Hepatocyte Volume Research Articles

Treatment of gestational diabetes by Acroptilon repens leaf aqueous extract green-formulated iron nanoparticles in rats

Abstract In recent years, researchers have been utilizing nanotechnology more and more to study diabetic complications, with a particular emphasis on prevention and treatment. In this investigation, we analyzed the effects of Acroptilon repens extract on iron nanoparticles (FeNPs), which demonstrated significant anti-diabetic characteristics both in living organisms and in laboratory settings. To assess the effectiveness of the FeNPs produced through the interaction of iron salt solutions stabilized by A. repens extract, we utilized a range of methodologies. The FeNPs were manufactured in a spherical shape, ranging in size from 10 to 60 nm. During the in vivo experiment, gestational diabetes was induced through streptozotocin (STZ) intraperitoneal injection. The animals were then categorized into four groups: FeNPs-60 μg/kg group, FeNPs-120 μg/kg group, normal pregnancy group, and gestational diabetes mellitus group (n = 10). FeNPs were administered intragastrically for 25 days. On the final day, the levels of ALP, AST, ALT, and blood glucose in the serum samples were assessed. Following tissue processing, 5 μm liver sections were prepared and the overall volume of the hepatic arteries, bile ducts, central vein, portal vein, sinusoids, hepatocytes, and liver, were approximated. FeNPs have the potential to reduce the elevated levels of ALP and AST enzymes. In gestational diabetes rats, the administration of FeNPs lead to a decrease in blood glucose levels. The administration of STZ significantly increased the volume of sinusoids and hepatocytes. However, after the treatment with a high dose of FeNPs, there was a notable decrease in their volume. In contrast, the volume of the bile ducts and portal vein remained unchanged in the experimental groups. Nevertheless, the volume of the hepatic arteries and central vein exhibited changes due to the presence of FeNPs. The current study showcases the hepatoprotective and anti-diabetic characteristics of FeNPs, providing a potential option as a supplement to prevent gestational diabetes mellitus while also offering hepatoprotective benefits.

Morphological characteristics of the stromal-parenchymatous component of the liver of fetal and newborn developed under the conditions of maternal hypertension disease

Background. Extragenital pathology of the mother can cause various changes in the body of the newborn, which in the future are the cause of violations in physical and mental development, and also causes an increase in somatic and infectious diseases in such children. Therefore, timely diagnosis of liver diseases in newborns and young children remains an urgent problem today. Purpose – the article is devoted to the study of the morphological features of the stromal-parenchymal component of the liver of fetuses and newborns that developed under conditions of maternal hypertension. Materials and Methods. Research was conducted on liver preparations of fetuses and newborns who died at the age of 37 weeks of gestation – the 1st day of postnatal life. All the studied material was divided into two groups. The control group (group K) included fetuses and newborns from healthy mothers (18 cases) who died due to acute disruption of umbilical-placental circulation and birth trauma. The comparison group (group C) consisted of 36 fetuses and newborns from mothers with stage II hypertension. The obtained micropreparations were stained with hematoxylin and eosin, picrofuchsin according to the van Gizon method, and according to the Mallory method. The parenchymal, stromal, and vascular components were calculated as a percentage, and then the stromal-parenchymal ratio was calculated. Results. The results we obtained during the conducted research indicated that the influence of chronic intrauterine hypoxia, which developed against the background of stage II hypertensive disease in the mother, leads to the development of destructive-dystrophic changes in the liver tissue of fetuses and newborns. Chronic intrauterine hypoxia with maternal arterial hypertension leads to parenchymal losses and has a pronounced stimulating effect on the stromal component of the liver, and this can lead to a significant decrease in the functional activity of the liver under the influence of certain triggers and increases the risk of developing liver failure and liver cirrhosis in the future. Conclusions. During our research, it was established that chronic intrauterine hypoxia, which develops against the background of stage II hypertensive disease in the mother, leads to the development of significant destructive-dystrophic changes in the liver tissue of fetuses and newborns. There is a decrease in the volume of the parenchyma due to the volume of hepatocytes and an increase in the volume of the stroma with vessels due to the volume of the portal tracts (11.2 ± 2.2% compared to the control – 3.4 ± 0.3%), the volume of the central veins (12.9 ± 1.3% compared to the control – 9.4 ± 0.4%), the volume of sinusoids (13.3 ± 1.1% compared to the control – 11.3 ± 0.8%). It was established that the morphological changes described by us in the liver of fetuses and newborns in the subsequent ontogenesis may become a prerequisite for the development of hepatobiliary system pathology in such children.

