Establishment and characterization of a metabolic dysfunction-associated steatotic liver disease model in the male Korean field mouse (Apodemus peninsulae): A comparison with the male C57BL/6J mouse.

Nuclear receptor retinoid-related orphan receptor α deficiency exacerbates high-fat diet-induced cardiac dysfunction despite improving metabolic abnormality

Metabolic associated fatty liver disease is a novel concept defined as fatty liver associated with metabolic disorders. We investigated the effect of metabolic associated fatty liver disease on hepatocellular carcinoma patient mortality. A total of 624 patients with hepatocellular carcinoma between 2012 and 2020 were enrolled in this retrospective study. Hepatic steatosis was diagnosed using computed tomography or magnetic resonance imaging. Metabolic associated fatty liver disease was defined based on the proposed criteria in 2020. Propensity score matching was performed for patients with metabolic associated fatty liver disease and those without the condition. A Cox proportional hazards regression model was used to evaluate the association between metabolic associated fatty liver disease and hepatocellular carcinoma patient outcomes. Patients with hepatocellular carcinoma and metabolic associated fatty liver disease tended to achieve better outcomes than did those without metabolic associated fatty liver disease after matching (p<0.001). Metabolic associated fatty liver disease was significantly associated with better prognosis in patients with concurrent hepatitis B infection (p<0.001). Moreover, high levels of hepatitis B viral DNA in serum samples was associated with a significantly increased risk of death in patients without non-metabolic associated fatty liver disease (p=0.045). Additionally, the association between metabolic associated fatty liver disease and survival in hepatitis B virus-related hepatocellular carcinoma was similar in all subgroups based on metabolic traits. Metabolic associated fatty liver disease increases the survival rate of patients with hepatocellular carcinoma and hepatitis B virus infection. The potential interaction of steatosis and virus replication should be considered for future research and clinical treatment strategies.

Increasing evidence has demonstrated that the heat shock protein 70 (HSP70) gene may be closely associated with tissue fibrosis; however, the association between HSP70 and liver fibrosis remains to be fully elucidated. The present study hypothesized that geranylgeranylacetone (GGA) exerts beneficial effects on liver fibrosis though upregulation of the expression of HSP70. Liver fibrosis was induced in rats using carbon tetrachloride (CCl4). The rats were subsequently divided into three groups: Control group, CCl4 model group and CCl4 model + GGA group. Liver fibrosis in the rats was evaluated using hematoxylin and eosin staining, Masson's trichrome staining and Sirius red staining. The levels of serum alanine aminotransferase, aspartate aminotransferase and total bilirubin were determined using an automated biochemistry analyzer. The levels of total hepatic hydroxyproline were also determined. The expression levels of α-smooth muscle actin (α-SMA) and transforming growth factor-β1 (TGF-β1) were determined using immunofluorescence staining and western blotting, and the protein expression levels of HSP70 were determined using western blotting. The CCl4-induced rats exhibited liver fibrosis, increased hydroxyproline content, impaired liver function, upregulated expression levels of the α-SMA and TGF-β1 pro-fibrogenic proteins, and increased expression of HSP70, compared with the control group. These changes were attenuated by treatment with GGA. These results demonstrated that GGA exerted beneficial effects in CCl4-induced liver fibrosis via upregulating the expression of HSP70.

