Markers Of Liver Injury Research Articles

6-Hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline Demonstrates Anti-Inflammatory Properties and Reduces Oxidative Stress in Acetaminophen-Induced Liver Injury in Rats.

We examined the effects of 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline on markers of liver injury, oxidative status, and the extent of inflammatory and apoptotic processes in rats with acetaminophen-induced liver damage. The administration of acetaminophen caused the accumulation of 8-hydroxy-2-deoxyguanosine and 8-isoprostane in the liver and serum, as well as an increase in biochemiluminescence indicators. Oxidative stress resulted in the activation of pro-inflammatory cytokine and NF-κB factor mRNA synthesis and increased levels of immunoglobulin G, along with higher activities of caspase-3, caspase-8, and caspase-9. The administration of acetaminophen also resulted in the development of oxidative stress, leading to a decrease in the level of reduced glutathione and an imbalance in the function of antioxidant enzymes. This study discovered that 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline reduced oxidative stress by its antioxidant activity, hence reducing the level of pro-inflammatory cytokine and NF-κB mRNA, as well as decreasing the concentration of immunoglobulin G. These changes resulted in a reduction in the activity of caspase-8 and caspase-9, which are involved in the activation of ligand-induced and mitochondrial pathways of apoptosis and inhibited the effector caspase-3. In addition, 6-hydroxy-2,2,4-trimethyl-1,2-dihydroquinoline promoted the normalization of antioxidant system function in animals treated with acetaminophen. As a result, the compound being tested alleviated inflammation and apoptosis by decreasing oxidative stress, which led to improved liver marker indices and ameliorated histopathological alterations.

Rifampicin-induced ER stress and excessive cytoplasmic vacuolization instigate hepatotoxicity via alternate programmed cell death paraptosis in vitro and in vivo

AimsRifampicin-induced hepatotoxicity is a primary cause of drug-induced liver injury (DILI), posing a significant challenge to its continued clinical application. Moreover, the mechanism underlying rifampicin-induced hepatotoxicity remains unclear. Main methodsHuman hepatocyte line-17 (HHL-17) cells were treated with an increasing dose of rifampicin for 24 h, and male Wistar rats were given rifampicin [150 mg/kg body weight (bw)] orally for 28 days. Viability assay, protein expression, and cell death assays were analyzed in vitro. Moreover, liver serum markers, body/organ weight, H&E staining, protein expression, etc., were assayed in vivo. Key findingsRifampicin induced a dose-dependent hepatotoxicity in HHL-17 cells (IC50; 600 μM), and increased the serum levels of liver injury markers, e.g., alanine transaminase (ALT) and aspartate transaminase (AST) in rats. Rifampicin-induced cell death was non-apoptotic and non-necroptotic both in vitro and in vivo. Further, excessive cellular vacuolization and reduced expression of Alix protein confirmed the induction of paraptosis both in vitro and in vivo. In addition, a significant increase in the endoplasmic reticulum (ER) stress markers (e.g., BiP, CHOP, and total polyubiquitinated proteins) was detected, demonstrating the induction of ER stress and altered protein homeostasis. Interestingly, rifampicin-induced hepatotoxicity was associated with the inhibition of autophagy and enhanced reactive oxygen species (ROS) generation in HHL-17 cells. Furthermore, inhibition of protein synthesis by cycloheximide (CHX) suppressed paraptosis by alleviating rifampicin-induced ER stress and ROS generation. SignificanceRifampicin-induced hepatotoxicity involves ER stress-driven paraptosis as a novel mechanism of its toxicity that may be targeted to protect liver cells from rifampicin toxicity.

The Role of Gold Nanorods in Detecting Circulating microRNAs as Biomarkers in Liver Diseases

