Long-term Heavy Alcohol Consumption Research Articles

Alleviation of Alcoholic Fatty Liver by Dendrobium officinale Flower Extracts due to Regulation of Gut Microbiota and Short-Chain Fatty Acids in Mice Exposed to Chronic Alcohol.

Alcoholic fatty liver disease (AFLD) is caused by long-term heavy alcohol consumption; therefore, useful and practical methods for the prevention of AFLD are urgently needed. The edible flower of Dendrobium officinale contains diverse flavonoids, and has shown antioxidant activity as well as antihypertensive and anti-inflammatory effects. In this study, an AFLD model was established, the protective effect of D. officinale flower (DOF) ethanol extract on AFLD was evaluated, and its mechanisms were investigated by analyzing gut microbiota and short-chain fatty acids (SCFAs). DOF extract (DOFE) supplementation promoted alcohol metabolism, restored hepatic antioxidant capacity, alleviated oxidative stress, reduced inflammatory factor levels, and inhibited dyslipidemia induced by alcohol intake in chronic alcohol-exposed mice, especially in the high DOFE group. Moreover, DOFE supplementation increased the diversity, structure, and composition of the gut microbiota in mice, restored some of the abnormal SCFA levels caused by AFLD, and helped restore intestinal function. DOFE supplementation significantly increased the relative abundance of Akkermansia, suggesting that Akkermansia may be a potential target of the protective effect of DOFE. Therefore, DOFE supplementation to improve the composition of the gut microbiota may be an effective therapeutic strategy for the prevention of AFLD.

The burden of alcoholic cardiomyopathy in China and different regions around the world.

BackgroundAlcoholic cardiomyopathy (ACM) remains a significant public health issue with a growing global burden. The burden of ACM in China and different regions remains poorly understood.MethodsData on ACM deaths, disability-adjusted life years (DALYs), the corresponding global age-standardized death rate (ASDR), age-standardized DALY rate and estimated annual percentage change (EAPC) were analysed based on age, sex, socio-demographic index (SDI) quintiles, different regions and in China from the Global Burden of Disease (GBD) study 2019.ResultsGlobally, the death rate and DALYs due to ACM were 71 723 and 2 441 108 in 2019, 33.06% and 38.79% increase from 1990, respectively. The corresponding ASDR and age-standardized DALY rate decreased with EAPC of -1.52 (95% uncertainty interval (UI) = -2.39, -0.65) and -1.12 (95% UI = -2.14, -0.10). The high-middle SDI regions, especially Eastern Europe, showed the highest number of ACM-related deaths and DALYs. The ACM-related deaths and DALYs were 2545 and 87823 in China in 2019, 171.03% and 147.17% increase from 1990, respectively. Unlike the world level, ASDR and age-standardized DALY rate also increased in China. The ACM burden is higher in men, and people with 50 to 69 years old accounted for the most.ConclusionsACM burden in China and across the world increased substantially from 1990 to 2019. The greatest burden was borne by the high-middle SDI regions, especially by men aged 50-69 years old. Geographically and gender-age tailored strategies were needed to prevent ACM.

Association of Common Variant Rs2281135 of PNPLA3 with Alcohol-Related Cirrhosis in Chinese Han Males

Background and Aim: Alcoholic Liver Disease (ALD), caused by longterm heavy alcohol consumption, is influenced by genetic factors. Studies have illustrated the overlapping genetic mechanism in Nonalcoholic Fatty Liver Disease (NAFLD) and ALD. Recently, a number of Genome-Wide Association Studies (GWAS) have demonstrated several SNPs were strongly associated with NAFLD. The aim of present study is to evaluate the association between these NAFLD-associated SNPs and ALD in Chinese Han population. Methods: Nine SNPs were selected and genotyped in a cohort of 507 patients with ALD and 645 healthy controls by using MassARRAY iPLEX system. Alleles and genotypes analysis of SNPs were performed in logistic regression. The association between SNP and the level of liver serum biomarkers was tested in chi-square test and linear regression model. Results: Our data confirmed that rs2281135 A-allele in PNPLA3 and rs3761472 G-allele in SAMM50 were significantly associated with increased risk of ALD (P = 1.93×10-12, OR [95% CI] = 1.82 [1.54-2.15]; P = 2.08×10-16, OR [95% CI] = 1.06 [1.04-1.08], respectively). The genotypes of rs2281135 were associated with ALD in additive, dominant and recessive genetic model (P = 1.24×10-11, P = 1.46×10-7, P = 2.07×10-9, respectively). In addition, rs2281135 was found to be associated with serum elevated levels of ALT (P = 5.0×10-3), AST (P = 0.03), ALP (P = 0.02), GGT (P = 0.03) in patients with ALD. Conclusions: The present study confirmed that PNPLA3 common variant rs2281135 was significantly associated with ALD in Chinese male Han population.

