosome formation and alleviated ethanol-induced acinar cell damage. The results reveal a novel mechanism, whereby ethanol inhibits autophagosome formation and thus sensitizes pancreatitis, and a key role of ATG4B in ethanol's effects on autophagy. Enhancing pancreatic autophagy, particularly by downregulating ATG4B, could be beneficial in mitigating the severity of alcoholic pancreatitis.NEW & NOTEWORTHY Ethanol sensitizes mice and humans to pancreatitis, but the underlying mechanisms remain obscure. Autophagy is important for maintaining pancreatic acinar cell homeostasis, and its impairment drives pancreatitis. This study reveals a novel mechanism, whereby ethanol inhibits autophagosome formation through upregulating ATG4B, a key cysteine protease. ATG4B upregulation inhibits autophagy in acinar cells and aggravates pathological responses of experimental alcoholic pancreatitis. Enhancing pancreatic autophagy, particularly by down-regulating ATG4B, could be beneficial for treatment of alcoholic pancreatitis.

Ethanol inhibition of undifferentiated rat neural progenitor cell replication can be prevented by chlorogenic acid via the NFATc4/CSE signaling pathway.

Prenatal ethanol exposure hinders oxidative stress-mediated neuroblast/neural progenitor cell proliferation by inhibiting G1-S transition, a process vital to neocortical development. We previously showed that ethanol elicits this redox imbalance by repressing cystathionine γ-lyase (CSE), the rate-limiting enzyme in the transsulfuration pathway in fetal brain and cultured cerebral cortical neurons. However, the mechanism by which ethanol impacts the CSE pathway in proliferating neuroblasts is not known. We conducted experiments to define the effects of ethanol on CSE regulation and the molecular signaling events that control this vital pathway. This enabled us to develop an intervention to prevent the ethanol-associated cytostasis. Spontaneously immortalized undifferentiated E18 rat neuroblasts from brain cerebral cortex were exposed to ethanol to mimic an acute consumption pattern in humans. We performed loss- and gain-of-function studies to evaluate whether NFATc4 is a transcriptional regulator of CSE. The neuroprotective effects of chlorogenic acid (CGA) against the effects of ethanol were assessed using ROS and GSH/GSSG assays as measures of oxidative stress, transcriptional activation of NFATc4, and expression of NFATc4 and CSE by qRT-PCR and immunoblotting. Ethanol treatment of E18-neuroblast cells elicited oxidative stress and significantly reduced CSE expression with a concomitant decrease in NFATc4 transcriptional activation and expression. In parallel, inhibition of the calcineurin/NFAT pathway by FK506 exaggerated ethanol-induced CSE loss. In contrast, NFATc4 overexpression prevented loss of ethanol-induced CSE. CGA increased and activated NFATc4, amplified CSE expression, rescued ethanol-induced oxidative stress, and averted the cytostasis of neuroblasts by rescuing cyclin D1 expression. These findings demonstrate that ethanol can perturb CSE-dependent redox homeostasis by impairing the NFATc4 signaling pathway in neuroblasts. Notably, ethanol-associated impairments were rescued by genetic or pharmacological activation of NFATc4. Furthermore, we found a potential role for CGA in mitigating the ethanol-related neuroblast toxicity with a compelling connection to the NFATc4/CSE pathway.

Liposome-assisted penetration and antiaging effects of collagen in a 3D skin model.

Collagen is a major component of the extracellular matrix that supports the epidermal layers of the skin; thus, many strategies have been made to enhance the topical delivery of collagen for antiaging purposes. In addition, our previous study indicated that liposome can help the penetration of active ingredients into the skin. To produce stable collagen-encapsulated liposomes to improve the topical delivery of collagen. Collagen-encapsulated liposomes were fabricated using high-pressure homogenization method. The colloidal stability and adhesion ability were confirmed using dynamic light scattering, and spectrofluorophotometer, respectively. Keratinocyte differentiations of 3D skin before and after treatment with collagen-encapsulated liposomes were confirmed by real-time PCR. In comparison with native collagen, collagen-encapsulated liposomes enhanced collagen retention in artificial membranes by twofold, even after repeated washings with water. In addition, real-time PCR results indicated that 3D skin treated with collagen-encapsulated liposomes exhibited higher levels of collagen, keratin, and involucrin, even after ethanol treatment. Liposomes could serve as efficient delivery vehicles for collagen, thereby enhancing its antiaging effects.