BACKGROUND The relationship between different subgroups of type 2 diabetes (T2D) and the progression of metabolic dysfunction-associated steatotic liver disease (MASLD) and liver fibrosis has not been thoroughly studied. This study aims to determine the association between T2D subgroups and the risk of developing advanced liver fibrosis using the Fibrosis-4 (FIB-4) index, a non-invasive marker for assessing liver fibrosis risk. MATERIAL AND METHODS A total of 1205 patients with T2D were categorized into 4 distinct subgroups: severe insulin-deficient diabetes (SIDD), severe insulin-resistant diabetes (SIRD), mild obesity-related diabetes (MOD), and mild age-related diabetes (MARD). The FIB-4 index was calculated for each patient to estimate the degree of liver fibrosis, with the following cutoff points: <1.3 indicating no or mild fibrosis, 1.3-2.67 suggesting moderate fibrosis, and >2.67 indicating advanced fibrosis (F3-F4). Logistic regression was used to compare the odds of advanced fibrosis across these subgroups. RESULTS The SIRD subgroup exhibited significantly higher odds of advanced liver fibrosis (F3-F4), compared with the other subgroups, as indicated by elevated FIB-4 scores (P<0.05). In contrast, the SIDD and MOD subgroups had lower odds of advanced fibrosis, while the MARD subgroup showed an intermediate association. CONCLUSIONS The findings suggest that the FIB-4 index, as a noninvasive assessment tool, effectively stratifies liver fibrosis risk among different T2D subgroups. This stratification can inform more personalized management strategies for patients with MASLD, underscoring the importance of accounting for the heterogeneity within T2D in clinical assessments of liver fibrosis risk.

Early fibrosis has been described in renal allografts and implicated in the progression of chronic allograft nephropathy (CAN). The precise factors implicated in the initiation and progression of early allograft fibrosis remain uncertain. We studied retrospectively 23 cadaveric renal allograft recipients over a 3-year period, who had paired renal biopsies (Bx) (at implantation and as clinically indicated) within 3 months of transplantation (Tx). Eight of them have progressed over an average period of 3.16 +/- 0.83 years to CAN. Histological evaluation of interstitial fibrosis (IF) relied on point count analysis of Masson's trichrome (MT) staining as well as immunostainable collagens III (iCol III) and IV (iCol IV). The severity of the IF scores was correlated with the clinical, biochemical and histological parameters. The nature and severity of the interstitial inflammatory infiltrate were also evaluated by immunofluorescence. In addition, patients were subdivided into those whose fibrosis progressed (> 50% increase in IF/iCol IlI; Group 1) and non-progressors (< 50% increase in fibrosis score; Group 2) in an attempt to determine discriminatory features. In the whole group, there was a significant increase in the IF score, as estimated by MT staining and iCol III, from implantation to follow-up Bx (p = 0.0027 and p = 0.0088, respectively). The changes in iCol IV were not significant. Further, the increase in interstitial inflammatory infiltrate of total T lymphocytes, and not of macrophages, from implantation (modal category = 2) to follow-up (modal category = 0) was significant (p = 0.0121). The predictive value of such increase was significant (R2 = 0.617, p = 0.03). The donor's age (R2 = 0.892, p = < 0.0001), death from cerebrovascular accident (CVA) (R2 = 0.822, p = 0.047), as well as recipient's body weight (R2 = 0.892, p = 0.001), male gender (R2 = 0.687, p = 0.041) and elevated mean arterial pressure (MAP) (R2 = 0.892, p = < 0.0001) were all significant risk factors for early IF. Delayed graft function (DGF) proved to be a significant predictor of early IF (R2 = 0.822, p = 0.003) and became more significant in the presence of superimposed acute rejection (AR) (p = 0.0001). Proteinuria > 1 g/day (R2 = 0.882, p = 0.004) and hypertriglyceridemia > 2.25 mmol/l (R2 = 0.808, p = < 0.0001) were also associated with early IF. Of the implantation histological parameters, iCol III proved to be a highly significant predictor of early IF (R2 = 0.892, p = < 0.0001). Interestingly, the predictive value of iCol III for graft survival in terms of CAN was significant (Cox p = 0.088). Group 1 progressor patients (n = 10) were all males (p = 0.038) and received their kidneys from donors who died from CVAs in 90% of cases (p = 0.011). They had, compared to non-progressors, a lower cyclosporin A level (p = 0.047), a higher incidence of AR episodes (80% versus 54%), a higher serum creatinine at 10 days post-Tx (p = 0.005), a higher proteinuria (2.07 +/- 3.89 g/l vs 0.96 +/- 0.97 g/l, p = 0.041) and a higher serum triglyceride (2.48 +/- 1.37 mmol/l vs 1.69 +/- 0.81 mmol/l, p = 0.039) level. 8% of Group 1 patients had DGF compared to 30% in Group 2 (p = 0.023). Of note, the modal category of cytotoxic: helper T lymphocytes ratio was greater than 1 in Group 1 (2:1) patients and not in Group 2 (1:1). Implantation histology, and in particular iCol III, is a predictor of early IF in a subgroup of patients with DGF and AR. Additional risk factors include hypertension, proteinuria and hypertriglyceridemia especially in patients receiving kidneys from older donors who died of CVAs.