Liver disease is one of the most significant diseases in humans. Various factors can lead to liver degeneration and necrosis, fatty liver, cirrhosis, etc. This study evaluated the diagnostic value of circulating miRNAs in liver injury and examined the role of gold nanorods (AuNRs) in the detection of biological indicators. We collected data from patients with liver injury in three groups. Healthy volunteers were used as the control group, and the levels of serum markers of liver injury were detected in all participants. The circulating miRNAs associated with liver injury were screened by searching the database, and circulating miRNAs were detected by qRT-PCR. The sensitivity and specificity of circulating miRNAs in diagnosing liver injury were analyzed using receiver operating characteristic curve. AuNRs were prepared to evaluate the concentrations of glucose, cholesterol, uric acid, and alkaline phosphatase (ALP), which were compared with the test results of the kit. We found that the levels of alanine aminotransferase and aspartate aminotransferase changed significantly in patients with drug-induced liver injury (DILI). Total bilirubin levels increased significantly in patients with both non-alcoholic steatohepatitis (NASH) and DILI, but difference between the two groups was insignificant. MDH and adenosine deaminase levels increased in patients with liver injury, whereas paraoxonase-1 levels decreased, which can be combined with alanine aminotransferase levels to diagnose patients with liver injury. Compared with the control group, circulating miR-106-5p, miR-194-5p, and miR-29b-3p were specifically expressed in tissues with liver injury: miR-106-5p expression was lowest in NASH, miR-194-5p expression was highest in chronic hepatitis B (CHB) and NASH, and miR-29b-3p expression was highest in CHB. Receiver operating characteristic curve analysis revealed that miR-106-5p, miR-194-5p, and miR-29b-3p can be used to diagnose patients with NASH, DILI, and CHB, respectively. The combination of miR-194-5p and miR-29b-3p showed good diagnostic ability. The results of biological indicator detection using AuNRs were similar to those detected by the kit. Therefore, AuNRs are a promising candidate for biological detection. This study validated the diagnostic value of circulating miRNAs in liver disease and the application of AuNRs in the detection of biological indicators, providing more theoretical basis for the role of circulating miRNAs in disease prevention and diagnosis.

Potential of circulating receptor-interacting protein kinase 3 levels as a marker of acute liver injury

The pathogenesis of acute liver failure (ALF) involves cell death. Necroptosis is a newly suggested programmed cell death, and receptor-interacting protein kinase 3 (RIPK3) has been reported as a marker for necroptosis. However, there are few reports on necroptosis in ALF. Therefore, we evaluated the role of cell death markers such as cytokeratin (CK) 18, cleaved CK (cCK) 18, and RIPK3 in ALF, as well as cytokines and hepatocyte growth factor (HGF). Seventy-one hospitalized patients with acute liver injury (38 nonsevere hepatitis [non-SH]/22 severe hepatitis [SH]/11 ALF) were studied. No significant difference was found for cytokines, but a substantial increase in HGF levels was found following the severity of hepatitis. The non-SH group had lower levels of CK18 and cCK18 than the SH/ALF group. RIPK3 was significantly lower in the non-SH/SH group than in the ALF group. HGF, RIPK3, and albumin levels were found to be important predictive variables. The present study suggests that cCK18, CK18, and RIPK3 are associated with the severity of hepatitis. RIPK3 and other markers related cell death may be useful for understanding the pathogenesis of ALF and as a prognostic marker of acute liver injury.

Prolonged warm ischemia time increases mitogen-activated protein kinase activity and decreases perfusate cytokine levels in ex vivo rat liver machine perfusion.

Machine perfusion is increasingly being utilized in liver transplantation in lieu of traditional cold static organ preservation. Nevertheless, better understanding of the molecular mechanisms underlying the ischemia-reperfusion injury (IRI) during ex vivo perfusion is necessary to improve the viability of liver grafts after transplantation using machine perfusion technology. Since key cellular signaling pathways involved in hepatic IRI may allow a chance for designing a promising approach to improve the clinical outcomes from this technology, we determined how warm ischemia time (WIT) during procurement affects the activity of mitogen-activated protein kinase (MAPK) and perfusate concentration of cytokines in an ex vivo rat liver machine perfusion model. Male Sprague-Dawley rats underwent in situ hepatic ischemia with varying WIT (0, 10, 20, 30 min, n = 5 each), and subsequently 3 h of cold ischemia time and 2 h of machine perfusion prior to determining the degree of MAPK activation-phosphorylation and cytokine concentration in liver tissue and perfusates, respectively. Our data revealed a strong correlation between incremental WIT and a series of liver injury markers, and that prolonged WIT increases ERK1/2 and p54 JNK phosphorylation during machine perfusion. Notably, specific cytokine levels (MCP-1, MIP-2, GRO/KC, IL-10, and IL-5) were inversely correlated with the phosphorylation levels of ERK1/2, p38 MAPK, and p46/p54 JNK. These results suggest that MAPK activation, specifically ERK1/2 and p54 JNK phosphorylation, have potential as a biomarker for hepatic IRI pathophysiology during machine perfusion. Elucidation of their functional significance may lead to designing a novel strategy to increase the clinical benefit of machine perfusion.

