Hepatocyte Hypertrophy Research Articles

Artificial intelligence driven image analysis identifies phenobarbital induced hepatocyte hypertrophy in liver microtissues across species

In vivo treatment of rats with phenobarbital (PB) induces liver enzyme induction associated with hepatocyte hypertrophy. In the present study we used a novel microTMA technology coupled with artificial intelligence (AI) driven image analysis and proteomics analysis to test the hypothesis that PB treatment of rat and human liver microtissues could recapitulate hepatocyte hypertrophy in vitro. Human and rat liver microtissues were treated with PB over a range of concentrations (500 uM - 2000 uM). Fixed liver microtissues were embedded in paraffin in a microTMA mold, sectioned and stained on parallel microTMA sections with H&E and cell type specific markers, respectively. An AI algorithm was trained to identify and measure changes in hepatocyte cytoplasmic area on images of H&E stained microtissue sections. Image analysis with this algorithm showed that treatment of human and rat liver microtissues with PB (500 uM) for 96 h caused significant increases (p ≤ 0.08- p < 0.01) in hepatocyte cytoplasmic area, a hallmark of hypertrophy. Proteomics analysis of control and PB treated liver microtissue samples confirmed this treatment also caused phase1 and phase 2 enzyme induction in both human and rat samples. In conclusion AI driven image analysis of H&E stained liver microtissue FFPE sections shows that this model can recapitulate a PB-induced hypertrophy response.

Protective Role of Sulforaphane against Physiological Toxicity of Triphenyltin in Common Carp (Cyprinus carpio haematopterus).

This experiment mainly explored the protective role of sulforaphane (SFN) against physiological toxicity of triphenyltin (TPT) in Cyprinus carpio haematopterus. In total, 320 Fish (56.90 ± 0.40 g) were randomly divided into four groups with four replicates each. The control group was fed the basal diet, the TPT group (TPT) was exposed to 10 ng L-1 TPT on the basis of the control group, the SFN + TPT group (TPT + SFN) was fed a diet supplemented with 10 mg kg-1 SFN on the TPT group, and the SFN group (SFN) was fed a diet supplemented with 10 mg kg-1 SFN. After 56 days of breeding trials, the results showed that TPT exposure resulted in a remarkable decrease (p < 0.05) in final weight, weight gain rate (WGR), specific growth rate (SGR), and condition factor (CF), but an increase (p < 0.05) in feed conversion ratio (FCR) and hepatosomatic index (HSI) of fish. TPT treatment decreased (p < 0.05) the amounts of hematocrit (Hct) and hemoglobin (Hb), plasma complement component 3 (C3) and C4 contents, alternative complement pathway (ACH50), acid phosphatase (ACP) and lysozyme (LZM) activities, liver glutathione (GSH) content, catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPX) activities, interleukin 10 (IL-10), and SOD mRNA expressions, but increased (p < 0.05) plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities, liver malonaldehyde (MDA) content, tumor Cyclooxygenase 2 (COX2), and necrosis factor α (TNFα), IL-1β, and MDA mRNA expressions. A histological analysis of the liver showed that a higher occurrence rates of the hepatocyte hypertrophy, nuclear disappearance and hepatocyte vacuolization was observed in the hepatocytes of fish exposed to TPT, and it was accompanied by the dilation of hepatic sinusoids. In addition, the toxicity induced by TPT was significantly improved in the groups that were treated with SFN, and SFN was able to improve growth performance and immunity, alleviate TPT-induced changes in inflammatory factors, ameliorate oxidative stress, and increase the activity of antioxidant enzymes (p < 0.05). The addition of SFN also alleviated liver damage caused by TPT and protected the structural integrity of the liver. Overall, these findings suggest that TPT inhibited the growth, immunity, and antioxidant capacity of Cyprinus carpio haematopteru. Dietary SFN could be beneficial for growth promotion, immunity, antioxidant capacity, and protection of liver structural integrity. Therefore, SFN is a prospective feed supplement for ameliorating the damage caused to fish by TPT contamination.

Nodular regenerative hyperplasia: The role of the CK7 immunohistochemistry pattern of expression in diagnosis.

