Liver Tissue Levels Research Articles

Rational Design of a Tandem Activatable Fluorescent Probe for Differential Diagnosis and Therapeutic Assessment of Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is a formidable disease, distinguished by its high aggressiveness and dismal outcomes. Although leucine aminopeptidase (LAP) has been widely employed as a biomarker in biological imaging of HCC, it is still susceptible to interference from false-positive signals activated in injured liver tissues. In this study, based on the significant difference of GSH levels in alcohol-damaged liver tissues and tumor tissues, a dual-tandem activatable probe (PCLT) was designed for differential diagnosis and treatment guidance of HCC by near-infrared fluorescence (NIRF) imaging. This probe comprised a dual-locked hemicyanine dye decorated with a tetraethylene glycol chain and dual-recognition unit of glutathione (GSH) and LAP, which could be sequentially cleaved by GSH and LAP to restore its NIRF signal. PCLT excellently discriminated orthotopic HCC from ALI far earlier (7 days) than histological analysis (28 days) and exhibited higher specificity toward early orthotopic HCC than the single-locked probe (PCL). In addition, PCLT is capable of accurately delineating the tumor contour, assisting in surgical resection of HCC tumors under fluorescence visualization, and noninvasively assessing the antitumor effect of HCC chemotherapy during ferroptosis, thereby presenting promising clinical implications for clinical diagnosis and therapy of HCC.

GlutaR: AHigh-Performance Fluorescent Protein-Based Sensor for Spatiotemporal Monitoring of Glutamine Dynamics In Vivo.

Glutamine is the most abundant amino acid in human blood and muscle, and is integral to a wide variety of functions in cancer cells. However, the inability to monitor the subcellular distribution of glutamine in real-time has obscured understanding of glutamine metabolism under physiological and pathological conditions. Here, we report the development of a genetically encoded fluorescent sensor and demonstrate how this GlnBP-cpYFP fusion "GlutaR sensor" undergoes glutamine-induced conformational changes reflected in detectable fluorescence responses. Obtained after iterative screening of approximately 1,600 variants, GlutaR exhibits a ratiometric readout, fast response kinetics, and high responsivity, and we demonstrate its selectivity for monitoring glutamine fluctuations in multiple cell types. Additionally, using digitonin permeabilization of GlutaR HeLa cells, we generated a calibration curve and performedin situtitration to quantify free glutamine concentrations in subcellular compartments (cytosol, nucleus, mitochondria). Subsequently, we applied GlutaR to investigate how chemical and genetic inhibition of GS and GLS differentially alter glutamine levels in subcellular compartments. Finally, we demonstrate GlutaR's ability to monitor dynamic glutamine levels in muscle and liver tissues of diabetic micein vivo.These findings collectively demonstrate GlutaR as a versatile tool for the spatiotemporal characterization of glutamine metabolism in living cells and tissues.

Short-chain chlorinated paraffins induce liver injury in mice through mitochondrial disorders and disruption of cholesterol-bile acid pathway.

Short-chain chlorinated paraffins (SCCPs) are pervasive organic pollutants recognized for their persistence and bio-toxicity. This study investigated the hepatotoxic mechanisms of SCCPs at environmentally relevant concentration (0.7 μg/kg). The results showed that SCCPs exposure in mice resulted in dysregulated blood and liver lipids, marked by elevated cholesterol levels. Additionally, liver function was compromised, as indicated by increased levels of aspartate aminotransferase, alanine aminotransferase, and alkaline phosphatase. Histopathological examination of liver tissue post-SCCPs exposure revealed hepatocyte enlargement, vacuolar degeneration, and mild ballooning degeneration. Mechanistically, SCCPs induced mitochondrial abnormalities, evidenced by heightened Hoechst 33258 fluorescence, and augmented reactive oxygen species and malondialdehyde levels in liver tissue. This was accompanied by a reduction in total antioxidant capacity, culminating in elevated apoptosis markers, including cytochrome C and caspase-3. Moreover, SCCPs perturbed hepatocellular energy metabolism, characterized by increased glycolysis, lactic acid, and fatty acid oxidation, alongside a disruption in the tricarboxylic acid cycle and a decline in mitochondrial energy metabolic function. Furthermore, SCCPs exposure downregulated the expression of genes involved in bile acid synthesis (cyp27a1, fxr, and shp), thereby precipitating the cholesterol-bile acid metabolism disorders and cholesterol accumulation. Collectively, these findings underscore that SCCPs, even at environmentally relevant levels, can induce lipid dysregulation, mitochondrial disorders and cholesterol deposition in the hepatocytes, contributing to liver damage. The study's insights contribute to a comprehension of SCCPs-induced hepatotoxicity and may inform potential preventative and treatment targets for hepatic damage associated with SCCPs exposure.