Does ambient air pollution influence biochemical markers of liver injury? Findings of a cross-sectional population-based survey

BackgroundThere is limited evidence supporting an adverse effect of ambient air pollution on the liver. ObjectivesTo test the association between exposure to residential air pollution and serum biochemical indicators of liver injury. MethodsWe used a nationally representative sample of 32,989 participants aged 3–79 years old who participated in the Canadian Health Measures Survey between 2007 and 2019. Cross-sectional associations were assessed by generalized linear mixed models incorporating survey-specific sampling weights. ResultsThe joint effect of an interquartile range (IQR) increase in nitrogen dioxide (NO2), ozone (O3) and fine particulate matter (PM2.5) was positively and significantly associated with all measures of liver injury adjusting for age, sex, education, income, smoking, alcohol consumption, body mass index (BMI), total cholesterol, diabetes, hypertension, and physical activity. The ranking of effect sizes from largest to smallest percent increases were 8.72% (95% confidence interval [CI] 7.56, 9.88) for alanine aminotransferase (ALT), 5.54% (95%CI 3.31, 7.77) for gamma-glutamyl transferase (GGT), 4.81% (95%CI 3.87, 5.74) for aspartate aminotransferase (AST), 2.46% (95%CI 0.26, 4.65) for total bilirubin (TBIL) and 1.18% (95%CI 0.62, 1.75) for alkaline phosphatase (ALP). Findings were not significantly different when stratified by age (≤16, >16 yr), sex, smoking (current, other), cholesterol (≤6.18, >6.18 mmol/l) and BMI (<30, ≥30 kg/m2). DiscussionThese findings suggest that ambient air pollution may have a relatively small impact on the liver, but these changes may have significant impact from a population health perspective, considering the ubiquitous nature of air pollution, or for individuals exposed to very high levels of air pollution.

Gut ecological networks reveal associations between bacteria, exercise, and clinical profile in non-alcoholic fatty liver disease patients.

Gut microbial dysbiosis has been observed in non-alcoholic fatty liver disease (NAFLD). The beneficial impact of exercise, the recommended lifestyle change for NAFLD patients, might be mediated by the gut microbiome (GM). However, the exact taxonomic and functional signatures associated with the host's clinical and biochemical improvement during exercise in NAFLD patients have not been elucidated yet. To investigate the impact of exercise on GM and reveal GM structures associated with NAFLD improvement during exercise. Stool samples from a 12-week randomized controlled exercise study on NAFLD subjects (N = 39) were retrieved, and shotgun metagenomics was performed at baseline and endpoint. Differential correlation network and enrichment analysis were applied to characterize the GM taxonomic and functional changes during the exercise intervention and to associate GM changes with biomarkers of liver status and metabolic dysregulation. Network analysis demonstrated that exercise induced significant changes in the bacterial interactome, which were associated with waist circumference; resting metabolic rate; plasma fasting concentrations of triglyceride, glucose, insulin, and glycated hemoglobin A1c; and homeostasis model assessment for insulin resistance. Grouping the patients in the exercise group (N = 20) as responders (N = 13) and non-responders (N = 7) using their intrahepatic lipid content (IHL) change allowed us to identify bacteria consortia contributing to the levels of alanine fermentation, methanol-, creatinine-, and protocatechuate degradation and as a result to the plasma concentrations of liver injury markers alanine transaminase, gamma-glutamyl transaminase, and aspartate transaminase. We showed that even though exercise has not a significant impact on the alpha- and beta-diversity of NAFLD patients, it leads to a significant re-structuring of the gut bacteria interactome and that specific structural changes can be linked with improvements in IHL. IMPORTANCE Our study is applying a community-based approach to examine the influence of exercise on gut microbiota (GM) and discover GM structures linked with NAFLD improvements during exercise. The majority of microbiome research has focused on finding specific species that may contribute to the development of human diseases. However, we believe that complex diseases, such as NAFLD, would be more efficiently treated using consortia of species, given that bacterial functionality is based not only on its own genetic information but also on the interaction with other microorganisms. Our results revealed that exercise significantly changes the GM interaction and that structural alterations can be linked with improvements in intrahepatic lipid content and metabolic functions. We believe that the identification of these characteristics in the GM enhances the development of exercise treatment for NAFLD and will attract general interest in this field.

Liver injury in cynomolgus monkeys following intravenous and intrathecal scAAV9 gene therapy delivery

Hepatotoxicity associated with intravenous/intrathecal adeno-associated virus (AAV) gene therapy has been observed in preclinical species and patients. In nonhuman primates, hepatotoxicity following self-complementary AAV9 administration varies from asymptomatic transaminase elevation with minimal to mild microscopic changes to symptomatic elevations of liver function and thromboinflammatory markers with microscopic changes consistent with marked hepatocellular necrosis and deteriorating clinical condition. These transient acute liver injury marker elevations occur from 3-4days post intravenous administration to ∼2weeks post intrathecal administration. No transaminase elevation or microscopic changes were observed with intrathecal administration of empty capsids or a "promoterless genome" vector, suggesting that liver injury after cerebrospinal fluid dosing in nonhuman primates is driven by viral transduction and transgene expression. Co-administration of prednisolone after intravenous or intrathecal dosing did not prevent liver enzyme or microscopic changes despite a reduction of T lymphocyte infiltration in liver tissue. Similarly, co-administration ofrituximab/everolimus with intrathecal dosing failed to blockAAV-driven hepatotoxicity. Self-complementary AAV-inducedacute liver injury appears to correlate with high hepatocellular vector load, macrophage activation, and type 1 interferon innate virus-sensing pathway responses. The current work characterizes key aspects pertaining to early AAV-driven hepatotoxicity in cynomolgus macaques, highlighting the usefulness of this nonclinical species in that context.