Clinical impact of acquired risk factors in TTN-mediated dilated cardiomyopathy

Abstract Background/Introduction Truncating variants in TTN-encoded titin (TTNtvs) underlie ∼25% of familial dilated cardiomyopathy (DCM) cases. In addition, TTNtvs are over-represented in alcoholic, peripartum, and chemotherapy-induced CM suggesting that risk factors may play a substantive role in the penetrance and expressivity of TTN-mediated DCM. Purpose To determine the prevalence and clinical impact of risk factors for TTN-mediated DCM within a single center cohort of unrelated patients with pathogenic (P)/likely pathogenic (LP) TTNtvs. Methods In this retrospective study, an institutionally developed natural language processing algorithm was used to identify all TTNtv-positive patients evaluated clinically between 01/2012 and 12/2019. Each TTNtv was then mapped to the TTN reference sequence (NM_133378.4) and classified according to a strict interpretation of the 2015 American College of Medical Genetics and Genomics (ACMG) guidelines. After exclusion of individuals with complex congenital heart disease, ischemic heart disease, and those without ACMG P/LP TTNtvs, available medical records were reviewed for risk factors such as long-term heavy alcohol consumption (>80 grams/day), cardiotoxic chemotherapy exposure, morbid obesity (body mass index >40), biopsy/imaging-confirmed myocarditis, peripartum/tachycardia-mediated CM, and valvular heart disease. Results Overall, 33 ACMG P/LP TTNtv-positive patients were identified (40% female; mean age at presentation 45±16 years; mean LVEF at presentation 38%±17%). Of note, 17/33 (52%) presented with heart failure (HF) and 4/33 (12%) presented with sustained ventricular arrhythmias (VAs). The remaining 12 (36%) TTNtv-positive patients presented with incidentally detected asymptomatic left ventricular systolic dysfunction (n=9) or as a result of genetic cascade screening (n=3). In comparison to TTNtv-positive patients with non-HF presentations, those with HF had worse left ventricular function (mean LVEF: 26% ± 11% vs 51% ±12%; p<0.001) and were more likely to have ≥1 risk factor [11/17 (65%) vs 4/16 (25%); p=0.04]. Unexpectedly, the 7 TTNtv-positive patients that presented with (n=4) or subsequently developed (n=3) sustained VAs were younger at the time of presentation (34±13 years vs 48±15 years; p=0.04) and displayed a similar degree of left ventricular dysfunction to those without sustained VAs (mean LVEF: 42% ± 17% vs 36% ± 17%; p=ns). Interestingly, the only risk factor over-represented amongst those with sustained VAs was bileaflet mitral valve prolapse [3/7 (43%) vs 0/26 (0%); p=0.006]. Conclusion The majority of TTNtv-positive patients (14/23; 61%) with a history of HF and/or sustained VAs possessed ≥1 acquired risk factor providing further evidence that gene-environment interaction(s) exert a crucial role in the manifestation of TTN-mediated disease. Funding Acknowledgement Type of funding source: None

Alcohol Inhibits Odontogenic Differentiation of Human Dental Pulp Cells by Activating mTOR Signaling

Long-term heavy alcohol consumption could result in a range of health, social, and behavioral problems. People who abuse alcohol are at high risks of seriously having osteopenia, periodontal disease, and compromised oral health. However, the role of ethanol (EtOH) in the biological functions of human dental pulp cells (DPCs) is unknown. Whether EtOH affects the odontoblastic differentiation of DPCs through the mechanistic target of rapamycin (mTOR) remains unexplored. The objective of this study was to investigate the effects of EtOH on DPC differentiation and mineralization. DPCs were isolated and purified from human dental pulps. The proliferation and odontoblastic differentiation of DPCs treated with EtOH were subsequently investigated. Different doses of EtOH were shown to be cytocompatible with DPCs. EtOH significantly activated the mTOR pathway in a dose-dependent manner. In addition, EtOH downregulated the alkaline phosphatase activity, attenuated the mineralized nodule formation, and suppressed the expression of odontoblastic markers including ALP, DSPP, DMP-1, Runx2, and OCN. Moreover, the pretreatment with rapamycin, a specific mTOR inhibitor, markedly reversed the EtOH-induced odontoblastic differentiation and cell mineralization. Our findings show for the first time that EtOH can suppress DPC differentiation and mineralization in a mTOR-dependent manner, indicating that EtOH may be involved in negatively regulating the dental pulp repair.