Acrylamide-Aggravated Liver Injury by Activating Endoplasmic Reticulum Stress in Female Mice with Diabetes.

Acrylamide (ACR) is a common industrial contaminant with endocrine-disrupting toxicity. Numerous studies have indicated that females and diabetics are more sensitive to environmental contaminants. However, it remains unknown whether female diabetics are susceptible to ACR-induced toxicity and its potential mechanisms. Thus, the female ACR-exposure diabetic Balb/c mice model was established to address these issues. Results showed that ACR could induce liver injury in normal mice and cause more serious inflammatory cell infiltration, hepatocyte volume increase, and fusion in diabetic mice liver. Meanwhile, ACR could lead to exacerbation of diabetic symptoms in diabetic mice by disturbing the glucose and lipid metabolism in the liver, which mainly manifests as the accumulation of liver glycogen and liver lipids, the reduction of the activity/content of glycolytic and metabolizing enzyme as well as pentose phosphatase, upregulation of the gene expression in fatty acid transporter and gluconeogenesis, and downregulation of the gene expression in fatty acid synthesis and metabolism. Moreover, ACR exposure could induce oxidative stress, inflammation, and endoplasmic reticulum stress in the liver by a decrease in hepatic antioxidant enzyme activity and antioxidant content, an increase in inflammatory factor levels, and a change in the related protein expression of endoplasmic reticulum stress (ERS) and apoptosis-related pathways in diabetic mice. Statistical analysis results revealed that ACR-induced liver injury was highly correlated with inflammation and oxidative stress, and ERS and diabetic mice had a higher risk of liver injury than normal mice. Overall results suggested that female diabetic mice easily suffer from ACR-induced toxicity, and the reason was that ACR could induce further damage to the liver by worsening the condition of inflammation, oxidative stress, and ERS in the liver.

Chemical characterization and effect of Ziziphora clinopodioides green-synthesized silver nanoparticles on cytotoxicity, antioxidant, and antidiabetic activities in streptozotocin-induced hepatotoxicity in Wistar diabetic male rats.

The present research studied the cytotoxicity, antioxidant, and antidiabetic activities of biogenically synthesized silver nanoparticles (AgNPs) using Ziziphora clinopodioides (Z. clinopodioides) as a green mediator. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), and ultraviolet-visible spectroscopy (UV-Vis) were employed to determine AgNPs. In the invivo experiment, the model rats were categorized into different groups receiving 50, 100, 200, and 400 μg/kg of AgNPs and diabetic, positive, and normal groups (n = 10) using a random division. A single dose of streptozotocin (STZ) at 60 mg/kg was administered to induce diabetes and hepatotoxicity in rats. The administration of AgNPs was performed via intragastric administration for 25 days. On the final day, the levels of glucose and biochemical enzymes, namely aspartate aminotransferase (AST), alkaline phosphatase (ALP), alanine transaminase (ALT), and gamma-glutamyltransferase (GGT), were assessed in the serum. Following tissue processing, liver sections with a thickness of 5 μm were prepared. Later, the total volume of different liver components, such as hepatocytes, sinusoids, portal vein, central vein, hepatic arteries, and bile ducts, was measured. The portal vein and bile duct volumes did not vary significantly in groups treated by AgNPs. However, the volume of the central vein and hepatic arteries exhibited noticeable variations in groups treated by AgNPs. After administration of streptozotocin, the volume of hepatocytes and sinusoids increased significantly. However, treatment with a high dose of AgNPs significantly decreased the volume of hepatocytes and sinusoids. In diabetic rats, administering AgNPs reduced the fasting blood glucose levels compared to the model group. In addition, AgNPs decreased the elevated levels of AST and ALP enzymes in a manner that depended on the dosage of AgNPs used. This research demonstrates the hepatoprotective and antidiabetic properties of AgNPs, suggesting their potential implications as hepatoprotective and antidiabetic supplements to prevent diabetes.