Objective: To compare Masson's trichrome (MT), Sirius red (SR) and orcein staining in acute hepatitis (AH) and to correlate them with transient elastography (TE), a noninvasive method to assess hepatic fibrosis. Methods: We evaluated liver stiffness by TE in a cohort of 34 consecutive patients and assessed MT-, SR- and orcein-stained biopsies using the METAVIR scoring system and digital image analysis (DIA). Results: MT and SR both showed severe fibrosis (stage III-IV, DIA = 12.7%). Orcein showed absent or mild fibrosis (stage 0-II, DIA = 4.4%; p < 0.05). In 29/34 cases (85%), stiffness values were >12.5 kPa, in keeping with SR/MT but not with orcein results. Conclusions: Even though in AH true elastic fibrosis is typically absent or mild, TE shows elevated stiffness values, in keeping with SR/MT evaluations. If not properly evaluated in the clinical context, these results would lead to an overestimation of fibrosis. Orcein is the only staining able to evidence the absence of true elastic fibrosis, which is a typical feature of AH. This is the first study comparing different staining procedures performed on AH biopsies by DIA versus TE.

Yet more evidence that MAFLD is more than a name change

Danning tablets alleviate high fat diet-induced obesity and fatty liver in mice via modulating SREBP pathway

Mycotoxins co-contamination of agricultural products poses a serious threat to human and animal health, especially hepatic dysfunction. Zearalenone (ZEN), deoxynivalenol (DON), and aflatoxin B1 (AFB1) are three commonly co-occurring mycotoxins. This study is to determine whether lycopene (LYC) can alleviate hepatic toxicity induced by the co-occurrence of ZEN, DON, and AFB1 in mice. Eighty 6-week-old male ICR mice are divided into four groups: CON group (solvent control), LYC group (10 mgkg-1 LYC), Co-M group (10 mgkg-1 ZEN + 1 mgkg-1 DON + 0.5 mgkg-1 AFB1), and LYC+Co-M group (10 mgkg-1 LYC + 10 mgkg-1 ZEN + 1 mgkg-1 DON + 0.5 mgkg-1 AFB1). The results show that LYC can suppress the co-occurrence of mycotoxin-induced mitochondrial swelling and vacuolization accompanied by dysregulation of indices of mitochondrial dynamics (Mitofusin 1 (Mfn1), Mfn2, Optic atrophy 1 (Opa1), Dynamin-related protein 1 (Drp1), Fission 1 (Fis1) at the mRNA level; DRP1 and FIS1 at the protein level). LYC effectively inhibits co-occurrence of mycotoxin-induced activation of Cytochrome P450 2E1, and early fibrosis, as determined by staining with Masson's trichrome and α-SMA protein. LYC successfully attenuates early hepatic fibrosis mainly through antioxidant activities and prevented mitochondrial injury.