Targeting Pin1 Protects Mouse Cardiomyocytes from High-Dose Alcohol-Induced Apoptosis.

Long-term heavy alcohol consumption is considered to be one of the main causes of left ventricular dysfunction in alcoholic cardiomyopathy (ACM). As previously suggested, high-dose alcohol induces oxidation stress and apoptosis of cardiomyocytes. However, the underlying mechanisms are yet to be elucidated. In this study, we found that high-dose alcohol treatment stimulated expression and activity of Pin1 in mouse primary cardiomyocytes. While siRNA-mediated knockdown of Pin1 suppressed alcohol-induced mouse cardiomyocyte apoptosis, overexpression of Pin1 further upregulated the numbers of apoptotic mouse cardiomyocytes. We further demonstrated that Pin1 promotes mitochondria oxidative stress and loss of mitochondrial membrane potential but suppresses endothelial nitric oxide synthase (eNOS) expression in the presence of alcohol. Taken together, our results revealed a pivotal role of Pin1 in regulation of alcohol-induced mouse cardiomyocytes apoptosis by promoting reactive oxygen species (ROS) accumulation and repressing eNOS expression, which could be potential therapeutic targets for ACM.

T2 relaxation time alterations underlying neurocognitive deficits in alcohol-use disorders (AUD) in an Indian population: A combined conventional ROI and voxel-based relaxometry analysis

Long-term heavy alcohol consumption has traditionally been associated with impaired cognitive abilities, such as deficits in abstract reasoning, problem solving, verbal fluency, memory, attention, and visuospatial processing. The present study aimed at exploring these neuropsychological deficits in alcohol-use disorders (AUD) in an Indian population using the Postgraduate Institute Battery of Brain Dysfunction (PGIBBD) and their possible correlation with alterations in T2 relaxation times (T2-RT), using whole-brain voxel-based relaxometry (VBR) and conventional region of interest (ROI) approach. Multi-echo T2 mapping sequence was performed on 25 subjects with AUD and 25 healthy controls matched for age, education, and socioeconomic status. Whole-brain T2-RT measurements were conducted using VBR and conventional ROI approach. The study was carried out on a 3T whole-body MR scanner. Post processing for VBR and ROI analysis was performed using SPM 8 software and vendor-provided software, respectively. A PGIBBD test battery was conducted on all subjects to assess their cognitive abilities, and the results were reported as raw scores. VBR and ROI results revealed that AUD subjects showed prolonged T2-RTs in cerebellum bilaterally, parahippocampal gyrus bilaterally, right anterior cingulate cortex, left superior temporal gyrus, left middle frontal gyrus, and left calcarine gyrus. A significant correlation was also observed between the neuropsychological test raw scores and alterations in T2-RT in AUD subjects. Our results are consistent with previous studies suggesting tissue disruption or gliosis or demyelination as a possible reason for prolonged T2-RTs. This damage to brain tissue, which is evident as prolonged T2-RT, could possibly be associated with impaired cognitive abilities noticeable in AUD subjects.

Abstract 14641: Deficiency in the E3 Ubiquitin Ligase Parkin Exacerbates Alcoholic Cardiomyopathy: Role of Mitophagy

Background: Long-term heavy alcohol consumption has been shown to promote mitochondrial injury, unfavorable geometric and contractile changes in the heart. Parkin, a cytosolic E3 ubiquitin ligase encoded by PARK2 gene, plays an important role in the regulation of selective mitophagy. This study was designed to examine the role of Parkin in alcohol-induced myocardial injury (aka alcoholic cardiomyopathy) and the underlying mechanism with a focus on mitophagy. Methods: Adult male wild-type C57 and PARKIN2 knockout (Parkin-/-) mice were placed on alcohol (4%) or control diet for 4 weeks. Echocardiographic and cardiomyocyte mechanical properties were assessed. Mitochondrial morphology, function and mitophagy were examined using transmission electronic microscopy, Clark-type oxygen electrode, and Western blot, respectively. Results: Our results revealed that chronic alcohol consumption triggered unfavorable geometric and contractile changes [decreased fractional shortening (FS) and ejection fraction (EF), with enlarged left ventricular chamber; decreased peak shortening (PS) and velocity of shortening +dL/dt, increased time-to-90% relengthening TR90], the effects of which were exacerbated by Parkin deficiency. In addition, our data showed that chronic alcohol intake promoted myocardial mitochondrial swelling with cristae disarrangement, induced myocardial mitochondrial depolarization and respiration inhibition, which were exacerbated by Parkin knockout. Furthermore, chronic alcohol consumption promoted mitophagy activation, as evidenced by accumulation of Parkin and LC3BII in mitochondria and mitochondrial ubiquitination level in the heart, the effect of which was nullified by Parkin knockout. Conclusion: These data suggest that chronic alcohol consumption triggered mitophagy by stimulating Parkin translocation to the mitochondria, which may be an adaptive response in the heart. Our findings implicated the therapeutic potential of mitophagy as a target in the management of alcoholic cardiomyopathy.