Quantitative morphometry of compensated liver cirrhosis can aid to predict the outcomes of surgical treatment

Objective — to define quantitative morphometric characteristics of hepatic parenchyma lesions at compensated liver cirrhosis (CLC), which will allow more accurately to assess its functional reserves and predict the outcomes of surgical treatment. Materials and methods. Intraoperative marginal liver biopsy was performed for 74 patients with CLC, who underwent surgical treatment. Morphological examinations with quantitative morphometry of intraoperative liver biopsies were done with the use of cytological analyzer with software «Integral‑2MT» produced by the association «Kvant» (Kyiv). The connective tissue area (CTA), unchanged hepatocytes area (UHA), stromal‑parenchymatous ratio (SPR), the volume of hepatocytes in the state of necrosis and/or necrobiosis (VHSNN), the volume of hepatocytes in the state of division (VHSD) were determined. The statistical analysis was performed by use of Statistica 12 software package. Results. Three types of morphological pattern of CLC were isolated. Type I (type A) portal cirrhosis with weakly pronounced signs of parenchymatous and stromal reaction (mono‑multilobular type, portal cirrhosis). The CTA was 66.73±1.71 mm2, SPR was 0.285±0.019, UHA was 234.13±11.5 mm2, VHSNN was 11.21±0.74%, VHSD was 10.23±0.57%. Type II cirrhosis (type B) — cirrhosis of mono‑multilobular type with moderately expressed signs of parenchymatous and stromal reaction. The CTA was 126.69±12.5 mm2, SPR was 0.617±0.031, UHA was 205.34±13.8 mm2, VHSNN was 17.32±0.63%, VHSD was 15.43±0.48%. Type III cirrhosis (type C) — cirrhosis with pronounced signs of parenchymatous and stromal reaction, more often of multilobular type. The CTA was 240.16±13.4 mm2, SPR was 1.344±0.089, UHA was 178.69±18.7 mm2, VHSNN was 23.97±0.75%, VHSD was 11.07±0.58%. Analysis of immediate results of surgical treatment demonstrated, that no one patient with A‑type CLC died, while 9.5% of patients with B‑type and 25.0% of patients with C‑type CLC died in the early postoperative period. Conclusions. Quantitative morphometry demonstrated that pathological changes in the liver at CLC are heterogeneous and can be categorized in to 3 types (A, B, C). Three types of morphological pattern at CLC, which were isolated, are characterized by significant differences in CTA, UHA, SPR, VHSNN, VHSD parameters. At transition of A‑type cirrhosis into C‑type, volume of hepatic parenchyma becomes to be decreased, while volume of connective tissue becomes to be increased. This is accompanied by decrease in UHA, increase in CTA, SPR and VHSNN. Surgical intervention in patients with type C morphological pattern of cirrhosis is associated with a high risk (25%) of poor outcome, which indicates the need to limitations for indications to elective surgical operations in this category of patients. Thus, quantitative morphometry of liver biopsies in CLC patients can aid to predict the outcomes of surgical treatment.

Effect of aqueous-alcoholic extract of Ducrosia Anethifolia Boiss on the fetal liver of diabetic rats

Objective: Moshgak (Ducrosia anethifolia) is a wild plant with medicinal value. The present study aimed to evaluate the effect of Moshgak on the liver tissue of the diabetic rat fetus. Materials and Methods: In this animal study, the aqueous-alcoholic extract of Moshgak was prepared in the standard method. Forty rats were divided randomly into five groups, including control, sham, and three diabetic groups. The rats were diabetic with intraperitoneal injection of a single dose of streptozotocin (80 mg) and 2 diabetic groups were treated with Moshgak extract (280 and 560 mg/kg/bw) for 19 days. The rats were anesthetized and their blood was taken to measure the blood glucose, insulin, and malondialdehyde. Then, their fetuses were removed. The fetal liver sections were obtained by using the stereological methods. The micrometry of the liver tissue was performed and data were analyzed. Results: The finding showed a statistically significant increase (P < 0.01) in the total volume of liver, connective tissue, sinusoid, and hepatocytes in diabetic rats compared to control rats, while these parameters decreased significantly in treated groups with Moshgak. Hepatic cell count hepatic decreased in the treated groups. Furthermore, the changes in blood glucose, malondialdehyde, and insulin in diabetic rats were improved significantly by Moshgak treatment. The dilation of sinusoids, hepatocyte vacuolation, and mild lymphocytosis was observed in all diabetic groups except the treatment group with Moshgak 560 mg/kg/bw. Conclusions: According to obtained results, Moshgak extract was able to compensate partially the changes induced by diabetes in the fetal liver tissue. Therefore, due to the side effects of diabetes during pregnancy, further research on anti-diabetic properties of Moshgak is suggested.