Obesity-induced vascular inflammation is an early pathological change for the development of diabetic nephropathy. Studies have previously demonstrated that high fat diet (HFD) treatment decreased cytochrome P450 epoxygenase-mediated epoxyeicosatrienoic acids (EETs) production which in turn triggers vascular dysfunction and renal inflammation. We hypothesize that HFD up-regulates let-7b miRNA to decrease epoxgenase-mediated EETs production triggering vascular dysfunction and renal injury. Injection of let-7b mimetic has let-7b (1 nmol/day i.v) into rats decreased renal and hepatic cyp2c23 and cyp2j epoxygenases expression (P&lt; 0.05). To determine whether high fat diet up-regulates let-7b to decrease EETs production, we use sEH gene (Ephx2) knock-out mice (KO) as a model for high EETs availability as EETs are rapidly hydrolyzed by the soluble epoxide hydrolase (sEH) to less active metabolites. WT and Ephx2 (-/-) mice were fed normal (ND, 14% fat) or HFD (60 % fat) for 3 months. HFD treatment was associated with 2 fold up-regulation of renal let-7b miRNA and this effect was coincided with down-regulation of cyp2c44 and cyp2j epoxygenases and decreased EETs levels. Decreased EETs levels in HFD fed rats was also associated with significant podocyte loss and elevation in podocalyxin excretion (56±4 ng/day) when compared to rats fed ND (33±7 ng/day) and these changes were significantly reduced in Ephx2 (-/-) mice. Furthermore, HFD treatment markedly elevated T helper cells expressing inflammatory IL-17 (CD3+CD4+TH17+) and decreased anti-inflammatory regulatory T cells (Tregs, CD3+CD4+FOXP3+) when compared to ND and these changes were also significantly attenuated in Ephx2 (-/-) mice. In-vitro, treatment of cultured glomerular endothelial cells with palmitate (200 μM) for 48 hours increased let-7b miRNA expression and decreased cyp2J mRNA and the tight junction protein ZO-1 expression levels and these changes were significantly reduced by transfecting cells with let-7b inhibitor (50 nM). Our data suggest that up-regulation of let-7b miRNA decreases epoxygenase-mediated EETs production to trigger vascular dysfunction and inflammation in the kidney of obese mice via shifting T cells polarization to favor inflammatory Th17 cells activation.

The first 1000 days of life are critical for shaping long-term metabolic and organ health. We aimed to evaluate whether sugar restriction during the first 1000 days of life, due to the United Kingdom sugar rationing policy (1942-1953), reduced the risk of metabolic dysfunction-associated steatotic liver disease (MASLD) and related liver outcomes in adulthood. We conducted a quasi-natural experiment using UK Biobank data, comparing individuals exposed to sugar rationing in utero and early childhood with those born after rationing ended. Primary outcomes included severe MASLD, major adverse liver outcomes, cirrhosis, and liver cancer, with secondary outcomes assessed by biochemical and imaging biomarkers. Mediation analysis was performed to examine the role of metabolic syndrome traits. Among 63,698 participants, early-life sugar rationing was associated with a 30% reduced risk of MASLD (hazard ratio, 0.70; 95% confidence interval, 0.58-0.85 for in utero plus 1-2 years exposure), 17% lower risk of metabolic dysfunction-associated steatotic liver disease with alcohol consumption, 35% for major adverse liver outcomes, and 32% for cirrhosis. The protective effect increased with longer exposure. Mediation analysis indicated that metabolic syndrome traits explained over 60% of the protective association. Improved liver biomarkers and imaging findings (magnetic resonance imaging-proton density fat fraction reduced by 0.50% and cT1 by 52.2 ms) in the rationed group supported these results. Findings were consistent across genotypes (PNPLA3, TM6SF2) and alcohol intake (metabolic dysfunction-associated steatotic liver disease with alcohol consumption) subgroups. Early-life sugar restriction during critical developmental windows is associated with lower adult risk of MASLD and liver complications, primarily mediated by improved metabolic profiles. Policy efforts targeting sugar intake in pregnancy and early childhood may provide durable benefits for liver and metabolic health.