Conditional over-expression of heme-oxygenase 1 in myelomonocytic cells reduces AngII-induced hypertension and vascular inflammation in mice

Abstract Background Systemic arterial Hypertension is one of the most important risk factor responsible for morbidity and mortality world-wide. Angiotensin II (Ang II), a potent vasoconstrictor and key player in the renin-angiotensin-aldosterone system (RAAS) is responsible for vascular dysfunction, inflammation, and tissue damage. Heme oxygenase-1 (HO-1) overexpression may confer antioxidant and anti-inflammatory properties in Ang II-induced hypertension through its action on heme catabolism, generating carbon monoxide (CO), ferritin and biliverdin/bilirubin by the action of biliverdin reductase A (BLVRA). Methods and results Male 9–12-weeks-old HO-1indxLySMCre/wt and LySMCre/wt mice were infused with AngII (1mgAngII/Day/Kg) for 7 days to induce hypertension and compared to sham treatment. Blood pressure monitoring by tail-cuff method, and endothelium-dependent vasodilator studies via organ chamber system using acetylcholine (ACh) in isolated aortic segments (~4 mm), revealed that overexpression of HO-1 in myeloid cells resulted in decreased systolic blood pressure and improved endothelial function in AngII-infused HO-1indxLySMCre/wt mice compared to controls. Concurrently, increased BLVRA expression in liver tissue and elevated plasma bilirubin levels were detected by transcriptome analysis and high-performance liquid chromatography, respectively. These results were supported by a reduction in the expression of inducible cytokines and cell adhesion molecules (CAMs) in the aorta, assessed using qPCR. Bone marrow transplant experiments demonstrated that bone marrow from LysMcre mice in HO-1indxLySMCre/wt mice abrogated the protective effect of HO-1, resulting in endothelial damage and increased blood pressure, potentially due to decreased liver BLVRA expression and the accumulation of bone marrow-derived cells in the vessel wall in response to AngII. Additionally, adeno-associated viral vector (AAV) transfection targeting specifically BLVRA activity in liver lead to an increase in blood pressure values and worse endothelium relaxation, in parallel with higher oxidative stress and the blood neutrophils concentration, as assessed in whole blood by using oxidative burst method and blood count system respectively. Conclusion The overexpression of HO-1 in myeloid cells confers anti-inflammatory protection in AngII-induced arterial hypertension in mice. Myeloid cells bone marrow-derived overexpressing HO-1 regulate the differentiation and infiltration of pro-inflammatory immune cells in the vasculature. BLVRA expression and bilirubin levels downstream of HO-1 play a critical role in protecting against vascular damage by reducing leukocyte infiltration.

Selenium Attenuates Ethanol-induced Hepatocellular Injury by Regulating Ferroptosis and Apoptosis.

Ferroptosis is a newly identified type of cell death which is strongly linked to the development of several diseases. Whereas, the role of ferroptosis in the improvement of ethanol-induced hepatocytes injury by selenium has not been confirmed. In this study, an in vitro cell damage model was established using half inhibition concentration of ethanol to induce NCTC clone 1469. Cell activity, lipid peroxidation, apoptosis and the expression of markers related to ferroptosis pathway was determined. A mouse model of alcoholic liver disease (ALD) was constructed and the effectiveness of selenium and ferrostatin-1 in treating ALD in vivo was assessed by serum liver function tests, tissue staining and immunohistochemistry for ferroptosis related proteins. Pretreatment with selenomethionine and ebselen significantly improved ethanol-induced reduction in hepatocyte viability, elevated GSH levels and SOD enzyme activity, reduced MDA and iron content, while improving ethanol-induced changes in apoptosis levels and ferroptosis markers GPX4, SLC7A11, and ACSL4, with the effect of Selenomethionine being more significant. In vivo results also indicated that intervention with selenium or ferroptosis inhibitors significantly improved ethanol-induced liver tissue damage, significantly reduced serum ALT and AST levels, upregulated GPX4 and SLC7A11, but reduced ACSL4 protein levels in liver tissue. The process of ethanol damage to hepatocytes is regulated by the ferroptosis pathway. Selenium may exert a beneficial role in ethanol-induced hepatocyte injury by antagonizing oxidative stress and regulating apoptosis and ferroptosis pathways.