Nodular regenerative hyperplasia (NRH) is a rare vascular disorder of the liver. Clinically, patients present with portal hypertension with or without a cholestatic pattern of injury. Histologically, the liver parenchyma is composed of small nodules of hypertrophic hepatocytes surrounded by atrophic hepatocytes without significant fibrosis. Nodular regenerative hyperplasia is a difficult diagnosis on biopsy specimens, but biopsy remains the gold standard for diagnosis. In this retrospective review, cytokeratin 7 (CK7) immunohistochemistry (IHC) was used to aid in the diagnosis and further characterization of NRH and NRH-like changes. The H&E-stained slides, reticulin, and CK IHC were reviewed for 22 cases. The percentage of hepatocytes staining for CK7 (0%-100%), the location of staining (centrilobular hepatic progenitor cells vs periportal/bile ductular reaction), and the pattern of staining distribution (patchy or diffuse) were recorded for comparison. Of the 22 cases, 9 were CK7 positive. Cases of NRH, however, expressed various degrees of CK7 positivity in centrilobular hepatic progenitor cells, unlike NRH-like changes, which were either CK7 negative or CK7 positive in periportal hepatocytes or in areas of bile ductular reaction. In cases with the appropriate clinical history and histology, CK7 immunohistochemistrycan be performed to distinguish nodular regenerativehyperplasia (primary) and NRH-like changes (secondary). In difficult cases, CK7 positivity in centrilobular hepatic progenitor cells can help confirm the diagnosis of NRH.These data support NRH as a true entity with a distinct pathophysiology from NRH-like changes.

The Short-term Exposure Effect of Chlorpyrifos (20% EC) on Haematological, Biochemical and Histopathological Response of Striped Catfish Pangasianodon hypophthalmus

An experiment of acute toxicity was conducted to find out the lethal concentration 50 (LC50) of chloropyrifos (CPF) for Pangasianodon hypophthalmus. Our findings indicated that the 96-h LC50 value of CPF for P. hypophthalmus was 0.106 mg L-1. Effects of different concentrations of CPF (0.09, 0.1, 0.11, 0.12, and 0.13 mg L-1) for 96-h on hemato-biochemical, enzyme and histological effects were evaluated in this study. Acute exposure induced significant differences in enzymes acid phosphatase, alkaline phosphatase, aspartate aminotransferase, alanine aminotransferase and acetylcholine of blood serum of the treated group as compared to control group. However, significant decreases in serum total protein, globulin, albumin, and triglycerides levels while serum glucose enhanced with consequent increasing doses of CPF exposure. Also, CPF induced significant declines in RBC count, haemoglobin and hematocrit level whereas the exposure groups showed significant rises in WBC and blood indices levels as compared to control group. Furthermore, CPF exposure causes gill curling, epithelium lifting, necrosis, and lamellae degeneration. Hemosiderin, sinusoid dilations, necrosis, malanomacrophage, hepatocyte hypertrophy were observed in liver histology. Reduction of lumen tubule, hyperplasia, increased bowman space and degenerations were seen in kidney. Overall findings indicates that the acute CPF exposure to P. hypophthalmus resulted in significant adverse changes in biochemical, hematological, and histopathological response.

Occurrence of Microplastics in the Tissues of Nile Tilapia (Orechromis niloticus) from Zobe Dam, Katsina State, Nigeria

A study of the detection of microplastics on Oreochromis niloticus in Zobe Reservoir, Katsina State, Nigeria was carried out. Three fish sampling sites were used; the upstream (Tsawatsawa), midstream (Makera), and downstream (Garhi) were selected based on variation of anthropogenic activities. A total number of one hundred and forty-four (144) fish samples with a total weight (MTW) and total length (MTL) varied between 134.1 ±1.50 to 154.2± 3.41g and 17.5 ± 0.10– 22.1± 0.54 cm respectively, were collected and analysed for the presence of microplastics using standard protocols. Fourier Transform Infrared Spectrophotometer (FTIR) was used for the chemical characterization of microplastic composition examined in the fish species. The examination of microplastic occurrence in different sampled fish gills, liver, and kidney revealed the presence of Polystyrene, Polypropylene, Polyethylene terephthalate, Nylon, Polyester, and Polyvinyl chloride. Fish from all sampling sites revealed varying histopathological alterations ranging from lifting of epithelium thickening, distortion of the primary lamellae, and degeneration of secondary lamellae, blood congestion, and massive lamellae degeneration in the gill tissues. Liver tissues had hepatocyte hypertrophy, sinusoidal dilatation and cytoplasmic vacuolation, presence of lipid granules, cellular infiltration, cytoplasmic vacuolation, and haemorrhage. The kidney tissues were presented with Bowman’s capsules hyperplasia, tubular elongation, renal tubular epithelium shrinkage, hyperplasia, and cytoplasmic vacuolization of renal cells. The severity of the damage varies from site to site indicating that the water body is experiencing pollution at varying locations. Our findings provide important insights into the prevalence and distribution of microplastics in this particular environment.