Alcoholic cardiomyopathy: pathophysiologic insights.

Alcoholic cardiomyopathy (ACM) is a specific heart muscle disease found in individuals with a history of long-term heavy alcohol consumption. ACM is associated with a number of adverse histological, cellular, and structural changes within the myocardium. Several mechanisms are implicated in mediating the adverse effects of ethanol, including the generation of oxidative stress, apoptotic cell death, impaired mitochondrial bioenergetics/stress, derangements in fatty acid metabolism and transport, and accelerated protein catabolism. In this review, we discuss the evidence for such mechanisms and present the potential importance of drinking patterns, genetic susceptibility, nutritional factors, race, and sex. The purpose of this review is to provide a mechanistic paradigm for future research in the area of ACM.

Regulation of pancreatic inflammation by connective tissue growth factor (CTGF/CCN2).

Pancreatitis is caused by long-term heavy alcohol consumption, which results in injury and death of pancreatic acinar cells (PAC). The PAC play a pivotal role in mediating early inflammatory responses but the underlying mechanisms remain poorly understood. Treatment of C57BL/6 mice with ethanol and cerulein resulted in increased staining for acinar interleukin- 1b (IL-1b), chemokine (C-C motif) ligand 3 (CCL3), or connective tissue growth factor (CTGF/CCN2) by Day 16 and this was associated with increased infiltration of F4/80-positive macrophages and increased expression of pancreatic CTGF/CCN2 mRNA. Compared with wild-type Swiss Webster mice, ethanol treatment of pan-green fluorescent protein (GFP)-CTGF/CCN2 transgenic mice caused enhanced acinar staining for GFP or CTGF/CCN2 and a significant increase in pancreatic infiltration of F4/80-positive macrophages or NIMP-R14-positive neutrophils. Treatment of primary mouse PAC or the rat AR42J PAC line with ethanol or CTGF/CCN2 resulted in enhanced expression of IL-1b or CCL3. Conditioned medium from CTGF/CCN2-treated AR42J cells induced chemotaxis in NR8383 macrophages and this response was abrogated in a dose dependent manner by addition of BX471, an inhibitor of chemokine (C-C motif) receptor 1. These results reveal that acinar CTGF/CCN2 plays a novel role in alcohol-induced inflammatory processes in the pancreas by increasing infiltration of macrophages and neutrophils and increasing acinar production of inflammatory mediators such as IL-1b or CCL3. The early production of CTGF/CCN2 by PAC to drive inflammation is distinct from its previously reported production by pancreatic stellate cells to drive fibrosis at later stages of pancreatic injury.

Value of Fibrosis Markers for Staging Liver Fibrosis in Patients With Precirrhotic Alcoholic Liver Disease

Our aim was to identify markers predictive of fibrosis in alcoholic liver disease (ALD). Percutaneous liver biopsy is the recommended standard for histologic assessment of liver fibrosis. Seven serum markers (tissue inhibitor of matrix metalloproteinase 1 [TIMP1], tenascin, collagen VI, amino-terminal propeptide of type III collagen [PIIINP], matrix metalloproteinases [MMP2], laminin, and hyaluronic acid [HA]) representing various aspects of collagen and extracellular matrix deposition and degradation, have been proposed as noninvasive surrogates for liver biopsy. Moreover, a diagnostic algorithm including 3 serum markers (TIMP1, PIIINP, HA) and age has been proposed to accurately detect fibrosis with acceptable levels of sensitivity/specificity in a chronic hepatitis C subgroup. To determine variability of these markers in liver fibrosis with different etiologies, we conducted an evaluation of their correlative properties in a subgroup of patients (n = 247) with biopsy confirmed liver fibrosis resulting from long-term heavy alcohol consumption. Patients were participants in a recently completed VA multicenter clinical trial followed over 2 years with liver biopsy at baseline and 24 months, and with markers assessed every 3 months. Among the markers measured in this alcoholic subgroup all except collagen VI displayed significant correlation with degrees of fibrosis. Three markers, TIMP1, PIIINP and HA adjusted for age, emerged as the most promising predictors of the degree of fibrosis in a population of alcoholics. However, there was little change over time as related to change in fibrosis. The lower than expected accuracy of these markers based on receiver operating curves (ROC) also showed their limited use in this etiologic subgroup. In alcoholic patients, various markers have limited value in predicting and diagnosing the stages of fibrosis compared to liver biopsy. Thus, further prospective studies are required to better define the usefulness of each marker or their combination which are possibly affected by alcohol metabolism.