Comparative study of carbohydrate levels on growth, oxidative stress and glucolipid metabolism of hybrid fish between Megalobrama amblycephala (♀) × Culter alburnus (♂) and Culter alburnus

The present study used hybrid fish (Megalobrama amblycephala ♀ × Culter alburnus ♂, abbreviated as BT) and topmouth culter (Culter alburnus, abbreviated as TC) as research subjects to assess and compare the effects of carbohydrate on the growth, oxidative stress and glucolipid metabolism of the two fish. The TC (initial weight, 13.10 ± 0.10 g) and BT (initial weight, 12.85 ± 0.13 g) were fed isonitrogenous (43% crude protein) and isolipidic (6.5% crude lipid) diets with carbohydrate levels of 10% (LC diet) and 30% (HC diet) for 11 weeks, respectively. The results showed that the HC diet resulted in a significant decrease in specific growth rate (SGR) of both fish. However, the SGR in BT was significantly higher than in TC, regardless of diet. Furthermore, HC diet significantly increased serum alanine transaminase (GPT) and hepatic malondialdehyde (MDA) content, decreased hepatic catalase (CAT) activity and resulted in increased hepatocyte volume, nucleus deficiency and nucleus deviation from the center of cytoplasm in both fish. However, BT had significantly higher glutathione peroxidase (GPx) activity and lower serum GPT and aspartate transaminase (GOT) content compared to TC. The HC diet induced significant increases in hepatosomatic index (HSI), liver lipid droplet area, liver glycogen, serum glucose (GLU) and triglyceride (TG) content in both fish. However, HSI and lipid droplet area were significantly higher in BT than in TC, while serum GLU and TG content were significantly lower in BT than in TC. Both fish can adapt to the HC diet by up-regulating the expression of glycolysis-related gene (glucokinase (gk)) and lipogenesis-related genes (acetyl-CoA carboxylase a (acca), fatty acid synthase (fas)). However, the expression levels of glucose transporter 2 (glut2), glycolysis-related gene (phosphofructokinase liver b (pfklb)) and lipogenesis-related genes (sterol regulatory element-binding protein 1 (srebp1), ATP citrate lyase a (aclya)) were increased only in BT fed HC diet. In summary, the BT liver has stronger glucose transport, glycolysis, and lipid tolerance compared to TC, thus exhibiting better glucose homeostasis and growth performance. It indicates that hybridization may help to produce new strain of fish with high carbohydrate utilization capacity.

Prolonged Normothermic Machine Perfusion: Buying More Time for Liver Graft Assessment and Repair.

Prolonged Normothermic Machine Perfusion: Buying More Time for Liver Graft Assessment and Repair.

Transcriptome analysis reveals the host immune response upon LMBV infection in largemouth bass (Micropterus salmoides)