Effect of curcumin and cacao extract on interleukin-6 expression on ribociclib-induced hepatotoxicity in rats

Purpose: To assess the relationship between interleukin-6 (IL-6) expression and the protective effect of curcumin and cacao extract on hepatotoxicity caused by ribociclib. Method: Sixty adult male albino rats (9 – 10 weeks old) were utilized in this experiment. The rats were separated into six groups (n = 10 each). Group 1 (control) rats received 2 mL of normal saline daily. Group 2 rats were administered 5 mg/kg ribociclib and group 3 rats received 5 mg/kg ribociclib with a cacao dose of 200 mg/kg daily. Group 4 rats were administered 5 mg/kg ribociclib treated with curcumin (200 mg/kg) daily. Groups 5 and 6 were administered cacao and curcumin doses of 200 mg/kg daily. All rats were sacrificed and liver immunohistochemical and histological examinations were performed. Result: The findings demonstrated that ribociclib administration caused liver damage and morphological changes at a histological level in liver tissue. The immunohistochemical investigation revealed that IL-6 levels were significantly increased in group 2 and control group (p < 0.05). Furthermore, co-administration of curcumin and cacao with ribociclib suppressed the levels of IL-6 expression in the liver tissue of rats compared to control group 1. Conclusion: Co-administration of curcumin and cacao protects the liver against ribociclib-induced liver damage. It is necessary to determine the effect of long-term administration of these extracts on the functions of some organs such as the liver and kidney.

The effect of aqueous extracts and isolation proteinous compound from clove buds on serumglucose, triglyceride , glutathione and malnodialdehyde levels in alloxan induced diabetic mice

This study was included preparing acold aqueous extract of clove buds The study also comprised the isolation and studying the proteinous compound,which was seperated using gel filtration technique and determined approximately molecular weight of this isolated compound(6799) dalton . The aim of the study demonstrate effects of the crude aqueous, non proteinous extract, proteinous precipitate and proteinous compound on serum glucose, total cholesterol, triglyceride and high density lipoprotein-cholesterol levels, also glutathione and malondialdehyde levels in liver and kidney tissues in diabetic mice-induced alloxan.Extracts were administerated interaperitioneally. The results were indicated that the crude aqueous, non proteinous extract, proteinous precipitate and proteinous compound which were caused asignificant decrease in serum glucose, total cholesterol, triglyceride levels and malondialdehyde level in liver and kidney tissues, while aproteinous precipitate don’t show asignificant decrease in serum cholesterol level but all extracts don’t show asignificant changes in serum high density lipoprotein-cholesterol level in alloxan induced diabetic mice, on other wise all extracts showed asignificant increase in glutathione level in liver and kidney tissues except non proteinous extract and proteinous precipitate which were caused asignificant increase in glutathione level in liver tissue only.

The cytoplasmic-nuclear transport of DDX3X promotes immune-mediated liver injury in mice regulated by endoplasmic reticulum stress

Immune-mediated liver injury is a common characteristic of various liver diseases, including autoimmune and viral hepatitis. Here, we investigated the role of DEAD-box helicase 3, X-linked (DDX3X) in immune-mediated liver injury. Liver injury was induced in C57BL/6J mice via concanavalin A (Con A). DDX3X hepatocyte-specific knockout (DDX3XΔHep) mice and control (DDX3Xfl/fl) mice were utilized to investigate the role of DDX3X in liver injury. Primary hepatocytes were treated with tunicamycin (TM) to induce ER stress in vitro. The expression of DDX3X in patients with various liver diseases was evaluated. Hepatic DDX3X expression increased, and DDX3X translocated from the cytoplasm to the nucleus during Con A-induced liver injury. DDX3X deficiency ameliorated mouse liver injury and reduced ER stress in liver tissue. The inhibition of ER stress with 4-PBA significantly attenuated liver injury while decreasing DDX3X levels in liver tissue. However, the upregulation of hepatic DDX3X expression reversed Con A-induced liver injury and negated the protective effect of 4-PBA. Mechanistically, the nuclear translocation of DDX3X promoted ER stress-induced apoptosis through the transcriptional induction of CHOP. Moreover, DDX3X was elevated and translocated into the nucleus in patients with HBV-LF and AIH. Additionally, serum DDX3X levels markedly increased in patients with HBV-LF, and a consistent decrease in DDX3X was associated with a good prognosis. The cytoplasmic-to-nuclear translocation of DDX3X promotes ER stress-induced apoptosis, which is an obligatory step that drives hepatic necrosis and tissue damage. Notably, DDX3X is a potential therapeutic target for immune-mediated liver injury.

