Development Of Nonalcoholic Fatty Liver Disease Research Articles

Application and mechanism of Chinese herb medicine in the treatment of non-alcoholic fatty liver disease

Non-alcoholic fatty liver disease (NAFLD) is a chronic liver condition closely associated with metabolic syndrome, with its incidence rate continuously rising globally. Recent studies have shown that the development of NAFLD is associated with insulin resistance, lipid metabolism disorder, oxidative stress and endoplasmic reticulum stress. Therapeutic strategies for NAFLD include lifestyle modifications, pharmacological treatments, and emerging biological therapies; however, there is currently no specific drug to treat NAFLD. However Chinese herb medicine (CHM) has shown potential in the treatment of NAFLD due to its unique therapeutic concepts and methods for centuries in China. This review aims to summarize the pathogenesis of NAFLD and some CHMs that have been shown to have therapeutic effects on NAFLD, thus enriching the scientific connotation of TCM theories and facilitating the exploration of TCM in the treatment of NAFLD.

TLR4 AS THE MARKER OF NON-ALCOHOLIC FATTY LIVER DISEASE PROGNOSIS IN PATIENTS WITH INTESTINAL DYSBIOSIS

Introduction. Non-alcoholic fatty liver disease (NAFLD) is one of the most common liver diseases characterized by the fat accumulation in liver cells and is closely associated with metabolic disorders including obesity and insulin resistance. Considerable interest of researchers is focused on the role of the intestinal microbiome in the NAFLD development and occurrence, since a violation of the normal bacteria balance (dysbiosis) can increase inflammatory processes in the liver. Toll-like receptor 4 (TLR4), which responds to components of the bacterial membrane, is an important marker involved in the development of inflammation and progression of NAFLD.The aim of this study was to determine the level of TLR4 in patients with NAFLD and to find the relationship with gut microbiome components. The study included 152 patients diagnosed with NAFLD and 86 control patients without fatty liver infiltration. General clinical, biochemical, enzyme immunoassay, instrumental and bacteriological methods were used to assess the condition of patients.The results showed that the level of TLR4 in patients with NAFLD was significantly higher than in the control group (2.67±3.05 ng/ml vs. 1.23±0.99 ng/ml). A positive correlation was marked between the TLR4 level and biochemical indicators of inflammation, such as ALT, AST, TG and ALP. It confirms the role of TLR4 in the development of inflammatory processes in NAFLD.In addition, patients with NAFLD showed a significant change in the gut microbiome composition – an increase in the number of Firmicutes phylotype bacteria and a higher level of Firmicutes/Bacteroidetes index, which was in positive correlation with TLR4. Small intestinal bacterial overgrowth (SIBO), which may lead to TLR4 activation, was also marked in 50% of NAFLD patients.Conclusion. The study confirms the role of TLR4 as a marker of the potential tole on NAFLD development and its relationship with intestinal dysbiosis, which could be perspective for the looking of new approaches in the prognosis and treatment of this disease. The microbiome correction and the development of TLR4 inhibitors is an essential way for future investigations as promising therapeutic method.

The effect of ertugliflozin in patients with nonalcoholic fatty liver disease associated with type 2 diabetes mellitus: A randomized controlled trial

Background: Nonalcoholic fatty liver disease (NAFLD) is a chronic liver disease associated with liver inflammation, fibrosis, and cirrhosis and is associated with a greater risk of hepatocarcinoma. Nonalcoholic steatohepatitis (NASH) is a persistent and progressive form of NAFLD. Recent evidence suggested that ertugliflozin, a sodium-glucose cotransporter 2 inhibitor (SGLT2), suppresses NAFLD development in patients with type 2 diabetes mellitus (T2DM). The objective of this study was to determine the impact of ertugliflozin on improving NAFLD in patients with T2DM and the function of liver enzymes. Methods: This prospective, randomized, double-blind, placebo-controlled, interventional study aimed to determine the effectiveness of 15 mg of ertugliflozin versus 30 mg of the standard therapy pioglitazone versus placebo in NAFLD patients with T2DM. The study was established based on patient randomization in three groups: ertugliflozin, pioglitazone, and a placebo. This study was registered under the Australian New Zealand Clinical Trial Registry (Trial ID: ACTRN12624000032550). Results: The impact of therapy was determined in the treatment groups by utilizing liver ultrasonography and biochemical parameters. After 24 weeks of clinical study, the results revealed significant improvement in the grades of fatty liver, especially in the ertugliflozin group. The number of patients with hepatic steatosis significantly decreased among the respective groups classified according to fatty liver grade. Among patients in the ertugliflozin and pioglitazone groups, 45% to 23.4% and 41.7% to 26.6%, respectively, decreased in the Grade 2 group. The aspartate aminotransferase and alanine aminotransferase levels were significantly lower in all the study groups, especially in the ertugliflozin group (P ≤ .001). Conclusion: The present study revealed that the concomitant use of ertugliflozin has favorable effects on liver enzymes, as it decreases liver fat intake and reduces complications in patients with NAFLD-associated T2DM. However, more in-depth studies will be required to observe every aspect of ertugliflozin.