Liver FoxO1 overexpression is positively associated with the degree of liver injury in cirrhotic patients

Abstract Objectives Chronic liver disease and related complications, cirrhosis and hepatocellular carcinoma, are associated with high mortality. Curative treatments, partial hepatectomy or liver transplantation, have limited applicability in patients with cirrhosis due to the poor liver regenerative capacity. Thus, we need to find new diagnostic and therapeutic alternatives, to block the disease progression and to improve the survival of patients. In this context, preclinical studies have demonstrated the key role of the protein kinase B (Akt) in liver dysfunction, but the status of Akt and its targets in patients with chronic hepatopathy remains unknown. Aims: To determine the activation status of the Akt pathway and their association with liver functionality in cirrhotic patients. Methods This retrospective study includes liver tissue samples from 36 hepatectomized patients with (n=27) and without (n=9) cirrhosis. Multiplex analysis of proteins involved in the Akt/mTOR pathway was performed using a Luminex panel and Western blot. Conventional liver function tests were determined in serum before resection surgery. Results Akt and forkhead box protein O1 (FoxO1) are overexpressed in the liver of cirrhotic patients: (2.1 vs. 1.0 densitometric relative units (DRU); p&lt;0.01, and 9.5 vs. 4.4 DRU; p&lt;0.01, respectively). FoxO1 showed the best correlation with markers of liver injury (aspartate aminotransferase (ASAT): r=0.51, p&lt;0.05; alanine aminotransferase (ALAT): r=0.49, p&lt;0.05), and was the only enzyme in the Akt pathway identified as an independent predictor of ASAT and ALAT levels. Conclusions The intrahepatic expression of FoxO1 could have clinical utility as a potential prognostic marker for patients with advanced liver disease.

Biochemical enzymes effects of analgesics treatment on post operative patients

Albeit 60 years have passed since it opened up on the restorative market, analgesics measurements in patients with liver illness stays a questionable subject. Fulminant hepatic disappointment has been a proven and factual result of analgesics over portion since its presentation, while short and long haul use have both been related with height of liver transaminases, a proxy marker for intense liver injury. From these reports it has been expected that analgesics use ought to be confined or the measurement decreased in patients with persistent liver sickness. We survey the elements that have been suspected to build hazard of hepatocellular injury from analgesics and the pharmacokinetic changes in various pathologies of persistent liver illness which might influence this gamble. We propose that unintentional under-dosing might bring about focuses excessively low to empower viability. Explicit examination to further develop the proof base for endorsing analgesics in patients with various an etiologies of constant liver sickness is required.. Results communicated that the study just with discoveries communicated that the patients which are not related with contamination but rather they impacted disease (498 numbers and 33.2%) for lesser rate and numbers when contrasted and disease related sicknesses (802 Numbers and 53.5%). The various degrees of SGOT results demonstrated after the medications taken pain+ tachyarrthymia (2.5 ± 0.4↑) patients levels of SGOT were fundamentally expanded contrasted with before drugs taken torment embraced patients. The various levels SGPT results made sense of for ensuing to the medications taken torment + aggravation ( 1.65 ± 0.003↓ ) patients levels of SGPT were widely diminished when contrasted with sooner than drugs taken torment. A more sensible manual for help the shriveling youngster expected by clear notification high philological hindrances with social and extreme contrasts. Additionally course expected to help the youngster and other gathering patients gone through chemotherapy radiation for therapy post operatively.

Independent and combined associations of urinary metals exposure with markers of liver injury: Results from the NHANES 2013–2016