Largemouth bass (Micropterus salmoides) is one of the important economical freshwater aquaculture species in China. However, the outbreak of viral diseases always caused great economic losses in the largemouth bass aquaculture industry. Largemouth bass virus (LMBV), a double-stranded DNA (dsDNA) virus belonging to genus Ranavirus, family Iridoviridae causes high mortality in cultivated largemouth bass. However, host responses, especially the molecular events involved in LMBV infection still remained largely uncertain. Here, we established an in vivo model of LMBV infection, and systematically investigated the mRNA expression profiles of host genes in liver and spleen from infected largemouth bass using RNA sequencing (RNA-seq). Histopathological analysis indicated that necrotic cells and the formed necrotic focus were present in spleen, while numerous basophilic cells, hepatocytes volume shrinkage, nucleus pyknosis, and the disappeared boundary of hepatocytes were observed in the liver of infected largemouth bass. Transcriptomic analysis showed that transcription levels of 5128 genes (2804 up-regulated genes and 2324 down-regulated) in liver and 7008 genes (2603 up-regulated and 4405 down-regulated) in spleen were altered significantly. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis indicated that numerous co-regulated differentially expressed genes (DEGs) in liver and spleen were enriched in the pathways related to cell death and immune signaling, such as apoptosis, necroptosis, cytokine-cytokine receptor interaction and JAK-STAT signaling. Moreover, the DEGs specially regulated by LMBV infection in liver were significantly enriched in the KEGG pathways related to metabolism and cell death, while those in spleen were enriched in the immune related pathways. In addition, the expression changes of several randomly selected genes, such as SOCS1, IL-6, CXCL2, CASP8, CYC and TNF from qPCR were consistent with the transcriptomic data. Taken together, our findings will provide new insights into the fundamental patterns of molecular responses induced by LMBV in vivo, but also contribute greatly to understanding the host defense mechanisms against iridoviral pathogens.

Effects of a high-carbohydrate high-fat diet on liver morphology in young and old rats

The liver is a complex metabolic organ that, through the regulation of energy metabolism, maintains the homeostasis of the whole organism.Aim: To establish the morphological changes in hepatocytes and the expression of endothelial vascular factor in a high-carbohydrate high-fat diet (HCHFD) depending on age.Material and Methods. A histological study of the liver was carried out in four groups of male Wistar rats: 1st group – 5-month-old animals on a standard diet; 2nd group – 5-month-old animals on HCHFD for 90 days (from 2 months of age); 3rd group – 18-month-old animals on a standard diet; 4th group – 18-month-old animals on HCHFD for 90 days (from 15 months of age). Using morphometric methods, the specific volumes (%) of unchanged and binuclear hepatocytes, sinusoidal capillaries, inflammatory infiltrates, and foci of fibrosis in the liver were determined. The expression of vascular endothelial growth factor (VEGF) in endothelial cells and hepatocytes was evaluated by immunohistochemical method. The concentration of cholesterol and triglycerides was determined by the enzymatic colorimetric method in the blood serum and in the liver.Results. A morphological study revealed a significant expansion of sinusoidal capillaries in groups 2 and 4. A statistically significant increase in the specific volume of hepatocytes with vacuolar inclusions, binuclear hepatocytes, fibrous foci and small inflammatory infiltrates was detected in the 4th group. VEGF expression increased in hepatocytes of groups 2 and 4. Biochemical study found an increase in the concentration of triglycerides in the rats’ liver of the 4th group. Thus, HCHFD, despite the pronounced signs of regeneration, increased age-related changes in the liver of old rats.

Morphological changes in the liver of rats after administration of chlorpromazine, depending on the dose and duration of administration

Chlorpromazine (CPZ) remains a widely used drug in psychiatric practice today. The drug has a hepatotoxic effect, but the possible mechanisms of this side effect have not yet been fully elucidated. The aim of the study was to determine morphological changes in rat liver tissue under chronic toxic effects of chlorpromazine, depending on the dose and duration of its administration. The study was conducted on 60 sexually mature male rats. CPP was administered intragastrically at different doses (3.5, 7.0, 14.0 and 21.0 mg/kg) for 30 and 60 days. The material was fixed in a 10 % solution of neutral formalin (pH 7.2-7.4) for 24-48 hours, then passed through alcohols of increasing concentration and embedded in paraffin. Serial sections (6-7 µm thick) were prepared from the paraffin blocks and stained with hematoxylin-eosin and picrofuchsin by Van Gieson to determine the degree of fibrotic changes in liver tissue, as well as with Giemsa III to detect fatty degeneration of hepatocytes. The microscopic structure of the hepatic parenchyma was studied using an OLIMPUS BX41 light microscope at 100, 200 and 400x magnification. Morphometric parameters of structural changes were determined using an ocular grid and Image Tulsa 3.6 software. The data were statistically processed by descriptive statistics using the Microsoft Office Excel 2010 spreadsheet processor. When CPZ was administered in different doses and duration, pathological changes of varying severity developed in the liver tissue of rats. In the liver tissue, signs of intracellular and intra-tubular cholestasis are found mainly in the central lobes, accompanied by focal desquamation and proliferation of the biliary epithelium, formation of small-focal, less frequently zonal necrosis of hepatocytes, inflammatory infiltration of portal tracts with its spread to the interlobular stroma and parenchyma. Mitotically active binucleated hepatocytes are the key to the reparative process. Periductal fibrosis develops in the portal sections, marginal proliferation of the bile ducts, hepatocytes with signs of granular and/or fatty dystrophy are noted. In the central veins and vessels of the portal areas, moderate initial sclerotic changes were found, signs of their capillarisation in sinusoids, and the endothelium of the vessels had focal destructive changes. In all portal zones, proliferation of bile ducts and formation of bile pseudo-ducts were observed. Thus, the analysis of the morphometric study data showed that within 60 days of CPZ administration there is a significant increase in the relative volume of connective tissue and stromal-parenchymal index due to a significant decrease in the volume of hepatocytes.