Helicobacter Pylori Infection Is not Associated with Nonalcoholic Fatty Liver Disease: A Two-year Cohort Study.

Previous studies reported inconsistent results of the association between Helicobacter pylori (H. pylori) infection and nonalcoholic fatty liver disease (NAFLD). A cohort study of 2063 adults without NAFLD at baseline, who participated in a repeated health check-up including a 13C urea breath test and abdominal ultrasonography, was conducted to evaluate the link between H. pylori infection and NAFLD development. During a mean follow-up period of 1.7 years, we did not found a significant association between H. pylori infection and NAFLD (Hazard ratio (HR) = 1.10 (0.86, 1.40), p = 0.4689). We also found that higher age, body mass index (BMI), systolic blood pressure (systolic BP), diastolic blood pressure (diastolic BP), fasting blood glucose, triglycerides, total cholesterol, low density lipoprotein cholesterol (LDL-C), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were risk factors, and high density lipoprotein cholesterol (HDL-C) was a protective factor for NAFLD development. H. pylori infection might not be positively related to NAFLD development.

Association between nutritional status indices and non-alcoholic fatty liver disease in older adults: insights from the National Health and Nutrition Examination Survey 2017-2018.

While previous studies have identified a relationship between dietary intake and the risk of non-alcoholic fatty liver disease (NAFLD), the influence of overall nutritional status on NAFLD development has not been thoroughly investigated. This study sought to explore the association between different nutritional status indicators and NAFLD among the older adults. Nutritional status was evaluated using controlling nutritional status (CONUT), prognostic nutritional index (PNI) and nutritional risk index (GNRI), while NAFLD was identified based on a controlled attenuation parameter ≥ 285 dB/m, measured using transient elastography. The analysis included multivariate regression, receiver operating characteristic analysis, eXtreme Gradient Boosting and subgroup analysis to investigate the relationships between nutritional status indices and NAFLD. The study enrolled 1409 participants for the main analysis, with an NAFLD prevalence of 44·7 %. After accounting for potential confounders, a positive association between PNI and NAFLD was observed. Participants in the third and fourth quartiles of PNI showed increased odds of NAFLD compared with the lowest quartile (Q3: OR = 1·45, 95 % CI (1·03, 2·05); Q4: OR = 2·27, 95 % CI (1·59, 3·24)). Similarly, higher GNRI quartiles were significantly associated with greater odds of NAFLD (Q4 v. Q1: aOR = 1·84; 95 % CI (1·28, 2·65)). Conversely, higher CONUT values were linked to a reduced prevalence of NAFLD (OR = 0·65, 95 % CI (0·48, 0·87)). This study highlights that suboptimal nutritional status, indicating overnutrition as evaluated by PNI, GNRI and CONUT, is positively linked with the risk of NAFLD in individuals aged 50 years and above.

U-shaped association of serum vitamin A concentrations with all-cause mortality in patients with NAFLD: results from the NHANES database prospective cohort study

BackgroundPrevious studies have demonstrated a significant association between serum vitamin A concentration and non-alcoholic fatty liver disease (NAFLD) development. However, the long-term prognostic implications of serum vitamin A in patients with NAFLD remain underexplored. This study aims to investigate whether there exists a correlation between serum vitamin A concentrations and overall mortality among subjects diagnosed with NAFLD.MethodsTo investigate the association between serum vitamin A concentrations and NAFLD outcomes, we conducted prospective cohort studies using data from the 1999–2006 and 2017–2018 National Health and Nutrition Examination Survey (NHANES). We utilized a multivariate Cox regression model to explore the relationship between serum vitamin A levels and all-cause mortality. Survival curves related to serum vitamin A were constructed using the Kaplan–Meier method. Additionally, the restricted cubic splines (RCS) method was applied to examine potential nonlinear relationships between serum vitamin A concentrations and all-cause mortality of NAFLD.ResultsOver a median follow-up period of 10.3 years, a total of 1,399 all-cause deaths were recorded. The weighted average concentration of serum vitamin A was 61.48 ± 0.37 μg/dL. After adjusting for potential confounders, a significant U-shaped relationship was identified between serum vitamin A concentrations and the risk of all-cause mortality in NAFLD patients. This relationship was particularly pronounced in men and elderly individuals aged 60 to 85.ConclusionOur study reveals a significant non-linear relationship between serum vitamin A concentrations and the risk of all-cause mortality in patients with NAFLD. These findings underscore the importance of monitoring and maintaining optimal serum vitamin A levels to potentially improve survival outcomes in NAFLD patients.