The analysis of the histomorphometric characteristics and ultrastructural features of the liver in response to environmental cycle desynchronization in the wild desert rodent (Gerbillus tarabuli)

The architecture of the hepatic tissue in the nocturnal wild desert rodent (Gerbillus tarabuli) subjected to a shifted LD (Light/Dark) cycle was investigated using transmission electron microscopy and histomorphometry. Investigation involved two groups: the control group (n = 5), exposed to the regular LD cycle (LD: 12:12), and the shifted group (n = 5), exposed to the LD regimen with a prolonged light phase of 8 h, in a day by day alternation with regular LD cycle (LD: 12:12 / LD: 20:4) during 12 weeks. Histological examination showed in the shifted group disorganization of hepatocyte trabeculae, sinusoidal dilatation, and inflammatory cell infiltration around the portal tracts. Morphometric results revealed hypertrophy of hepatocytes with a size of 29.83 ± 1.69 μm vs. 17.52 ± 0.81 μm and of their nuclei diameter with 8.69 ± 0.25 μm vs. 6.55 ± 0.15 μm. This increase is highly significant (p&lt;0.001) and represents an increase of 70.17% and 32.67%, respectively. We also noticed a significant increase in the number of binucleated cells compared to the control group with an average of 9.73 ± 0.98 vs. 5.53 ± 0.38 binucleated hepatocytes per area unit, respectively. Ultrastructural analysis corroborated the histological findings, confirming the installation of inflammatory foci, characterized by portal and intraparenchymal infiltration of leukocytes, proliferation of Kupffer cells and stellate cells, as well as an expansion of collagen fiber deposits. Electron microscopy also revealed in the shifted group some hepatocytic lesional features represented by vacuolar or liquefied cytoplasm associated with degenerative mitochondria and a rarefaction of organelles in addition to an abundance of lysosomes. Additionally, strong glycogen depletion was observed at the metabolic level. This study showed that 12 weeks of lengthening the light phase of the LD cycle, corresponding to the gerbil's resting period, had a discernible impact on hepatocellular activity, making G. tarabuli an interesting model to study the influence of artificial light at night as a circadian rhythm disruptor.

Morphological and Histopathological Liver Abnormalities Caused by Carbamazepine-Induced Injury in Female Albino Mice

Background: The adverse effects of drugs can damage various organs, especially the liver, leading to a hepatic injury known as hepatotoxicity. Drug-induced liver injury (DILI) is challenging nowadays because of the large number of different drugs used, one of the offending medications that cause DILI is carbamazepine (CBZ), since the liver has an array of functions including detoxification, it will deal with several damages caused by exposure to the drugs. Objective: investigate the effect of (CBZ) 20mg/kg/day on female mice liver after 14 and 30 days of treatment on morphological and histopathological levels. Materials and methods: 20mg/kg/day of CBZ was administered orally for (14) days to (10) female mice, another (10) mice were taking the same concentration for 30 days, and control groups were administered tap water. Results: The findings showed that CBZ can cause liver enlargement, changes in liver appearance, distortion in Glisson’s capsule, cytologic alterations, hepatocyte hypertrophy, ballooning degeneration, pyknosis, karyolysis, karyomegaly, sinusoids dilation, increase in the number and sizes of Kupffer cells, fibrosis, glycogen depletion, and cirrhosis. Conclusion: These findings have shown that carbamazepine (CBZ) can cause hepatotoxicity that can manifest into morphological and histopathological changes.

Mode of action analysis for fluxapyroxad-induced rat liver tumour formation: evidence for activation of the constitutive androstane receptor and assessment of human relevance