Hepatoprotective potentials of Usnea longissima Ach. and Xanthoparmelia somloensis (Gyelnik) Hale extracts in ethanol-induced liver injury

In our study, the antioxidant and anti-inflammatory effects of different lichen applications were investigated in rats using an experimental ethanol toxicity model. 48 rats were used in the study and they were divided into 6 groups with 8 rats in each group. These groups were: control, ethanol (2 g/kg), ethanol + Usnea longissima Ach. (200 mg/kg), ethanol + Usnea longissima Ach. (400 mg/kg), ethanol + Xanthoparmelia somloensis (Gyelnik) Hale (100 mg/kg) and ethanol + Xanthoparmelia somloensis (Gyelnik) Hale (200 mg/kg). The experimental work continued for 21 days. Lichen extracts and ethanol were administered by gavage to rats divided into groups. According to the experimental protocol, the experimental animals were sacrificed and their liver tissues were isolated. Biochemical parameters in serum, histological examinations, oxidative stress and inflammation parameters both at biochemical and molecular level in liver tissues were performed. Oxidative stress and inflammatory response were increased in the liver tissue of rats treated with ethanol for 21 days, and liver functions were impaired. It was found that U. longissima and X. somloensis extracts showed good antioxidant activity and conferred protective effects against ethanol-induced oxidative stress and inflammation. This could be attributed to the presence of secondary metabolites in the extract, which act as natural antioxidants and could be responsible for increasing the defence mechanisms against free radical production induced by ethanol administration.

Evaluation of the Efficacy of Ozone Therapy on Liver Tissue in the Treatment of Sepsis in Rats with Cecal Perforation.

Background and Objectives: Sepsis and its related complications are associated with high morbidity and mortality, often leading to liver damage. Ozone, a molecule with anti-inflammatory and antioxidant properties, may offer protective effects. This study aimed to evaluate the therapeutic and protective impact of ozone on liver injury in a rat model of sepsis induced by cecal ligation and perforation (CLP). Material and Methods: A total of 36 rats were randomly divided into five groups: control (Group C), ozone (Group O), cecal ligation and perforation (Group CLP), ozone + cecal ligation and perforation (Group O+CLP), and cecal ligation and perforation + ozone (Group CLP+O). In the ozone groups, 4 mL of ozone (20 µ/mL) was injected intraperitoneally. Biochemical and histopathological parameters were evaluated in liver tissue samples obtained at the end of 24 h. Results: Polymorphonuclear leukocyte and monocyte infiltration and the total injury score were significantly reduced in the ozone-treated groups compared to the CLP group (p < 0.001). Tumor necrosis factor and interleukin 10 levels in the rat liver tissue were significantly reduced in the O+CLP and CLP+O groups compared to the CLP group, with the O+CLP group showing a more substantial decrease than the CLP+O group (p < 0.001). Thiobarbituric acid reactive substances and glutathione s-transferase levels were significantly lower in the ozone-treated groups compared to the CLP group (p < 0.001). Catalase activity was significantly elevated in the O+CLP group compared to the CLP group (p < 0.001). Serum aspartate transaminase, alanine transaminase, gamma-glutamyl transferase, and total bilirubin were significantly increased in the CLP group and decreased in the ozone-treated groups (p < 0.001, p < 0.001, p = 0.01, p < 0.001 respectively). Conclusions: Administering ozone to rats one hour before the CLP significantly mitigated liver damage, showing a more pronounced effect compared to administering ozone one hour after CLP. The results indicate that ozone could serve a protective function in managing sepsis-induced liver damage.