BackgroundHeavy metals entering the human body could cause damage to a variety of organs. However, the combined harmful effects of exposure to various metals on liver function are not well understood. The purpose of the study was to investigate the independent and joint relationships between heavy metal exposure and liver function in adults. MethodsThe study involved 3589 adults from the National Health and Nutrition Examination Survey. Concentrations of urinary metals, including arsenic (As), cadmium (Cd), lead (Pb), antimony (Sb), barium (Ba), thallium (Tl), tungsten (W), uranium (U), were determined in urine using inductively coupled plasma mass spectrometry. Data for liver function biomarkers included alanine aminotransferase (ALT), aspartate aminotransferase (AST), gamma-glutamyl transaminase (GGT), and alkaline phosphatase (ALP). Survey-weighted linear regression and quantile g-computation (qgcomp) were employed to evaluate the relationship of urinary metals with the markers of liver injury. ResultsCd, U and Ba were found to have positive correlations with ALT, AST, GGT, and ALP in the survey-weighted linear regression analyses. According to the qgcomp analyses, the total metal mixture was positively correlated with ALT (percent change: 8.15; 95% CI: 3.84, 12.64), AST (percent change: 5.55; 95% CI: 2.39, 8.82), GGT (percent change: 14.30; 95% CI: 7.81, 21.18), and ALP (percent change: 5.59; 95% CI: 2.65, 8.62), and Cd, U, and Ba were the main contributors to the combined effects. Positive joint effects were observed between Cd and U on ALT, AST, GGT and ALP, and U and Ba had positive joint effects on ALT, AST and GGT. ConclusionExposures to Cd, U, and Ba were individually associated with multiple markers of liver injury. Mixed metal exposure might be adversely correlated with markers of liver function. The findings indicated the potential harmful effect of metal exposure on liver function.

Intravascular Ultrasound Guidance for Transjugular Intrahepatic Portosystemic Shunt Creation Reduces Laboratory Markers of Acute Liver Injury

PurposeTo compare the laboratory markers of acute liver injury after transjugular intrahepatic portosystemic shunt (TIPS) creation performed using intravascular ultrasound (IVUS) guidance with those using other techniques. Materials and MethodsThis single-center, retrospective study examined 293 TIPS procedures performed between 2014 and 2022 (160 men; mean age, 57.4 years; 71.7% with ascites, 158 with IVUS). Laboratory changes on postprocedural day (PPD) 1 were classified based on Common Terminology Criteria for Adverse Events (CTCAE) grades and were compared between IVUS and non-IVUS cases. ResultsIVUS cases had a lower baseline Model for End-Stage Liver Disease (MELD) score (12.5 vs 13.7, P = .016), higher pre- (16.8 vs 15.2, P = .009), and post-TIPS (6.6 vs 5.4 mm Hg, P < .001) pressure gradient, smaller stent diameter (9.2 vs 9.9 mm, P < .001), and fewer needle passes (2.4 vs 4.2, P < .001). IVUS predicted a lower PPD 1 CTCAE grade for aspartate transaminase (8.0% vs 22.2% grade ≥ 2, P = .010), alanine transaminase (ALT) (2.2% vs 7.1%, P = .017), and bilirubin (9.4% vs 26.2%, P < .001), findings confirmed using multivariable regression and propensity score analysis. IVUS predicted fewer adverse events (1.3% vs 8.1%, P = .008) and an increased likelihood of PPD 1 discharge (81% vs 59%, P = .004). IVUS was not associated with differences in PPD 30 MELD scores or 30-day survival; however, higher PPD 1 ALT (β = 1.96, P = .008) and bilirubin levels (β = 1.38, P = .004) predicted larger PPD 30 MELD score increase. Higher increases in ALT level predicted worse 30-day survival (hazard ratio, 1.93; P = .021). ConclusionIVUS resulted in less laboratory evidence of acute liver injury immediately following TIPS creation.

Combined GIP receptor and GLP1 receptor agonism attenuates NAFLD in male APOE∗3-Leiden.CETP mice.

Combined glucose-dependent insulinotropic polypeptide receptor (GIPR) and glucagon-like peptide-1 receptor (GLP1R) agonism is superior to single GLP1R agonism with respect to glycemic control and weight loss in obese patients with or without type 2 diabetes. As insulin resistance and obesity are strong risk factors for nonalcoholic fatty liver disease (NAFLD), in the current study we investigated the effects of combined GIPR/GLP1R agonism on NAFLD development. Male APOE∗3-Leiden.CETP mice, a humanized model for diabetic dyslipidemia and NAFLD when fed a high-fat high-cholesterol diet, received subcutaneous injections with either vehicle, a GIPR agonist, a GLP1R agonist, or both agonists combined every other day. GIPR and GLP1R agonism reduced body weight and additively lowered fasting plasma levels of glucose, triglycerides and total cholesterol. Strikingly, we report an additive reduction in hepatic steatosis as evidenced by lower hepatic lipid content and NAFLD scores. Underlying the lipid-lowering effects were a reduced food intake and intestinal lipid absorption and an increased uptake of glucose and triglyceride-derived fatty acids by energy-combusting brown adipose tissue. Combined GIPR/GLP1R agonism also attenuated hepatic inflammation as evidenced by a decreased number of monocyte-derived Kupffer cells and a reduced expression of inflammatory markers. Together, the reduced hepatic steatosis and inflammation coincided with lowered markers of liver injury. We interpretate that GIPR and GLP1R agonism additively attenuate hepatic steatosis, lower hepatic inflammation, ameliorate liver injury, together preventing NAFLD development in humanized APOE∗3-Leiden.CETP mice. We anticipate that combined GIPR/GLP1R agonism is a promising strategy to attenuate NAFLD progression in humans. This work was supported by a grant from the Netherlands CardioVascular Research Initiative: the Dutch Heart Foundation, Dutch Federation of University Medical Centers, the Netherlands Organization for Health Research and Development, and the Royal Netherlands Academy of Sciences [CVON-GENIUS-II] to P.C.N.R., a Lilly Research Award Program [LRAP] Award to P.C.N.R. and S.K., a Dutch Heart Foundation [2017T016] grant to S.K., and an NWO-VENI grant [09150161910073] to M.R.B.; J.F.D.B. is supported by the Nutrition and Health initiative of the University of Groningen; Z.Y. is supported by a full-time PhD scholarship from the China Scholarship Council (201806850094 to Z.Y.).