The fungicide fluxapyroxad (BAS 700 F) has been shown to significantly increase the incidence of liver tumours in male Wistar rats at dietary levels of 1500 and 3000 ppm and in female rats at a dietary level of 3000 ppm via a non-genotoxic mechanism. In order to elucidate the mode of action (MOA) for fluxapyroxad-induced rat liver tumour formation a series of in vivo and in vitro investigative studies were undertaken. The treatment of male and female Wistar rats with diets containing 0 (control), 50, 250, 1500 and 3000 ppm fluxapyroxad for 1, 3, 7 and 14 days resulted in a dose-dependent increases in relative weight at 1500 and 3000 ppm from day 3 onwards in both sexes, with an increase in relative liver weight being also observed in male rats given 250 ppm fluxapyroxad for 14 days. Examination of liver sections revealed a centrilobular hepatocyte hypertrophy in some fluxapyroxad treated male and female rats. Hepatocyte replicative DNA synthesis (RDS) was significantly increased in male rats given 1500 and 3000 ppm fluxapyroxad for 3 and 7 days and in female rats given 50–3000 ppm fluxapyroxad for 7 days and 250–3000 ppm fluxapyroxad for 3 and 14 days; the maximal increases in RDS in both sexes being observed after 7 days treatment. The treatment of male and female Wistar rats with 250–3000 ppm fluxapyroxad for 14 days resulted in significant increases in hepatic microsomal total cytochrome P450 (CYP) content and CYP2B subfamily-dependent enzyme activities. Male Wistar rat hepatocytes were treated with control medium and medium containing 1–100 μM fluxapyroxad or 500 μM sodium phenobarbital (NaPB) for 4 days. Treatment with fluxapyroxad and NaPB increased CYP2B and CYP3A enzyme activities and mRNA levels but had little effect on markers of CYP1A and CYP4A subfamily enzymes and of the peroxisomal fatty acid β-oxidation cycle. Hepatocyte RDS was significantly increased by treatment with fluxapyroxad, NaPB and 25 ng/ml epidermal growth factor (EGF). The treatment of hepatocytes from two male human donors with 1–100 μM fluxapyroxad or 500 μM NaPB for 4 days resulted in some increases in CYP2B and CYP3A enzyme activities and CYP mRNA levels but had no effect on hepatocyte RDS, whereas treatment with EGF resulted in significant increase in RDS in both human hepatocyte preparations. Hepatocytes from male Sprague-Dawley wild type (WT) and constitutive androstane receptor (CAR) knockout (CAR KO) rats were treated with control medium and medium containing 1–16 μM fluxapyroxad or 500 μM NaPB for 4 days. While both fluxapyroxad and NaPB increased CYP2B enzyme activities and mRNA levels in WT hepatocytes, only minor effects were observed in CAR KO rat hepatocytes. Treatment with both fluxapyroxad and NaPB only increased RDS in WT and not in CAR KO rat hepatocytes, whereas treatment with EGF increased RDS in both WT and CAR KO rat hepatocytes. In conclusion, a series of in vivo and in vitro investigative studies have demonstrated that fluxapyroxad is a CAR activator in rat liver, with similar properties to the prototypical CAR activator phenobarbital. A robust MOA for fluxapyroxad-induced rat liver tumour formation has been established. Based on the lack of effect of fluxapyroxad on RDS in human hepatocytes, it is considered that the MOA for fluxapyroxad-induced liver tumour formation is qualitatively not plausible for humans.

The reversal of PXR or PPARα activation-induced hepatomegaly

The activation of pregnane X receptor (PXR) or peroxisome proliferator-activated receptor α (PPARα) can induce liver enlargement. Recently, we reported that PXR or PPARα activation-induced hepatomegaly depends on yes-associated protein (YAP) signaling and is characterized by hepatocyte hypertrophy around the central vein area and hepatocyte proliferation around the portal vein area. However, it remains unclear whether PXR or PPARα activation-induced hepatomegaly can be reversed after the withdrawal of their agonists. In this study, we investigated the regression of enlarged liver to normal size following the withdrawal of PCN or WY-14643 (typical agonists of mouse PXR or PPARα) in C57BL/6 mice. The immunohistochemistry analysis of CTNNB1 and KI67 showed a reversal of hepatocyte size and a decrease in hepatocyte proliferation after the withdrawal of agonists. In details, the expression of PXR or PPARα downstream proteins (CYP3A11, CYP2B10, ACOX1, and CYP4A) and the expression of proliferation-related proteins (CCNA1, CCND1, and PCNA) returned to the normal levels. Furthermore, YAP and its downstream proteins (CTGF, CYR61, and ANKRD1) also restored to the normal states, which was consistent with the change in liver size. These findings demonstrate the reversibility of PXR or PPARα activation-induced hepatomegaly and provide new data for the safety of PXR and PPARα as drug targets.

Histopathology and micronuclei induction as pollution biomarkers in common carp, Cyprinus carpio from southern Iraq

This study aimed to assess the histopathological alterations and micronuclei induction in the gill, kidney and liver of common carp, Cyprinus carpio from Al-Huwaizah Marshes, Al-Hammar Marshes and Al-Sweap River. Thirty C. carpio were sampled at each locality using monofilament gillnets. Most prominent histopathological alterations recorded in the gill included formation venous sinus in primary lamellae, oedema, curling and distortion of primary lamellae fusion of secondary lamellae, hyperplasia of epithelial cells, necrosis, curling of secondary lamellae, severe dilation of lamellar capillaries, clavate lamellae formation at the tip of secondary lamellae, a proliferation of cartilage tissue in primary lamellae and lifting of the lamellar epithelium whereas congestion, fibrosis with proliferative inflammation, necrosis area, degenerated cells, vacuolar degeneration, vacuolation, nuclear pyknosis, hypertrophy of hepatocytes were reported in the liver. In the kidney, the most pronounced changes were necrosis of the hematopoietic tissue, hyaline droplet and activation of melanomacrophage centres. A higher prevalence of alterations was observed in the population from Al-Huwaizah Marshes, however, Al-Hammar population exhibited severe pathologies. Micronuclei frequency was significantly higher at Al-Huwaizah Marsh (p &lt; 0.05) with low frequency being recorded at Al-Sweap River. This study attests to the effectiveness of histopathology and micronucleus induction as potential tools for discriminating the health of populations from different water bodies.