Genomic and transcriptomic landscape to decipher the genetic basis of hyperpigmentation in Lanping black-boned sheep (Ovis aries)

BackgroundLanping black-boned sheep (LPB) represent a distinctive mammalian species characterized by hyperpigmentation, resulting in black bone and muscle features, in contrast to their conventional counterparts exhibiting red muscle and white bone. The genetic basis underlying LPB hyperpigmentation has remained enigmatic.MethodsIn this study, we conducted whole-genome sequencing of 100 LPB and 50 Lanping normal sheep (LPN), and integrated this data with 421 sequenced datasets from wild and domestic sheep, shedding light on the genetic backdrop and genomic variations associated with LPB. Furthermore, we performed comparative RNA-Seq analysis using liver sample to pinpoint genes implicated in the pigmentation process. We generated a comprehensive dataset comprising 97,944,357 SNPs from 571 sheep, facilitating an in-depth exploration of genetic factors.ResultsPopulation genetic structure analysis revealed that the LPB breed traces its origin back to LPN, having evolved into a distinct breed. The integration of positively selected genes with differentially expressed genes identified two candidates, ERBB4 and ROR1, potentially linked to LPB hyperpigmentation. Comparative analysis of ERBB4 and ROR1 mRNA relative expression levels in liver, spleen, and kidney tissues of LPB, in comparison to Diqing sheep, revealed significant upregulation, except for ERBB4 in the liver. Gene expression heatmaps further underscored marked allelic frequency disparities in different populations.ConclusionOur findings establish the evolutionary lineage of the LPB breed from LPN and underscore the involvement of ERBB4 and ROR1 genes in melanin synthesis. These results enhance our comprehension of the molecular basis of hyperpigmentation and contribute to a more comprehensive depiction of sheep diversity.

Protective Effect of Corilagin on Alcoholic Liver Injury

Objectives The herbal product corilagin is renowned for its liver-protecting benefits and antioxidant properties. However, the protective role of corilagin in alcoholic liver injury remains unexplored. The objective of this study is to delve into the protective effects of corilagin on alcoholic liver injury, both in HepG2 cells and a mouse model. Methods To assess the antioxidant capacity of corilagin in vitro, DPPH and ABTS assays were conducted. HepG2 cells were pretreated with 800 mM ethanol for 24 hours, followed by corilagin treatment for another 24 hours, to evaluate its liver-protecting activity. For in vivo studies, ICR mice, with and without corilagin treatment, were given ethanol for 4 weeks to induce alcoholic liver injury. At the end of the study, serum and liver tissue samples were collected to analyze liver function, enzymatic antioxidants, reactive oxygen species (ROS) levels, alcohol dehydrogenase, and liver tissue histology. Results The findings reveal that corilagin exhibits remarkable free radical scavenging ability and alleviates alcohol-induced cell damage in vitro. In vivo, corilagin significantly reduced liver index and serum levels of aminotransferase aspartate, alanine transaminase, total cholesterol, and triglyceride in mice with alcoholic liver injury. Additionally, corilagin decreased ROS levels in liver tissue and increased the activities of antioxidant enzymes CAT, SOD, and GSH-Px. GSH content increased, while lipid peroxide MDA decreased in liver tissue. Conclusion Our data strongly suggests that corilagin possesses protective effects on alcoholic liver injury, both in vitro and in vivo, which is likely attributed to its exceptional antioxidant capacity. In summary, corilagin holds promise as a potential preventive or therapeutic agent for alcoholic liver injury.

Lipidome and proteome analyses provide insights into Mariana Trench Snailfish (Pseudoliparis swirei) adaptation to the hadal zone

The hadal zone environment, characterized by extreme hydrostatic pressure, low temperatures, and limited food availability, presents substantial survival challenges for deep-sea fish species. In this study, we captured five deep-sea fish species (Bathysaurus mollis, Coryphaenoides rudis, Ilyophis sp., I. brunneus and Pseudoliparis swirei) from the Mariana Trench at depths ranging from 2027 to 7125 ​m, by employing China's “Exploration I and Exploration II”. By combining lipidomic and proteomic analyses, we aimed to explore the genetic basis of adaptive evolution to the hadal zone in fish. The results indicate several key findings: (1) P. swirei (Mariana hadal snailfish, MHS) may enhance energy storage and utilization during prolonged fasting periods by significantly increasing liver tissue levels of cholesterol ester (CE), ether-linked triacylglycerol (TG-O), coenzyme Q (CoQ), and ATPase content; (2) MHS could maintain membrane fluidity under high pressure by increasing the proportion of unsaturated fatty acids while reducing levels of cholesterol and phosphatidylethanolamine (PE) content. (3) The regulation of lipid types and ratios could increase the risk of lipid peroxidation. To counter oxidative stress, MHS likely elevates monounsaturated fatty acid content and enhances antioxidants such as transferrin and heat shock proteins. Overall, this study provides new insights into the adaptive mechanisms of MHS to deep-sea conditions through the lipidome and proteome analyses, thus broadening our understanding of its resilience in the hadal zone.