Normothermic Machine Perfusion of Donor Livers for Transplantation in the United States: A Randomized Controlled Trial.

To compare conventional low-temperature storage of transplant donor livers [static cold storage (SCS)] with storage of the organs at physiological body temperature [normothermic machine perfusion (NMP)]. The high success rate of liver transplantation is constrained by the shortage of transplantable organs (eg, waiting list mortality >20% in many centers). NMP maintains the liver in a functioning state to improve preservation quality and enable testing of the organ before transplantation. This is of greatest potential value with organs from brain-dead donor organs (DBD) with risk factors (age and comorbidities), and those from donors declared dead by cardiovascular criteria (donation after circulatory death). Three hundred eighty-three donor organs were randomized by 15 US liver transplant centers to undergo NMP (n = 192) or SCS (n = 191). Two hundred sixty-six donor livers proceeded to transplantation (NMP: n = 136; SCS: n = 130). The primary endpoint of the study was "early allograft dysfunction" (EAD), a marker of early posttransplant liver injury and function. The difference in the incidence of EAD did not achieve significance, with 20.6% (NMP) versus 23.7% (SCS). Using exploratory, "as-treated" rather than "intent-to-treat," subgroup analyses, there was a greater effect size in donation after circulatory death donor livers (22.8% NMP vs 44.6% SCS) and in organs in the highest risk quartile by donor risk (19.2% NMP vs 33.3% SCS). The incidence of acute cardiovascular decompensation at organ reperfusion, "postreperfusion syndrome," as a secondary outcome was reduced in the NMP arm (5.9% vs 14.6%). NMP did not lower EAD, perhaps related to the inclusion of lower-risk liver donors, as higher-risk donor livers seemed to benefit more. The technology is safe in standard organ recovery and seems to have the greatest benefit for marginal donors.

255-LB: Liver Pyruvate Carboxylase Deletion in Mice Prevents Fibrosis from a NASH Diet

Obesity and T2D are closely associated with Nonalcoholic Fatty Liver Disease, which can advance to Nonalcoholic Steatohepatitis (NASH) with fibrosis. We previously found that liver pyruvate carboxylase (PC) is required for hepatic gluconeogenesis and the concentration of metabolites essential to mediating redox state and antioxidant function. Liver PC knockout (LPCKO) mice are protected from HFD-induced hyperinsulinemia and glucose intolerance despite retaining excess liver fat and exhibiting reduced antioxidant capacity. Here, we report that despite impaired antioxidant markers and increased inflammatory markers on a HFD, PC loss protected the liver from fibrosis and other indices of liver damage induced by a long-term NASH diet. LPCKO and control mice were fed a high fat, high cholesterol Western Diet (WD) for 13-14 months. After 10 mos. of WD, markers of insulin resistance were assessed by lactate/pyruvate, glucose, and insulin tolerance tests. After sacrifice, liver damage was assessed by serum markers and histology. Oxidative stress and inflammation were evaluated by gene expression, MDA concentration, GSH/GSSG and NADPH/NADP+ redox states, and H2O2 emission potential in isolated mitochondria. LPCKO-WD mice were more sensitive to an insulin tolerance test and exhibited lower fasting blood glucose and better glycemic control during a lactate/pyruvate or glucose tolerance test than controls. Liver malate/pyruvate indicated substantially lower NADPH/NADP+ in LPCKO-WD mice, but there were no differences in MDA levels, GSH/GSSG, and H2O2 emission potential. Despite similar hepatic steatosis, inflammation, and ballooning between LPCKO-WD and controls, fibrosis and serum markers of liver injury were markedly lower in LPCKO-WD mice. The data suggest that liver PC deletion improves glycemia and insulin resistance during chronic Western Diet exposure by reducing hepatic gluconeogenesis and prevents fibrosis independent of steatosis, inflammation, ballooning, and oxidative stress. Disclosure M. R. Iñigo-Vollmer: None. B. Kucejova: None. M. N. Mizerska: None. S. Khan: None. P. A. Szweda: None. H. Zaki: None. L. I. Szweda: None. X. Fu: None. S. C. Burgess: None.