In vitro and in vivo investigation of a thyroid hormone system-specific interaction with triazoles

Alterations in thyroid hormones (TH) and thyroid-stimulating hormone levels are frequently found following exposure to chemicals of concern. Dysregulation of TH levels can severely perturb physiological growth, metabolism, differentiation, homeostasis in the adult and developmental processes in utero. A frequently identified mode of action for this interaction is the induction of hepatic detoxification mechanisms (e.g. SULTs and UGTs), which lead to TH conjugation and elimination and therefore interfere with hormonal homeostasis, fulfilling the endocrine disruptors (EDs) definition. A short-term study in rats with dietary exposure to cyproconazole, epoxiconazole and prochloraz was conducted and hepatocyte hypertrophy, hepatic UGT activity and Phase 1/2 gene expression inductions were observed together with changes in TH levels and thyroid follicular hypertrophy and hyperplasia. To test for specific interaction with the thyroid hormone system, in vitro assays were conducted covering thyroidal I-uptake (NIS), TH transmembranal transport via MCT8 and thyroid peroxidase (TPO) function. Assays for iodothyronine deiodinases (DIO1–DIO3) and iodotyrosine deiodinase (DEHAL1) were included, and from the animal experiment, Dio1 and Dehal1 activities were measured in kidney and liver as relevant local indicators and endpoints. The fungicides did not affect any TH-specific KEs, in vitro and in vivo, thereby suggesting hepatic conjugation as the dominant MoA.

Histopathological Effects of Cadmium Chloride (CdCl2) Exposure on the Liver of Freshwater Fish Heteropneustes fossilis

The histopathological effects of CdCl2 on the liver of H. fossilis by acute and chronic exposure of 25 mg/l, 50 mg/l, and 75 mg/l during interval of 30 and 60 days were studied. Similarly, control groups were operated simultaneously with experimental set-up to study alterations in the morphology and architecture of liver in order to understand the injury. The Liver of H. fossilis Normally, the liver of H. fossilis displayed a bilobed structure consisting of polygonal hepatocytes that were radially disposed around the hepatic veins, which maintained its characteristic architecture with an evident nucleus and interspersed binding connective tissue. Histopathological changes by exposure to CdCl2 Both acute and chronic exposure were associated with hematogenesis characterized by darker fragile livers with hepatocyte hypertrophy. Hepatocytes lost the polygonal shape and were dispersed; cytoplasmic vacuolization, pyknotic changes, and bile canaliculi dilation. Prolonged exposure worsened these effects, and resulted in marked atrophy accompanied by necrosis, cellular shrinkage and tissue distortion. In addition, higher concentrations of CdCl2resulted in enlarged Kupffer cells, ruptured nuclei, cellular necrosis and focal necrosis in liver tissue. Our results suggest that progressive histopathological alterations in the liver of H. fossilis due to CdCl2exposure, signifying the susceptibility of this organ to heavy metal toxicity and its ecological prospects in aquatic environs.

Tryptophan Prevents the Development of Non-Alcoholic Fatty Liver Disease.

The main aim of this research is to study the protective effects of tryptophan on the histomorphological and biochemical abnormalities in the liver caused by a high-calorie diet (HCD), as well as its ability to normalize mitochondrial functions in order to prevent the development of non-alcoholic fatty liver disease (NAFLD). The study was conducted in male Wistar rats aged 3 months at the start of the experiment. Control animals (group I) were fed a standard diet. Group II experimental animals were fed a diet with an excess of fat (45%) and carbohydrates (31%) for 12 weeks. Group III experimental animals also received L-tryptophan at a dose of 80 mg/kg body weight in addition to the HCD. The presence of NAFLD, functional activity, physiological regeneration, and the state of the liver parenchyma and connective tissue were assessed using physiological, morphological, histo-morphometric, biochemical, and biophysical research methods. HCD induced the development of NAFLD, which is characterized by an increase in liver weight, hypertrophy of hepatocytes and an increase in the concentration of lipids, cholesterol and triglycerides in liver tissue. Increased alanine aminotransferase activity in the liver of obese rats also confirm hepatocytes damage. Tryptophan added to the diet lowered the severity of NAFLD by reducing fat accumulation and violations of bioelectric properties, and prevented a decrease in mitochondrial ATP synthesis. The addition of tryptophan can have a potential positive effect on the liver, reducing the severity of structural, biochemical, mitochondrial and bioelectric damage caused by HCD.