Nonalcoholic Fatty Liver Disease Risk and Proprotein Convertase Subtilisin Kexin 9 in Familial Hypercholesterolemia Under Statin Treatment

Background/Objectives: Fatty acids are involved in some hepatic disorders. The proprotein convertase subtilisin kexin 9 (PCSK9) inhibits the uptake of low-density lipoproteins (LDLs), which contain lipids, into the liver and may thus be associated with nonalcoholic fatty liver disease (NAFLD), a cardiovascular disorder (CVD) risk. Statins reduce blood LDL–cholesterol (LDL-C) levels and CVD risk and can attenuate the development of NAFLD while increasing blood PCSK9 levels. Methods: We investigated the correlation between PCSK9 and liver conditions in patients with familial hypercholesterolemia (FH), a CVD risk population with elevated blood LDL-C levels, under statin treatment. Blood tests for lipids, PCSK9, and liver function (aspartate aminotransferase [AST] and alanine aminotransferase [ALT]) were performed in patients with FH taking statins (n = 25, mean age = 57 years, 12% of males). The ALT:AST ratio was used as a marker of NAFLD risk. Results: The mean LDL-C level was 3.38 mmol/L, and the median PCSK9 level was 312 ng/mL. The median ALT:AST ratio was 0.88. A significant negative correlation was observed between the PCSK9 and ALT:AST ratio (β = −0.67, p < 0.05). Conclusions: Their negative correlation might give a hypothetical insight into the effect of statin treatment on the development of NAFLD, in relation to PCSK9 behavior, in patients with FH.

6428 Update on Genetic F6428 actors of Nonalcoholic Fatty Liver Disease

Abstract Disclosure: H.M. Heshmati: None. Background: Nonalcoholic fatty liver disease (NAFLD) is a pandemic with a prevalence of 25% among the adult population worldwide. NAFLD is a spectrum of liver disorders ranging from simple steatosis to nonalcoholic steatohepatitis (NASH). NASH can progress to cirrhosis and hepatocellular cancer. The pathogenesis of NAFLD is complex and multifactorial, involving metabolic, epigenetic, and genetic factors. Several medical conditions (e.g., obesity, type 2 diabetes, inflammation, insulin resistance, disrupted gut microbiome, and impaired intestinal barrier function) are important risk factors associated with and/or contributing to NAFLD. This review presents an update on the genetic factors of NAFLD identified by the genome-wide association studies (GWAS). Methods: A systematic search of literature was conducted using the search terms nonalcoholic fatty liver disease, nonalcoholic steatohepatitis, cirrhosis, hepatocellular cancer, obesity, type 2 diabetes, epigenetics, and genetics. Results: Evidence for the contribution of genetic factors to NAFLD is further emphasized by the ethnic differences in the prevalence of NAFLD (e.g., higher prevalence among Hispanic American compared to African American adults). Based on the results of the GWAS, several genetic modifiers are relevant to NAFLD development and progression. They include variations in patatin-like phospholipase domain-containing protein 3 (PNPLA3), transmembrane 6 superfamily 2 (TM6SF2), membrane-bound O-acyltransferase domain-containing protein 7 (MBOAT7), glucokinase regulatory protein (GCKR), programmed cell death protein 1 (PDCD1), and hydroxysteroid 17β-dehydrogenase 13 (HSD17B13) genes (non-exhaustive list). These genetic modifiers interact with the metabolic and epigenetic factors for the development and progression of NAFLD and are responsible for the variability in the severity of NAFLD. The PNPLA3 gene variant (rs738409), which is the first and the best validated genetic modifier, is associated with a high risk of development and progression of NAFLD. The PDCD1 gene variant (rs7421861) is independently associated with hepatocellular cancer development. In contrast, the HSD17B13 gene variant (rs72613567) may have a protective effect for NAFLD risk. Conclusion: NAFLD is a multifactorial condition with a complex pathogenesis involving the interactions between metabolic, epigenetic, and genetic factors. The prevalence and risk of NAFLD differ across populations mainly due to genetic polymorphisms. Genetic factors relevant to NAFLD include variations in PNPLA3, TM6SF2, MBOAT7, GCKR, PDCD1, and HSD17B13 genes. A better understanding of these genetic modifiers and their mechanisms of action should allow a better risk prediction of the development of NAFLD and its progression to cirrhosis and hepatocellular cancer and help the development of new and selected therapies for NAFLD. Presentation: 6/3/2024