Persistent organic pollutants associate with liver disease in a Finnish general population sample.

Persistent organic pollutants (POPs) have multiple adverse effects on human health. Recent studies show a possible association with liver disease, but population-based data are scarce. In this population-based study, we studied the associations between POPs and biomarkers of liver disease and incident liver disease. This study consisted of 2789 adults that participated in the environmental toxin subset of the Finnish health-examination survey, FINRISK 2007. Toxins were measured from serum samples, and standard liver tests and dynamic aspartate aminotransferase-alanine aminotransferase ratio (dAAR) were measured as biomarkers of liver function. Associations between POPs and the biomarkers were then analysed using linear regression. Associations between POPs and incident liver disease (n = 36) were analysed by Cox regression. Organochlorine pesticides (OCPs), polychlorinated biphenyls (PCBs) and several perfluorinated alkyl substances exhibited statistically significant positive associations with several biomarkers of liver injury (betacoefficient per SD 0.04-0.14, p < 0.05). These associations were stronger in subgroups of individuals with obesity or non-alcoholic fatty liver disease. OCPs, PCBs and perfluoro-octanoic acid also had significant positive associations with dAAR, which can be used to predict risk of incident severe liver outcomes (beta coefficient per SD 0.05-0.08, p < 0.05). OCPs and PCBs were also significantly and positively associated with incident liver disease (hazard ratio per SD 1.82 95% CI 1.21-2.73, p < 0.01 and hazard ratio per SD 1.69, 95% CI 1.07-2.68, p < 0.05 respectively). Several POPs show positive associations with markers of liver injury and incident liver disease, suggesting that environmental toxins are important risk factors for chronic liver disease.

An Updated Systematic Review and Meta-Analysis of the Association between the De Ritis Ratio and Disease Severity and Mortality in Patients with COVID-19.

Patients with Coronavirus disease 2019 (COVID-19) often have elevations in markers of liver injury, particularly serum aspartate transaminase (AST) and alanine transaminase (ALT). Such alterations may affect the AST/ALT ratio (De Ritis ratio) and, potentially, clinical outcomes. We conducted an updated systematic review and meta-analysis of the association between the De Ritis ratio and COVID-19 severity and mortality in hospitalized patients. PubMed, Web of Science, and Scopus were searched between 1 December 2019 and 15 February 2023. The Joanna Briggs Institute Critical Appraisal Checklist and the Grading of Recommendations, Assessment, Development, and Evaluation were used to assess the risk of bias and the certainty of the evidence, respectively. Twenty-four studies were identified. The De Ritis ratio on admission was significantly higher in patients with severe disease and non-survivors vs. patients with non-severe disease and survivors (15 studies, weighted mean difference = 0.36, 95% CI 0.24 to 0.49, p < 0.001). The De Ritis ratio was also associated with severe disease and/or mortality using odds ratios (1.83, 95% CI 1.40 to 2.39, p ˂ 0.001; nine studies). Similar results were observed using hazard ratios (2.36, 95% CI 1.17 to 4.79, p = 0.017; five studies). In six studies, the pooled area under the receiver operating characteristic curve was 0.677 (95% CI 0.612 to 0.743). In our systematic review and meta-analysis, higher De Ritis ratios were significantly associated with severe disease and mortality in COVID-19 patients. Therefore, the De Ritis ratio can be useful for early risk stratification and management in this patient group (PROSPERO registration number: CRD42023406916).

CerS6 triggered by high glucose activating the TLR4/IKKβ pathway regulates ferroptosis of LO2 cells through mitochondrial oxidative stress