Effects of Gardenia Ternifolia Leaf Extract on antioxidant and hepatic enzymes in sniper-induced toxicity in Albino rats

Aim: This study evaluates the effects of Gardenia ternifolia leaf extract on antioxidant and hepatic enzymes in sniper-induced toxicity in albino rats. Methodology: A total of 42 male albino rats weighing 120g to 150g, grouped into 6 groups of 7rats each. Group 1 (Negative control), Group 2 (Positive control), Group 3 (Therapeutic Low Dose), Group 4 (Therapeutic High Dose), Group 5 (Prophylactic Low Dose) and Group 6 (Prophylactic High Dose). After 4 weeks of treatment, the rats were anaesthetized, sacrificed and blood samples collected. The Liver was also harvested for histological analysis. Superoxide dismutase (SOD) and glutathione peroxidase (GPx) were estimated using a rat-specific sandwich-enzyme linked immunosorbent assay (ELISA) method. Aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were carried out using Reitman Frankel method. Alkaline phosphatase (ALP) was carried out using colorimetric phenolphthalein method. The liver tissue was prepared and stained using standard Haematoxylin and Eosin (H &amp; E) staining technique and captured using scope TekTM device. Results: The mean SOD and GPx values for the therapeutic and prophylactic groups were significantly higher (P &lt;.05) when compared with the positive control. The mean SOD and GPx values in the therapeutic groups were not significantly different (P &gt;.05) from the negative control. The mean ALT, AST and ALP values for the therapeutic and prophylactic groups were significantly lower (P &lt;.05) than the positive control but significantly higher (P &lt;.05) than the negative control. Histology analysis of liver indicated normal histoarchitecture in the negative control while in the positive control, there were severe cell degeneration, distortion, necrosis and death of the hepatocytes. The Therapeutic Low Dose and Prophylactic Low Dose groups indicated severe degeneration and hypertrophy of hepatocytes. The Therapeutic High Dose and Prophylactic High Dose groups showed less deposit, well delineated radiating hepatocytes and near normal cytoarchitecture. Conclusion: Administration of Sniper® (dichlorvos) led to an increase in oxidative stress; depletion of antioxidant enzyme levels. It elevated liver enzymes levels, caused histopathological changes and death of cells in the liver. The treatment with leaf extract of Gardenia ternifolia ameliorated the effect of dichlorvos toxicity, restored the anti-oxidant and liver enzyme levels especially the high dose extract.

P53-Mediated Mitochondrial Translocation of EI24 Triggered by ER Stress Plays an Important Role in Arsenic-Induced Liver Damage via Activating Mitochondrial Apoptotic Pathway.

Chronic arsenic poisoning is a public health problem worldwide. In addition to skin lesions, the detrimental effect of arsenic poisoning on liver damage is one of the major issues. Our previous studies demonstrated that endoplasmic reticulum (ER) stress and p53 were associated with arsenic-induced liver damage. Literature has shown that EI24 is involved in hepatocyte hypertrophy; however, the underlying role and mechanism in arsenic-induced liver damage have not been fully elucidated. In this study, we explored the role of ER stress, p53, and EI24 as well as the regulatory relationship in arsenic poisoning populations and L-02 cells treated with distinct concentration NaAsO2 (2.5, 5, 10, and 20μM). Results showed that as with arsenic dose increment, expression levels of ER stress key proteins GRP78, ATF4, and CHOP were significantly enhanced. Additionally, p53 expression in nucleus, p53 phosphorylation at Ser15 and Ser1392, and p53 acetylation at lys382 were significantly increased in NaAsO2-treated L-02 cells. ER stress inhibitor 4-phenylbutyric acid (4-PBA) decreased the expression of p53 phosphorylation at Ser 392, p53 acetylation at lys382, and p53 expression in nucleus. Additionally, in 5μM NaAsO2 condition, p53 inhibitor pifithrin-α (PFT-α) aggravated 5μM NaAsO2-induced GRP78, ATF4, and CHOP expressions, cell apoptosis, and protein-SH consumption. But in 20μM NaAsO2 condition, PFT-α attenuated NaAsO2-induced cell apoptosis. Further results showed that 20μM NaAsO2 facilitated translocation of EI24 from ER to mitochondrion and interaction with VDAC2, leading to activate mitochondrial apoptotic pathway, but not observed in the 5-μM NaAsO2 group. Moreover, PFT-α and 4-PBA inhibited 20μM NaAsO2-induced EI24 expression in mitochondrion. Collectively, our results indicated that arsenic induced p53 activation via ER stress, under relatively low NaAsO2 concentration, NaAsO2-triggered p53 activation protected cells from apoptosis by alleviating ER stress. Another finding was that under relatively high NaAsO2 concentration, NaAsO2-activated p53 facilitated EI24 mitochondrial translocation and caused mitochondrial permeability increase, which represented a switch of p53 from a benefit role to pro-apoptosis function in NaAsO2-treated cells. The study contributed to in-depth understanding the mechanism of arsenic-induced liver damage and providing potential clues for following study.