Improvement of Liver Function (AST and ALT) and Liver Tissue Catalase levels of Wistar rats induced with Alloxan by consuming Catfish oil extract

Background: Background: Type 2 Diabetes Mellitus (T2DM) is characterized by disrupted glucose metabolism, leading to hyperglycemia and insulin resistance, often associated with Secondary Hypertension (SH). Over 80% of SH patients exhibit glucose intolerance, with nearly 30% developing T2DM. There is a strong interaction between T2DM and NAFLD, representing a complex two-way relationship that influences the prognosis of the two diseases. Catfish oil extract boasts unsaturated fatty acids, including DHA, EPA, omega-3, vitamins A, B6, B12, D, and amino acids. Objective: This study aims to assess the impact of Pangasius sp. (catfish) oil extract administration on serum AST and ALT levels, as well as liver tissue MDA and CAT levels in alloxan-induced Wistar rats (Rattus norvegicus). Materials and Methods: Employing a pure experimental approach with a post-test-only control group design, three groups of 9 white male rats (Rattus norvegicus), along with one extra male rat, were included.K1 served as the negative control group (Wistar rats not subjected to any treatment). K2 acted as the positive control group (Wistar rats subjected to alloxan-induced diabetes at 150mg/kg BW). K3 represented the treatment group (diabetic model rats treated with catfish oil extract at 73mg/kg BW). Serum AST and ALT levels, as well as MDA and CAT levels in liver tissue, were measured at the study's conclusion. Results: The highest mean MDA levels in white rat liver tissue were K2 (3034.00 + 525.25 nmol/g), and the lowest was K3(2909.33+351.01nmol/g); the mean CAT liver tissue levels in rats the highest was K1 (1063.42+133.36U/mg), and the lowest was K3(894.78+132.93U/mg), the highest mean serum ALT levels of white rats was K2(230.34+63,58 U/L), and the lowest was K1(151.54+23.12 U/L) and the highest mean serum AST level in rats was K2(448.79+618.90U/L), and the lowest was K1(61.01+14.70U/L). Conclusion: Giving catfish oil supplementation can repair the damage in liver tissue, as evidenced by reductions in MDA levels and serum ALT and AST levels in diabetic model rats within the treatment group. Alloxan induction did not affect liver tissue CAT levels, as evidenced by a consistent decrease in liver tissue CAT levels in both the positive control and treatment groups.

Effect of Polyherbal Formulation on High Fat Diet Induced Obesity in Rats

Obesity is one of the major chronic diseases in which excess body fat accumulates in visceral organs. It is a chronic metabolic disease due to an imbalance between energy intake and expenditure. Various synthetic treatments are available in the market but have side effects such as Joint pain, chest tightness, tiredness, Back and stomach pain. The aim of study was to examine the anti-obesity potential of prepared polyherbal formulation (Hugonia Mystax: Blumea Lacera::1:1) in female Wistar rats. The obesity was induced by a high fat diet. Group 1 Placebo (normal control), Group 2-high fat diet (disease control), Group 3-high fat diet and orlistat (20 mg/kg for last 2 weeks, Group 4 treated with high fat diet and low dose elixir (200 mg/kg i.e. 4ml orally for last 2 weeks of study), Group 5- high fat diet and high dose elixir (400 mg/kg i.e. 8ml orally for last 2 weeks of study). At the end of study rats were subjected to evaluation of physical parameters, liver profile, serum blood glucose level and histopathology of liver and adipose tissue. The result states that formulation has the potential to work on various obesity mechanisms especially on energy intake. The result of the present study concluded that the prepared formulation has anti-obesity potential.