Association between hepatocyte TM4SF5 expression and gut microbiome dysbiosis during non-alcoholic fatty liver disease development

Gut microbiome dysbiosis is involved in non-alcoholic fatty liver disease (NAFLD) development. Hepatic transmembrane 4 L six family member 5 (TM4SF5) overexpression promotes NAFLD. However, how gut microbiota are associated with TM4SF5-mediated NAFLD remains unexplored. We analyzed the gut microbiome using feces from hepatocyte-specific TM4SF5-overexpressing transgenic (Alb-TGTm4sf5-Flag, TG) or Tm4sf5−/− knock-out (KO) mice fed a normal chow diet (NCD), high-fat diet (HFD) for 2 weeks (HFD2W), or methionine-choline-deficient diet (MCD) for 4 weeks to investigate associations among Tm4sf5 expression, diet, and the gut microbiome. TG-NCD mice showed a higher Firmicutes-to-Bacteroidetes (F/B) ratio, with less enrichment of Akkermansia muciniphila and Lactobacillus reuteri. NASH-related microbiomes in feces were more abundant in TG-HFD2w mice than in KO-HFD2w mice. Further, TG-MCD showed a higher F/B ratio than TG-NCD or KO mice, with decreases or increases in microbiomes beneficial or detrimental to the liver, respectively. Such effects in TG-MCD animals were correlated with functional pathways producing short-chain fatty acids (SCFAs). Furthermore, potential functional pathways of the gut microbiome were metabolically parallel to NAFLD features in TG-MCD mice. These results suggest that hepatocyte Tm4sf5 supports gut microbiome dysbiosis and metabolic activity, leading to SCFA production and hepatic inflammation during NAFLD development.

Lygodium microphyllum Inhibits de Novo Lipogenesis Activity in the Hepatocytes of High-Fat High-Fructose-Induced Rats by Increasing the Levels of SIRT1 and AMPK.

The prevalence of non-alcoholic fatty liver disease (NAFLD) is currently of great concern due to its risk of developing T2DM and cardiovascular disease. The development of NAFLD may be initiated by de novo lipogenesis in the hepatocytes. Sirtuin1 (SIRT1) and adenosine monophosphate-activated protein kinase (AMPK), are responsible for the lipogenesis mechanism. Interestingly, plant sterols, such as beta-sitosterol and stigmasterol, have the potential to lower the LDL-cholesterol in dyslipidemic patients. Beta-sitosterol was present in the ethanol extract of Lygodium microphyllum herbs at a concentration of 283.55µg/g extract. This sterol interacted with the active allosteric-binding site of SIRT1 and AMPK similarly to the proteins' activators. To investigate the anti-lipogenesis activity of the ethanol extract of L. microphyllum (ELM) in the liver tissue of rats through the SIRT1 and AMPK levels. Forty male Wistar rats were used in this study: (1) normal control group; (2) high-fat high-fructose diet (HFHFD) rats; (3) HFHFD rats treated with metformin; (4) HFHFD rats treated with resveratrol; (5) HFHFD rats treated with beta-sitosterol; (6-8) HFHFD rats treated with ELM doses of 200, 400, and 600 mg/kg BW. Rats in the normal control group were fed regular chow, while other groups of rats were given HFHFD for 35 days. All drugs were given orally on D15 till D35. On D35, the rats were sacrificed, and the liver organs were examined for the liver index, morphology, NAFLD activity score (NAS), and levels of SIRT1 and AMPK. ELM improves the morphology, the liver index, the steatosis condition, and the NAS of HFHFD-induced NAFLD rats. ELM increases the levels of SIRT1 and AMPK in the liver tissue of HFHFD-induced NAFLD rats. ELM may have the potential to inhibit de novo lipogenesis by increasing the levels of SIRT1 and AMPK.

Hesperitin prevents non-alcoholic steatohepatitis by modulating mitochondrial dynamics and mitophagy via the AMPKα-Drp1/PINK1-Parkin signaling pathway