Hepatic melanomacrophage centers in the arctic cultured fish Cyclopterus lumpus are not indicative of its health state

The lumpfish, Cyclopterus lumpus, holds significant promise as a candidate for large-scale aquaculture production, particularly in its role as a cleaner fish used to manage sea lice infestations in Atlantic salmon Salmo salar farming. Melanomacrophage centers (MMCs) represent polymorphic structures present in the hemolymphopoietic organs of various vertebrates, serving as a widely applicable histological indicator of the fish immune and health status. This study aims to investigate the histochemical characteristics of MMCs within lumpfish livers and to compare MMC density between hatchery-produced (farmed) and wild individuals. Liver samples were collected from 34 lumpfish and subjected to a range of staining techniques, including haematoxylin-eosin, Azan-Mallory's trichrome, Masson-Fontana, Perls-Van Geison, Mallory's hemofuscin, immunohistochemical detection of cytochrome P450 monooxygenase 1 A (CYP1A), and the terminal deoxynucleotidyl transferase-mediated d'UTP nick-end labelling (TUNEL) method. Hepatocytes from hatchery-produced males exhibited notably high lipid content. Additionally, cells showing positive staining with Masson-Fontana, likely associated with the monocyte/macrophage lineage, were identified. Furthermore, small MMCs containing melanin, lipofuscin-ceroids, and ferric ions were detected. While the density of single monocytes/macrophages was markedly higher in hatchery-produced males, no significant discrepancies in MMCs density were observed between wild and hatchery-produced fish, or between males and females of the same origin. The study also revealed the presence of necrotic foci, characterized by hypertrophic hepatocytes positive for both TUNEL and CYP1A staining. These hypertrophic hepatocytes displayed large lipid droplets and pycnotic nuclei, with hatchery-produced males showing a higher numerical density of such foci. In contrast to findings in other fish species, the study found that MMCs did not appear to serve as reliable markers of health status in lumpfish. This conclusion was reached as MMCs density did not exhibit a correlation with necrotic foci or hepatocyte lipid content.

Three-dimensional analysis of ductular reactions and their correlation with liver regeneration and fibrosis.

The liver has multiple regeneration modes, including hepatocellular hypertrophy and self-renewal of hepatocytes. When hepatocyte proliferation is impaired, hepatic progenitor cells may proliferate through ductular reaction (DR), differentiate into hepatocytes, and contribute to fibrosis. However, the three-dimensional spatial relationship between DR and regenerating hepatocytes and dynamic changes in DR associated with fibrosis remain poorly understood. Here, we performed three-dimensional (3D) imaging of cleared 42 liver explants with chronic and acute liver diseases and 4 normal livers to visualize DR. In chronic hepatic liver diseases, such as viral hepatitis, steatohepatitis, autoimmune hepatitis, and cryptogenic cirrhosis, the total length and number of branches of DR showed a significant positive correlation. We studied the spatial relationship between DR and GS-expressing cells using glutamine synthetase (GS) and cytokeratin 19 (CK19) as markers of liver regeneration and DR, respectively. The percentage of CK19-positive cells that co-expressed GS was less than 10% in chronic liver diseases. In contrast, nearly one-third of CK19-positive cells co-expressed GS in acute liver diseases, and chronic cholestatic liver diseases, including primary biliary cholangitis and primary sclerosing cholangitis, showed no co-expression. We also found that DR was longer and had more branching in livers with progressive fibrosis compared to those with regressive fibrosis. Our results suggest that DR displays varying degrees of spatial complexity and contribution to liver regeneration. DR may serve as hepatobiliary junctions that maintain continuity between hepatocytes and bile ducts rather than hepatocyte regeneration in chronic liver diseases.

Histologic changes in the liver and kidney tissues of rats with acute alcohol injury and metabolic correction

Annotation. The liver and kidneys are among the organs that often suffer from the toxic effects of ethanol. The issue of drug correction of alcoholic organ damage, and in particular the role of H2S, remains insufficiently studied. The aim of the study was to evaluate the role of quercetin, hydrogen sulphide donor NaHS and their combination in the correction of morphological changes in the liver and kidneys of rats with acute alcohol injury (AAI). The study was conducted on 25 white male rats weighing 120-130 g, divided into five groups (5 rats in each group). Animals of groups 2-5 were modelled with AAI by intragastric administration of 40% ethanol at a dose of 20 ml/kg for 7 days. In order to correct the AAI, animals of group 3 were administered quercetin (100 mg/kg, intraperitoneally, once/day, for 7 days), group 4 – the hydrogen sulphide donor NaHS-H2O (3 mg/kg, intraperitoneally, once/day, for 7 days), group 5 – both quercetin and NaHS-H2O in the above doses. Animals of group 1 (control) received an equivalent amount of solvents. Histological examinations were performed according to conventional methods. It has been established that in the liver of rats with AAI there are disturbances in the lobular structure, radial ordering of the hepatic beams, signs of inflammation, hypertrophy and fatty infiltration of hepatocytes. In the kidneys, signs of renal glomerular fragmentation, vascular glomerular compaction, tubular epithelial dystrophy, and inflammation are found. Some improvements in the histological structure of the liver and kidneys were observed with quercetin, but they were inferior to those observed with NaHS. Combined therapy with quercetin and NaHS was most effective in restoring the normal structure of the liver and kidneys of rats: the radial arrangement of hepatic beams was restored, the nuclear cytoplasmic index increased, signs of fatty degeneration of hepatocytes and renal epithelial dystrophy decreased, and the activity of the inflammatory response in the organs decreased. The obtained results histologically confirm the feasibility of using hydrogen sulphide donors to enhance the hepatoprotective and nephroprotective effects of quercetin in the setting of AAI.