Response shifting and inhibition, but not working memory, are impaired after long-term heavy alcohol consumption.

Chronic alcohol abuse leads to cognitive deficits. The authors investigated whether a systematic increase of interference in a 2-back working memory paradigm would lead to cognitive deficits in alcoholic participants and compared their performance in such a task with that in an alternate-response task. Twenty-four nonamnesic and nondemented alcohol abuse (AA) patients and 12 patients with Korsakoff's syndrome (KS) were compared with a control group. AA patients were impaired in the alternate-response task but not in working memory interference resolution. KS patients performed worse than the AA patients and the controls in both tasks. The neurotoxic side effects of alcohol therefore lead to a specific deficit in alternating between response rules but not in working memory, independently of whether the working memory task involves interference resolution or not.

Frontal lobe dysfunctions in Korsakoff's syndrome and chronic alcoholism: continuity or discontinuity?

The effect of long-term heavy alcohol consumption on brain functions is still under debate. The authors investigated a sample of 17 Korsakoff amnesics, 23 alcoholics without Korsakoff's syndrome, and 21 controls with peripheral nerve diseases, matched for intelligence and education. Executive functions were examined for word fluency, the modified Wisconsin Card Sorting Test, an alternate response task, and an "n-back" working memory task. Korsakoff amnesics, but not alcoholics, showed a marked memory impairment. They also scored lower in each of the executive tasks-the alcoholics only in the alternate response task. This task also correlated with the years of the alcohol dependency. First, the authors conclude that Korsakoff's syndrome is associated not only with a memory impairment but also with a global executive deficit. Second, the decline in the ability to alternate between different responses argues for a restricted neurotoxic effect of alcohol on some frontal lobe areas.

Is long-term heavy alcohol consumption toxic for brain serotonergic neurons? Relationship between years of excessive alcohol consumption and serotonergic neurotransmission

The relationship between years of excessive alcohol consumption and central serotonergic neurotransmission, as assessed by the prolactin (PRL) response to d-fenfluramine, was investigated in 22 male alcohol-dependent subjects. A negative correlation was obtained, that is, the longer duration of excessive alcohol consumption the lower PRL response to d-fenfluramine. It is therefore suggested that long duration of excessive alcohol consumption in alcohol-dependent subjects causes a reduction in central serotonergic neurotransmission, possibly by a toxic effect of alcohol on serotonin neurons. The relationship between depressive and anxiety symptoms during on-going drinking and the PRL response to d-fenfluramine was also investigated. No such correlations were obtained, suggesting that reduction in central serotonergic neurotransmission does not pre-dispose to the development of depressive and anxiety symptoms, at least in relation to on-going drinking in alcohol-dependent subjects.

Risk factors for stroke due to cerebral infarction in young adults.

Stroke in the young is particularly tragic because of the potential for a lifetime of disablement. More than 10% of patients with stroke due to cerebral infarction are aged 55 years or younger. While a number of studies have addressed the issue of stroke mechanism in the young, quantitation of risk factors has rarely been undertaken. Given the importance of risk factor assessment in primary prevention, we aimed to assess this using case-control methodology in a hospital-based series and community-based control subjects. A total of 201 consecutive patients with first-onset stroke due to cerebral infarction aged 15 to 55 years (mean, 45.5 years) were accrued from four teaching hospitals during 1985 to 1992 and compared with their age- and sex-matched neighborhood controls. Information concerning potential risk factor exposure status was collected by structured questionnaire at interview. Stroke risks were estimated by calculating the odds ratios with multivariate logistic regression. Significantly increased risk of stroke was found among those with diabetes (odds ratio, 11.6 [95% confidence intervals, 1.2 to 115.2]), hypertension (6.8 [3.3 to 13.9]), heart disease (2.7 [1.1 to 6.4]), current cigarette smoking (2.5 [1.3, 5.0]), and long-term heavy alcohol consumption (> or = 60 g/d) (15.3 [1.0 to 232.0]). However, heavy alcohol ingestion (> or = 60 g) within 24 hours preceding stroke onset was not a risk factor (0.9 [0.3 to 3.4]). Diabetes, hypertension, heart disease, current smoking, and long-term heavy alcohol consumption are major risk factors for stroke in young adults. Given that the majority of these factors are either correctable or modifiable, prevention strategies may have the potential to reduce the impact of stroke in this age group.