Non-alcoholic fatty liver disease (NAFLD) is becoming a global public health burden, yet effective therapeutic strategies are notably lacking. NAFLD development may be mediated by mitochondrial dysfunction, according to new research. Producing mitochondrial regulators from plant-based substances to treat mitochondrial dysfunction is an appealing approach to treating NAFLD. Hesperetin (HES) is a flavonoid that is found naturally and is a member of the flavanone family. This study aims to clarify the mechanism of HES in preventing NAFLD which is caused by a high-fat diet (HFD). Serum and liver biochemical parameters, liver histology, lipid profiles, and mitochondrial function were evaluated in HFD-induced NAFLD Sprague-Dawley (SD) rats. HES treatment significantly reduced body weight gain, liver weight, and the liver index, while also improving hepatic steatosis, lipid metabolism disorders, and mitochondrial dysfunction in rats with NAFLD. The mechanism was investigated and confirmed using western blot and real-time quantitative polymerase chain reaction (RT-qPCR). We showed that in the liver of NAFLD rats, HES decreased the expression of dynamic-related protein 1 (Drp1), phosphorylated Drp1 at serine-616 (Drp1-pS616) and induced phosphorylated Drp1 at serine-637 (Drp1-pS637), PTEN-induced kinase 1 (PINK1), and E3 Ubiquitin-Protein Ligase Parkin (Parkin) via an AMP-activated protein kinase alpha (AMPKα)-dependent mechanism. Moreover, HES increased the expression of the mitochondrial fusion proteins mitofusin-2 (Mfn2) and optic atrophy 1 (Opa1) while suppressing the expression of fission protein 1 (Fis1). In this work, we identify a unique mechanism by which HES prevents NAFLD from developing. HES may be an attractive potential therapeutic agent to cure NAFLD.

Branched-chain amino acids alleviate NAFLD via inhibiting de novo lipogenesis and activating fatty acid β-oxidation in laying hens

The adverse metabolic impacts of branched-chain amino acids (BCAA) have been elucidated are mediated by isoleucine and valine. Dietary restriction of isoleucine promotes metabolic health and increases lifespan. However, a high protein diet enriched in BCAA is presently the most useful therapeutic strategy for nonalcoholic fatty liver disease (NAFLD), yet, its underlying mechanism remains largely unknown. Fatty liver hemorrhagic syndrome (FLHS), a specialized laying hen NAFLD model, can spontaneously develop fatty liver and hepatic steatosis under a high-energy and high-protein dietary background that the pathogenesis of FLHS is similar to human NAFLD. The mechanism underlying dietary BCAA control of NAFLD development in laying hens remains unclear. Herein, we demonstrate that dietary supplementation with 67 % High BCAA has unique mitigative impacts on NAFLD in laying hens. A High BCAA diet alleviates NAFLD, by inhibiting the tryptophan-ILA-AHR axis and MAPK9-mediated de novo lipogenesis (DNL), promoting ketogenesis and energy metabolism, and activating PPAR-RXR and pexophagy to promote fatty acid β-oxidation. Furthermore, we uncover that High BCAA strongly activates ubiquitin-proteasome autophagy via downregulating UFMylation to trigger MAPK9-mediated DNL, fatty acid elongation and lipid droplet formation-related proteins ubiquitination degradation, activating PPAR-RXR and pexophagy mediated fatty acid β-oxidation and lipolysis. Together, our data highlight moderating intake of high BCAA by inhibiting the AHR/MAPK9 are promising new strategies in NAFLD and FLHS treatment.

Incidence of non-alcoholic fatty liver disease in antiretroviral therapy-naïve people with human immunodeficiency virus who start DTG/ABC/3TC compared to BIC/FTC/TAF at 48-week follow-up.

To determine the incidence of non-alcoholic fatty liver disease (NAFLD) by non-invasive methods in people living with HIV (PLWH). Prospective cohort, in PLWH naïve to antiretroviral therapy, starting bictegravir (BIC) or dolutegravir (DTG) at the Hospital de Infectología "La Raza", in Mexico City, from February 2021 to August 2023. We measured at baseline and 48weeks triglycerides and glucose index (TyG), fatty liver index (FLI), hepatic steatosis index (HSI) and liver ultrasonography; relative risk (RR) for developing NAFLD was determined. At 48weeks, TyG index in BIC-group 4.54 (IQR 4.36-4.75), in DTG-group 4.66 (IQR 4.49-4.80), p = .080; HSI in BIC-group 30.30 (IQR 28.12-33.70), in DTG-group 30.85 (IQR 28.02-34.50), p = .650; FLI in BIC-group 14.88 (IQR 7.91-31.80), in DTG-group 19.49 (IQR 8.49-32.28), p = .729; NAFLD was detected by US in 6 [10.3% (95%CI 4.8%-20.7%)] in BIC-group and, 7 [10.9% (95%CI 6.4%-20.9%)] in DTG-group, p = .916. Risk factors for NAFLD development were baseline BMI ≥25kg/m2, baseline HDL-c <40mg/dL, and FIB-4 >1.3 at 48weeks. There is a high incidence of NAFLD in PLWH who start a second generation INSTI at 48weeks; baseline overweight, low HDL-cholesterol and FIB-4 >1.3 at 48weeks of treatment were independent risk factors for NAFLD development.

Association between phthalates exposure and non-alcoholic fatty liver disease under different diagnostic criteria: a cross-sectional study based on NHANES 2017 to 2018.

Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease. Phthalates have been suggested to influence the development of NAFLD due to their endocrine-disrupting properties, but studies based on nationally representative populations are insufficient, and existing studies seem to have reached conflicting conclusions. Due to changes in legislation, the use of traditional phthalates has gradually decreased, and the phthalates substitutes is getting more attention. This study aims to delve deeper into how the choice of diagnostic approach influences observed correlations and concern about more alternatives of phthalates, thereby offering more precise references for the prevention and treatment of NAFLD. A cohort of 641 participants, sourced from the National Health and Nutrition Examination Survey (NHANES) 2017-2018 database, was evaluated for NAFLD using three diagnostic methods: the Hepatic Steatosis Index (HSI), the US Fatty Liver Indicator (US.FLI), and Vibration Controlled Transient Elastography (VCTE). The urinary metabolite concentrations of Di-2-ethylhexyl phthalate (DEHP), Di-isodecyl phthalate (DIDP), Di-isononyl phthalate (DINP), Di-n-butyl phthalate (DnBP), Di-isobutyl phthalate (DIBP), Di-ethyl phthalate (DEP) and Di-n-octyl phthalate (DnOP) were detected. The association between NAFLD and urinary phthalate metabolites was evaluated through univariate and multivariate logistic regression analyses, considering different concentration gradients of urinary phthalates. Univariate logistic regression analysis found significant correlations between NAFLD and specific urinary phthalate metabolites, such as Mono-(2-ethyl-5-oxohexyl) phthalate (MEOHP), Mono-(2-ethyl-5-hydroxyhexyl) phthalate (MEHHP), Mono-2-ethyl-5-carboxypentyl phthalate (MECPP), and Mono-(carboxyisoctyl) phthalate (MCiOP), across different diagnostic criteria. In a multivariate logistic regression analysis adjusting only for demographic data, MEOHP (OR = 3.26, 95% CI = 1.19-8.94, p = 0.029), MEHHP (OR = 3.98, 95% CI = 1.43-11.1, p = 0.016), MECPP (OR = 3.52, 95% CI = 1.01-12.2, p = 0.049), and MCiOP (OR = 4.55, 95% CI = 1.93-10.7, p = 0.005) were positively related to NAFLD defined by HSI and VCTE. The correlation strength varied with the concentration of phthalates, indicating a potential dose-response relationship. Adjusting for all covariates in multivariate logistic regression, only MCiOP (OR = 4.22, 95% CI = 1.10-16.2, p = 0.044), as an oxidative metabolite of DINP, remained significantly associated with NAFLD under the VCTE criterion, suggesting its potential role as a risk factor for NAFLD. This research highlights a significant association between DINP and NAFLD. These findings underscore the need for further investigation into the role of the phthalates substitutes in the pathogenesis of NAFLD and the importance of considering different diagnostic criteria in research.

Non-Invasive Serum Markers of Non-Alcoholic Fatty Liver Disease Fibrosis: Potential Tools for Detecting Patients with Cardiovascular Disease.

Non-alcoholic fatty liver disease (NAFLD), one of the most common chronic liver diseases with a prevalence of 23%-25% globally, is an independent risk factor for cardiovascular diseases (CVDs). Growing evidence indicates that the development of NAFLD, ranging from non-alcoholic fatty liver (NAFL), non-alcoholic steatohepatitis (NASH), advanced fibrosis to cirrhosis, and even hepatocellular carcinoma, is at substantial risk for CVDs, which clinically contribute to increased cardiovascular morbidity and mortality. Non-invasive serum markers assessing liver fibrosis, such as fibrosis-4 (FIB-4) score, aspartate transaminase-to-platelet ratio index (APRI), and NAFLD fibrosis score (NFS), are expected to be useful tools for clinical management of patients with CVDs. This review aims to provide an overview of the evidence for the relationship between the progression of NAFLD and CVDs and the clinical application of non-invasive markers of liver fibrosis in managing patients with CVDs.

Influence of chronic wartime stress on the course of non-alcoholic fatty liver disease