Histological features of non-alcoholic fatty liver disease revealed in response to mixed vehicle emission exposure and consumption of a high-fat diet in wildtype C57Bl/6 male mice

Non-alcoholic fatty liver disease (NAFLD) is currently plaguing the population at pandemic proportions and is expected to become more prevalent over the next decade. Recent epidemiological studies have demonstrated a correlation between the manifestation of NAFLD and ambient air pollution levels, which is exacerbated by other risk factors, such as diabetes, dyslipidemia, obesity, and hypertension. Exposure to airborne particulate matter has also been associated with inflammation, hepatic lipid accumulation, oxidative stress, fibrosis, and hepatocyte injury. While prolonged consumption of a high-fat (HF) diet is associated with NAFLD, little is known regarding the effects of inhaled traffic-generated air pollution, a ubiquitous environmental pollutant, on the pathogenesis of NAFLD. Therefore, we investigated the hypothesis that exposure to a mixture of gasoline and diesel engine emissions (MVE), coupled with the concurrent consumption of a HF diet, promotes the development of a NAFLD phenotype within the liver. Three-month-old male C57Bl/6 mice were placed on either a low-fat or HF diet and exposed via whole-body inhalation to either filtered (FA) air or MVE (30 µg PM/m3 gasoline engine emissions + 70 µg PM/m3 diesel engine emissions) 6 hr/day for 30 days. Histology revealed mild microvesicular steatosis and hepatocyte hypertrophy in response to MVE exposure alone, compared to FA controls, yielding a classification of "borderline NASH" under the criteria of the modified NAFLD active score (NAS) system. As anticipated, animals on a HF diet exhibited moderate steatosis; however, we also observed inflammatory infiltrates, hepatocyte hypertrophy, and increased lipid accumulation, with the combined effect of HF diet and MVE exposure. Our results indicate that inhalation exposure to traffic-generated air pollution initiates hepatocyte injury and further exacerbates lipid accumulation and hepatocyte injury induced by the consumption of a HF diet, thereby contributing to the progression of NAFLD-related pathologies.

Peroxisome proliferator-activated receptor α agonist induces mouse hepatomegaly through the spatial hepatocyte enlargement and proliferation.

Peroxisome proliferator-activated receptor alpha (PPARα) activation-induced hepatomegaly is accompanied by hepatocyte hypertrophy around the central vein (CV) area and hepatocyte proliferation around the portal vein (PV) area. However, the molecular mechanisms underlying this spatial change of hepatocytes remains unclear. In this study, we examined the characteristics and possible reasons for the zonation distinction of hypertrophy and proliferation during PPARα activation-induced mouse liver enlargement. Mice were injected with corn oil or a typical mouse PPARα agonist WY-14643 (100 mg·kg-1·d-1, i.p.) for 1, 2, 3, 5 or 10 days. At each time point, the mice were sacrificed after the final dose, and liver tissues and serum were harvested for analysis. We showed that PPARα activation induced zonal changes in hepatocyte hypertrophy and proliferation in the mice. In order to determine the zonal expression of proteins related to hepatocyte hypertrophy and proliferation in PPARα-induced liver enlargement, we performed digitonin liver perfusion to separately destroy the hepatocytes around the CV or PV areas, and found that PPARα activation-induced increase magnitude of its downstream targets such as cytochrome P450 (CYP) 4 A and acyl-coenzyme A oxidase 1 (ACOX1) levels around the CV area were higher compared with those around the PV area. Upregulation of proliferation-related proteins such as cell nuclear antigen (PCNA) and cyclin A1 (CCNA1) after WY-14643-induced PPARα activation mainly occurred around the PV area. This study reveals that the zonal expression of PPARα targets and proliferation-related proteins is responsible for the spatial change of hepatocyte hypertrophy and proliferation after PPARα activation. These findings provide a new insight into the understanding of PPARα activation-induced liver enlargement and regeneration.