ADH4—a potential prognostic marker for hepatocellular carcinoma with possible immune-related implications

ObjectiveThis study aims to explore ADH4 expression in hepatocellular carcinoma (HCC), its prognostic impact, and its immune correlation to provide novel insights into HCC prognostication and treatment.MethodsHCC prognostic marker genes were rigorously selected using GEO database, Lasso regression, GEPIA, Kaplan-Meier and pROC analyses. The expression of interested markers (ADH4, DNASE1L3, RDH16, LCAT, HGFAC) in HCC and adjacent tissues was assessed by Immunohistochemistry (IHC). We observed that ADH4 exhibited low expression levels in liver cancer tissues and high expression levels in normal liver tissues. However, the remaining four genes did not manifest any statistically significant differences between hepatocellular carcinoma (HCC) tissue and adjacent non-cancerous tissue. Consequently, ADH4 became the primary focus of our research. ADH4 expression was validated by signed-rank tests and unpaired Wilcoxon rank sum tests across pan-cancer and HCC datasets. Clinical significance and associations with clinicopathological variables were determined using Kaplan-Meier, logistic regression and Cox analyses on TCGA data. The ADH4-related immune responses were explored by Spearman correlation analysis using TIMER2 data. CD68, CD4, and CD19 protein levels were confirmed by IHC in HCC and non-cancerous tissues.ResultsADH4 showed significant downregulation in various cancers, particularly in HCC. Moreover, low ADH4 expression was associated with clinicopathological variables and served as an independent prognostic marker for HCC patients. Additionally, ADH4 affects a variety of biochemical functions and may influence cancer development, prognosis, and treatment by binding to immune cells. Furthermore, at the immune level, the low expression pattern of ADH4 is TME-specific, indicating that ADH4 has the potential to be used as a target for cancer immunotherapy.ConclusionThis study highlights the diagnostic, prognostic and immunomodulatory roles of ADH4 in HCC. ADH4 could serve as a valuable biomarker for HCC diagnosis and prognosis, as well as a potential target for immunotherapeutic interventions.

Elucidation of the Role of L-Arginine and Nω-Nitro-L-Arginine in the Treatment of Rats with Different Levels of Hypoxic Tolerance and Exposure to Lead Nitrate

Background/Aims: Individual resistance to hypoxia is an important feature of the physiological profile of an organism, particularly in relation to lead-induced toxicity. Methods: Our study focused on evaluating parameters of mitochondrial oxygen consumption, microsomal oxidation, intensity of lipoperoxidation processes and antioxidant defences in the liver of rats with low (LR) and high (HR) resistance to hypoxia to elucidate the mechanisms of action of L-arginine and the NO synthase inhibitor L-NNA before or after exposure to lead nitrate. Results: Our study suggests that the redistribution of oxygen-dependent processes towards mitochondrial processes under the influence of the nitric oxide precursor amino acid L-arginine is an important mechanism for maintaining mitochondrial respiratory chain function during per os lead nitrate exposure (3.6 mg lead nitrate/kg bw per day for 30 days). Animals were given L-arginine at a dose of 600 mg/kg bw (i.p., 30 min) before and after exposure to lead nitrate or the NO synthase inhibitor Nω-nitro-L-arginine (L-NNA) at a dose of 35 mg/kg bw (i.p., 30 min) before and after exposure to lead nitrate. Our experiments demonstrated the efficacy of using lead nitrate to simulate lead-related toxic processes via Pb levels in liver tissue; we demonstrated significantly reduced levels of nitrites and nitrates, i.e. stable metabolites of the nitric oxide system, in both LR and HR animals. The effect of the amino acid L-arginine stabilised the negative effects of lead nitrate exposure in both groups of LR and HR rats. We observed the efficiency of mitochondrial energy supply processes and showed a greater vulnerability of NADH-dependent oxidation during lead nitrate exposure in the liver of HR rats. Conclusion: L-arginine initiated the processes of oxidation of NADH-dependent substrates in the LR group, whereas in the HR group this directionality of processes was more effective when the role of the nitric oxide system was reduced (use of L-NNA). Our study of key antioxidant enzyme activities in rat liver tissue during lead nitrate exposure revealed changes in the catalase-peroxidase activity ratio. We found different activities of antioxidant enzymes in the liver tissue of rats treated with lead nitrate and L-arginine or L-NNA, with a significant increase in GPx activity in the LR group when L-arginine was administered both before and after exposure to lead nitrate.

Protective Effects of Berberine on Nonalcoholic Fatty Liver Disease in db/db Mice via AMPK/SIRT1 Pathway Activation.