Lipid metabolism disorders and mitochondrial dysfunction contribute to the progression of diabetes and chronic liver disease (CLD). Ferroptosis, as a form of cell death centered on reactive oxygen species (ROS) accumulation and lipid peroxidation, is closely related to mitochondrial dysfunction. However, whether there exists mechanistic links between these processes remains unknown. Here, to explore the molecular mechanism of diabetes complicated with CLD, we showed that high glucose could restrain the activity of antioxidant enzymes, promote mitochondrial ROS (mtROS) production, and induce a state of oxidative stress in the mitochondria of human normal liver (LO2) cells. We demonstrated that high glucose induced ferroptosis and promoted the development of CLD, which was reversed by the ferroptosis inhibitor Ferrostatin-1 (Fer-1). In addition, the mitochondria-targeting antioxidant Mito-TEMPO was used to intervene LO2 cells in high-glucose culture, and ferroptosis was found to be inhibited, whereas markers of liver injury and fibrosis improved. Furthermore, high glucose could promote ceramide synthetase 6 (CerS6) synthesis through the TLR4/IKKβ pathway. The knockout of CerS6 in LO2 cells showed that mitochondrial oxidative stress was attenuated, ferroptosis was inhibited, and markers of liver injury and fibrosis were ameliorated. In contrast, the overexpression of CerS6 in LO2 cells showed the opposite changes and these changes were inhibited by Mito-TEMPO. In short, we positioned the study of lipid metabolism to a specific enzyme CerS6, with a high degree of specificity. Our findings revealed the mechanism by which the mitochondria act as a bridge linking CerS6 and ferroptosis, confirming that under high glucose conditions, CerS6 promotes ferroptosis through mitochondrial oxidative stress, eventually leading to CLD.

Exosome-derived miR-142-5p from liver stem cells improves the progression of liver fibrosis by regulating macrophage polarization through CTSB.

This study aims to explore the effect of liver stem cells (LSCs)-derived exosomes and the miR-142a-5p carried by them on the process of fibrosis by regulating macrophages polarization. In this study, CCL4 was used to establish liver fibrosis model. The morphology and purity of exosomes (EVs) were verified by transmission electron microscopy, western blotting (WB) and nanoparticle tracing analysis (NTA). Real-time quantitative PCR (qRT-PCR), WB and enzyme-linked immunoadsorption (ELISA) were used to detect liver fibrosis markers, macrophage polarization markers and liver injury markers. Histopathological assays were used to verify the liver injury morphology in different groups. The cell co-culture model and liver fibrosis model were constructed to verify the expression of miR-142a-5p and ctsb. Immunofluorescence of LSCs markers CK-18, epithelial cell adhesion molecule (EpCam), and AFP showed that these markers were up-regulated in LSCs. In addition, we evaluated the ability of LSCs to excrete EVs by labeling LSCs-EVs with PKH67. We found that CCL4 and EVs were simultaneously treated at 50 and 100 μg doses, and both doses of EVs could reduce the degree of liver fibrosis in mice. We tested markers of M1 or M2 macrophage polarization and found that EVs reduced M1 marker expression and promoted M2 marker expression. Further, ELISA was used to detect the secreted factors related to M1 and M2 in tissue lysates, which also verified the above views. Further analysis showed that the expression of miR-142a-5p increased significantly with the increase of EVs treatment concentration and time. Further, in vitro and in vivo LSCs-EVs regulate macrophage polarization through miR-142a-5p/ctsb pathway and affect the process of liver fibrosis. Our data suggest that EVs-derived miR-142-5p from LSCs improves the progression of liver fibrosis by regulating macrophage polarization through ctsb.

Thermoneutral housing does not accelerate metabolic dysfunction-associated fatty liver disease in male or female C57Bl/6J mice fed a Western diet.

Metabolic dysfunction-associated fatty liver disease (MAFLD) represents a growing cause of mortality and morbidity and encompasses a spectrum of liver pathologies. Although dozens of preclinical models have been developed to recapitulate stages of MAFLD, few achieve fibrosis using an experimental design that mimics human pathogenesis. We sought to clarify whether the combination of thermoneutral (TN) housing and consumption of a classical Western diet (WD) would accelerate the onset and progression of MAFLD. Male and female C57Bl/6J mice were fed a nutrient-matched low-fat control or Western diet (WD) for 16 wk. Mice were housed with littermates at either standard temperature (TS; 22°C) or thermoneutral-like conditions (TN; ∼29°C). Male, but not female, mice housed at TN and fed a WD were significantly heavier than TS-housed control animals. WD-fed mice housed under TN conditions had lower levels of circulating glucose compared with TS mice; however, there were select but minimal differences in other circulating markers. Although WD-fed TN males had higher liver enzyme and higher liver triglyceride levels, no differences in markers of liver injury or hepatic lipid accumulation were observed in females. Housing temperature had little effect on histopathological scoring of MAFLD progression in males; however, although female mice retained a level of protection, WD-TN conditions trended toward a worsened hepatic phenotype, which was associated with higher macrophage transcript expression and content. Our results indicate that interventions coupling TN housing and WD-induced MAFLD should be longer than 16 wk to accelerate hepatic steatosis and increase inflammation in both sexes of mice.NEW & NOTEWORTHY Mouse models leading to accelerated fatty liver onset are a useful translational tool. Here we show that coupling thermoneutral-like housing and Western diet feeding in mice for 16 wk does not lead to significant disease progression in either sex, though the molecular phenotype indicates priming of immune-related and fibrotic pathways.