Objective — to analyze levels of chronic stress hormones in patients with non‑alcoholic fatty liver disease (NAFLD), who were exposed to psychogenic factors of wartime. Materials and methods. Examinations involved 88 patients with NAFLD, form them 38 subject were examined in the pre‑war period (comparison group), 50 patients were under the influence of stress during hostilities (main group). Control group consisted of 20 practically healthy subjects. Patients’ groups matched by age. The main group consisted of 28 men and 22 women, the comparison included 17 men and 21 women. The mean age in the main group was (51.6±7.7) years and in the comparison group (51.8±8.1) року. The trophological status of patients was monitored with the use of Body Composition Monitor.Omron BF­­511; hormones of chronic stress were assessed with solid‑phase enzyme immunoassay. Results. The assessment of levels of the chronic stress biochemical markers in NAFLD patients during the war demonstrated pronounced hyperprolactinemia in comparison with both control group and NAFLD patients examined in the pre‑war period. The dependence of stress hormone levels on the trophic status of patients and gender was revealed. A direct correlation between the prolactin level and the index of insulin resistance (IR), the percentage of visceral adipose tissue (VAT), and the level of triglycerides (TG) was revealed. A weak positive correlation was also observed between the level of brain‑derived neurotrophic factor (BDNF) and IR, VAT, TG, and C‑reactive protein (CRP). The activity of the inflammatory process depended on the level of dihydroepiandrosterone (DHEA), which confirms the direct correlation between the level of this hormone and ALT and CRP. Conclusions. The dependence of stress hormones levels on the trophic status of patients and gender was revealed. The level of DHEA in NAFLD patients with obesity and overweight significantly exceeded the corresponding indicators in the group with normal body mass index (BMI). In the group of overweight women who experienced negative consequences of military actions, the DHEA level significantly exceeded the corresponding indicators in comparison with the pre‑war period. Under the influence of military actions in patients of the main group, the reaction to stress was accompanied by an increase in prolactin levels in men with normal and overweight weight compared to the pre‑war period. In obese women from the main group, the hyperprolactinemia level significantly exceeded the corresponding values in the groups with normal and excessive BMI. The revealed correlation dependence between the stress hormones levels and metabolic factors indicates the influence of chronic stress on the main pathogenetic mechanisms of the development and progression of NAFLD in the conditions of war.

Identification of common signature genes and pathways underlying the pathogenesis association between nonalcoholic fatty liver disease and heart failure.

Non-alcoholic fatty liver disease (NAFLD) and heart failure (HF) are related conditions with an increasing incidence. However, the mechanism underlying their association remains unclear. This study aimed to explore the shared pathogenic mechanisms and common biomarkers of NAFLD and HF through bioinformatics analyses and experimental validation. NAFLD and HF-related transcriptome data were extracted from the Gene Expression Omnibus (GEO) database (GSE126848 and GSE26887). Differential analysis was performed to identify common differentially expressed genes (co-DEGs) between NAFLD and HF. Gene ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and gene set enrichment analysis (GSEA) were conducted to explore the functions and regulatory pathways of co-DEGs. Protein-protein interaction (PPI) network and support vector machine-recursive feature elimination (SVM-RFE) methods were used to screen common key DEGs. The diagnostic value of common key DEGs was assessed by receiver operating characteristic (ROC) curve and validated with external datasets (GSE89632 and GSE57345). Finally, the expression of biomarkers was validated in mouse models. A total of 161 co-DEGs were screened out in NAFLD and HF patients. GO, KEGG, and GSEA analyses indicated that these co-DEGs were mainly enriched in immune-related pathways. PPI network revealed 14 key DEGs, and SVM-RFE model eventually identified two genes (CD163 and CCR1) as common key DEGs for NAFLD and HF. Expression analysis revealed that the expression levels of CD163 and CCR1 were significantly down-regulated in HF and NAFLD patients. ROC curve analysis showed that CD163 and CCR1 had good diagnostic values for HF and NAFLD. Single-gene GSEA suggested that CD163 and CCR1 were mainly engaged in immune responses and inflammation. Experimental validation indicated unbalanced macrophage polarization in HF and NAFLD mouse models, and the expression of CD163 and CCR1 were significantly down-regulated. This study identified M2 polarization impairment characterized by decreased expression of CD163 and CCR1 as a common pathogenic pathway in NAFLD and HF. The downregulation of CD163 and CCR1 may reflect key pathological changes in the development and progression of NAFLD and HF, suggesting their potential as diagnostic and therapeutic targets.

Ablating the glutaredoxin-2 (Glrx2) gene protects male mice against non-alcoholic fatty liver disease (NAFLD) by limiting oxidative distress