Noninvasive tests (NITs), e.g., Fibrosis-4 Index (FIB-4) and vibration-controlled elastography (VCTE), have been used to identify patients with metabolic dysfunction-associated steatotic liver disease (MASLD) at high risks for hepatocellular carcinoma (HCC). This study investigates the cost-effectiveness of NITs to identify MASLD patients with advanced liver fibrosis and initiate HCC surveillance. A cost-utility analysis using a Markov model compared no use of NITs with 3 NIT strategies: (i) FIB-4 and VCTE (FIB-4/VCTE), (ii) FIB-4 alone, and (iii) VCTE alone to identify advanced liver fibrosis and initiate HCC surveillance with biannual ultrasonography with alpha-fetoprotein in 4 MASLD populations: (i) general patients with MASLD, (ii) MASLD patients with body mass index (BMI) > 30 kg/m 2 , (iii) MASLD patients with diabetes, and (iv) MASLD patients with 3 metabolic traits (diabetes, hypertension, and BMI >30). FIB-4/VCTE was the most cost-effective approach across all groups, showing the lowest incremental cost-effectiveness ratio, followed by FIB-4 alone and VCTE alone. In the general MASLD population, both FIB-4/VCTE and FIB-4 alone were cost-effective in the United States, whereas only FIB-4/VCTE was cost-effective in Thailand. For MASLD patients with BMI >30, all strategies were cost-effective in the United States, whereas only FIB-4/VCTE was cost-effective in Thailand. In MASLD patients with diabetes or 3 metabolic traits, all strategies were cost-effective in the United States, whereas FIB-4/VCTE and FIB-4 alone were cost-effective in Thailand. Using FIB-4/VCTE to initiate HCC surveillance is cost-effective for patients with MASLD. If VCTE is unavailable, FIB-4 alone is a cost-effective alternative for MASLD patients with diabetes or 3 metabolic traits.

Early detection of cardiac fibrosis in diabetic mice by targeting myocardiopathy and matrix metalloproteinase 2

From "Burnt-Out" to "Burning-Out": Capturing Liver Fat Loss in Patients With Advanced Metabolic Dysfunction-Associated Steatotic Liver Disease From a Dynamic Perspective.

Noninvasive evaluation of hepatic fibrosis is a growing scientific effort in medicine and is of particular interest as early diagnosis is important for the timely prevention and treatment of liver fibrosis and cirrhosis. Non-invasive markers of liver fibrosis have recently been developed as an alternative to liver biopsy. Aim of the work: The aim of this study was to assess the diagnostic value of serum microRNA 122 in Egyptian chronic hepatitis C virus infected patients. This may be a useful tool as a non invasive diagnostic test to detect early stages of fibrosis in comparison to 4 non invasive indexes (APRI, Forns, FIB-4, and FI) vs. control and their ability to differentiate between mild and moderate fibrosis stages. This study was conducted on 40 chronic hepatitis C patients diagnosed by liver biopsy and PCR and 20 apparently healthy volunteers who served as control group (III). Liver fibrosis was staged according to the METAVIR scoring system and consequently patients were classified in two groups of liver fibrosis: mild fibrosis (I) and moderate fibrosis (II). The mean expression level of microRNA-122 was significantly higher in both patient groups (I and II) as compared to the control group (III) (p < 0.001 for each). While there was no statistically significant difference in serum miR-122 expression level between group I compared to group II, microRNA 122 and 4 non invasive indexes (APRI, Forns, FIB-4, and FI) were statistically significant for prediction of fibrosis. MicroRNA 122 had the best performing ROC curve for prediction of fibrosis followed by APRI, FI, Forns, and FIB-4. The AUROCs ranged from 0.912 for FIB-4 to 1 for microRNA 122. While FIB-4 and Forns were statistically significant in differentiating mild and moderate fibrosis, FIB-4 had a better AUROC than Forns (0.75 and 0.71, respectively). The study concluded that there was increased expression of mcroiRNA-122 in Egyptian chronic hepatitis C virus (CHCV) infected patients. MicroRNA 122 has a strong potential to serve as one of the novel noninvasive markers of early liver fibrosis.