Berberine (BBR) has emerged as a promising therapeutic agent for nonalcoholic fatty liver disease (NAFLD). This study aims to elucidate the underlying molecular mechanisms. In this study, db/db mice were chosen as an animal model for NAFLD. A total of 10 healthy C57BL/6J mice and 30 db/db mice were randomly allocated to one of 4 groups: the normal control (NC) group, the diabetic control (DC) group, the Metformin (MET) therapy group, and the BBR therapy group. The total cholesterol (TC), triacylglycerol (TG), low-density lipoprotein cholesterol (LDL-c), high-density lipoprotein cholesterol (HDL-c), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels in the serum were measured. The glutathione peroxidase (GSH-Px), glutathione (GSH), malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), interleukin (IL)-1β, tumor necrosis factor (TNF)-α and monocyte chemotactic protein 1 (MCP-1) levels in liver tissue were measured. Hematoxylin and eosin (H&E), acid-Schiff (PAS) and TUNEL stanning was performed for histopathological analysis. Western blotting and immunohistochemistry were conducted to detect the expression levels of key proteins in the AMPK/SIRT1 pathway. BBR could improve lipid metabolism, attenuate hepatic steatosis and alleviate liver injury significantly. The excessive oxidative stress, high levels of inflammation and abnormal apoptosis in db/db mice were reversed after BBR intervention. BBR clearly changed the expression of AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1), and their downstream proteins. BBR could reverse NAFLD-related liver injury, likely by activating the AMPK/SIRT1 signaling pathway to inhibit oxidative stress, inflammation and apoptosis in hepatic tissue.

A near infrared fluorescent probe for monitoring peroxynitrite abnormalities in non-alcoholic fatty liver

Peroxynitrite (ONOO−) has emerged as a critical biomarker for detecting pathological alterations in biological stroma. Notably, it has strong correlation with the pathogenesis of conditions like non-alcoholic fatty liver disease (NAFLD), underlining its utility in both identifying and elucidating such diseases. In this work, we report a novel fluorescent probe, TEM-NS, innovatively crafted using an enhanced dicyanoisophorone scaffold, aimed at the precise quantification of ONOO−. Characterized by its turn-on fluorescence response, TEM-NS exhibits an emission profile and a pronounced Stokes shift superior to those of traditional dicyanoisophorone derivatives. Its exceptional specificity and photostability, combined with superior sensitivity in a biological milieu, are noteworthy. Importantly, application of TEM-NS within a NAFLD model substantiated its capability to detect elevated ONOO− levels in liver tissue, thereby affirming its applicability for in-depth analysis of oxidative stress-related disorders.

Using Stable Isotope Techniques to Analyze the Trophic Relationship between Argentine Hake (Merluccius hubbsi) and Anisakidae.

The Argentine hake (Merluccius hubbsi) is a vital fishery species in the Southwest Atlantic, recognized for its substantial economic importance. Previous studies have identified Anisakidae larvae as common parasites of M. hubbsi. However, the nutritional relationships between these parasites and their host remain poorly understood. This study employs stable isotope techniques to investigate the specific nutritional relationships between Anisakidae larvae and different tissues of M. hubbsi. The findings reveal notable differences in δ13C and δ15N compositions between the parasites and their host. The lower δ13C values in parasites compared to host tissues indicate the utilization of different carbon sources. The δ15N values of the parasites partially overlap with those of the host's stomach, indicating that the parasites primarily derive nutrients from the host's stomach. Nutritional niche indicators show that parasites have a broad carbon range (CR) and nitrogen range (NR), suggesting a high diversity in nutritional sources. The trophic discrimination factor (ΔTDF), which represents the difference in stable isotope values between host tissues and parasites, was analyzed for both δ13C and δ15N. The ΔTDFδ13C between the host liver and the parasites showed the greatest variation, indicating a strong dependence of the parasites on the liver's carbon sources. In contrast, variations in ΔTDFδ15N between host tissues and parasites were minimal. Analyzing ΔTDF across different stages of gonadal maturity in the host fish indicates that, as the gonads of the host fish mature, ΔTDFδ13C between host tissues and parasites significantly decreases (p < 0.01). The Kruskal-Wallis test showed significant differences in ΔTDFδ13C values among different parasite infection levels in muscle, liver, and stomach tissues, while no significant differences were found for ΔTDFδ15N values. These findings offer valuable insights into the nutritional relationships between parasites and hosts, aiding in a better understanding of the growth conditions and habitats of M. hubbsi.