In the present study, we investigated the consequences of deleting the glutaredoxin-2 gene (Glrx2−/−) on the development of non-alcoholic fatty liver disease (NAFLD) in male and female C57BL6N mice fed a control (CD) or high-fat diet (HFD). We report that the HFD induced a significant increase in body mass in the wild-type (Wt) and Glrx2−/− male, but not female, mice, which was associated with the hypertrophying of the abdominal fat. Interestingly, while the Wt male mice fed the HFD developed NAFLD, the deletion of the Glrx2 gene mitigated vesicle formation, intrahepatic lipid accumulation, and fibrosis in the males. The protective effect associated with ablating the Glrx2 gene in male mice was due to enhancement of mitochondrial redox buffering capacity. Specifically, liver mitochondria from male Glrx2−/− fed a CD or HFD produced significantly less hydrogen peroxide (mtH2O2), had lower malondialdehyde levels, greater activities for glutathione peroxidase and thioredoxin reductase, and less protein glutathione mixed disulfides (PSSG) when compared to the Wt male mice fed the HFD. These effects correlated with the S-glutathionylation of α-ketoglutarate dehydrogenase (KGDH), a potent mtH2O2 source and key redox sensor in hepatic mitochondria. In comparison to the male mice, both Wt and Glrx2−/− female mice displayed almost complete resistance to HFD-induced body mass increases and the development of NAFLD, which was attributed to the superior redox buffering capacity of the liver mitochondria. Together, our findings show that modulation of mitochondrial S-glutathionylation signaling through Glrx2 augments resistance of male mice towards the development of NAFLD through preservation of mitochondrial redox buffering capacity. Additionally, our findings demonstrate the sex dimorphisms associated with the manifestation of NAFLD is related to the superior redox buffering capacity and modulation of the S-glutathionylome in hepatic mitochondria from female mice.

Impaired SUMOylation of FoxA1 promotes nonalcoholic fatty liver disease through down-regulation of Sirt6

Abnormal SUMOylation is implicated in non-alcoholic fatty liver disease (NAFLD) progression. Forkhead box protein A1 (FoxA1) has been shown to protect liver from steatosis, which was down-regulated in NAFLD. This study elucidated the role of FoxA1 deSUMOylation in NAFLD. NAFLD models were established in high-fat diet (HFD)-induced mice and palmitate acid (PAL)-treated hepatocytes. Hepatic steatosis was evaluated by biochemical and histological methods. Lipid droplet formation was determined by BODIPY and Oil red O staining. Target molecule levels were analyzed by RT-qPCR, Western blotting, and immunohistochemistry staining. SUMOylation of FoxA1 was determined by Ni-NTA pull-down assay and SUMOylation assay Ultra Kit. Protein interaction and ubiquitination were detected by Co-IP. Gene transcription was assessed by ChIP and dual luciferase reporter assays. Liver FoxA1 knockout mice developed severe liver steatosis, which could be ameliorated by sirtuin 6 (Sirt6) overexpression. Nutritional stresses reduced Sumo2/3-mediated FoxA1 SUMOylation at lysine residue K6, which promoted lipid droplet formation by repressing fatty acid β-oxidation. Moreover, Sirt6 was a target gene of FoxA1, and Sirt6 transcription activity was restrained by deSUMOylation of FoxA1 at site K6. Furthermore, nutritional stresses-induced deSUMOylation of FoxA1 promoted the ubiquitination and degradation of FoxA1 with assistance of murine double minute 2 (Mdm2). Finally, activating FoxA1 SUMOylation delayed the progression of NAFLD in mice. DeSUMOylation of FoxA1 at K6 promotes FoxA1 degradation and then inhibits Sirt6 transcription, thereby suppressing fatty acid β-oxidation and facilitating NAFLD development. Our findings suggest that FoxA1 SUMOylation activation might be a promising therapeutic strategy for NAFLD.

Birthweight influences liver structure, function and disease risk: Evidence of a causal association.

Low birthweight is an issue during pregnancy associated with an increased risk of developing liver disease later in life. Previous Mendelian randomisation (MR) studies which explored this issue have not isolated the direct impact of the foetus on birthweight. In the present study, MR was used to assess whether direct foetal effects on birthweight were causally associated with liver structure, function and disease risk independent of intrauterine effects. We extracted single nucleotide polymorphisms (SNPs) from genome-wide association studies (GWAS) about direct foetal-affected birthweight (321 223 cases) to conduct univariable and multivariable MR analyses to explore the relationships between birthweight and 4 liver structure measures, 9 liver function measures and 18 liver diseases. A two-step MR analysis was used to further assess and quantify the mediating effects of the mediators. When isolating direct foetal effects, genetically predicted lower birthweight was associated with a higher risk of non-alcoholic fatty liver disease (NAFLD) (odds ratios [OR], 95% confidence interval [CI]: 1.61, 1.29-2.02, p < 0.001), higher magnetic resonance imaging [MRI] proton density fat fraction (PDFF) and higher serum gamma glutamyltransferase (GGT). Two-step MR identified two candidate mediators that partially mediate the direct foetal effect of lower birthweight on NAFLD, including fasting insulin (proportion mediated: 22.29%) and triglycerides (6.50%). Our MR analysis reveals a direct causal association between lower birthweight and liver MRI PDFF, as well as the development of NAFLD, which persisted even after accounting for the potential influence of maternal factors. In addition, we identified fasting insulin and triglycerides as mediators linking birthweight and hepatic outcomes, providing insights for early clinical interventions.