Automated image analysis of keratin 7 staining can predict disease outcome in primary sclerosing cholangitis.

Primary sclerosing cholangitis (PSC) is a chronic cholestatic liver disease that obstructs the bile ducts and causes liver cirrhosis and cholangiocarcinoma. Efficient surrogate markers are required to measure disease progression. The cytokeratin 7 (K7) load in a liver specimen is an independent prognostic indicator that can be measured from digitalized slides using artificial intelligence (AI)-based models. A K7-AI model 2.0 was built to measure the hepatocellular K7 load area of the parenchyma, portal tracts, and biliary epithelium. K7-stained PSC liver biopsy specimens (n=295) were analyzed. A compound endpoint (liver transplantation, liver-related death, and cholangiocarcinoma) was applied in Kaplan-Meier survival analysis to measure AUC values and positive likelihood ratios for each histological variable detected by the model. The K7-AI model 2.0 was a better prognostic tool than plasma alkaline phosphatase, the fibrosis stage evaluated by Nakanuma classification, or K7 score evaluated by a pathologist based on the AUC values of measured variables. A combination of parameters, such as portal tract volume and area of K7-positive hepatocytes analyzed by the model, produced an AUC of 0.81 for predicting the compound endpoint. Portal tract volume measured by the model correlated with the histological fibrosis stage. The K7 staining of histological liver specimens in PSC provides significant information on disease outcomes through objective and reproducible data, including variables that cannot be measured by a human pathologist. The K7-AI model 2.0 could serve as a prognostic tool for clinical endpoints and as a surrogate marker in drug trials.

Liver comparative transcriptome analysis reveals the mechanism of the Hainan medaka, Oryzias curvinotus, to adapt to salinity

AbstractTo explore the adaptation mechanism of Hainan medaka, Oryzias curvinotus, to salinity, this study performed a histological and RNA‐Seq comparative analysis of female medaka liver with different salinities (0, 15 and 30 practical salinity unit [psu]). In the 30 psu group, the volume of hepatocytes was increased and cytoplasmic fat accumulated. A total of 83,758 unigenes were assembled by RNA‐Seq, of which 31,147 were annotated. In total, 922 differentially expressed genes (DEGs) were obtained, and the detection results of 24 DEGs by quantitative polymerase chain reaction (qPCR) were consistent with the RNA‐Seq data. Gene ontology (GO) enrichment showed that DEGs were mainly involved in biological processes, such as substance synthesis and transport, energy metabolism, signal transduction, and had catalytic activity and binding functions. Kyoto Encyclopedia of Genes and Genomes enrichment analysis showed that DEGs were significantly enriched in steroid biosynthesis, glycerophospholipid metabolism, peroxisome proliferator‐activated receptor signaling pathway, and insulin signaling pathway that regulate lipid metabolism. The results of this study highlight the response of the liver transcriptome to long‐term environmental salinity changes and provide a basis for the application of the medaka genetic resources.

Responses of Wild Finescale Dace (Phoxinus neogaeus) to Experimental Spills of Cold Lake Blend Diluted Bitumen at the International Institute for Sustainable Development-Experimental Lakes Area, Northwestern Ontario.

Pipelines carrying diluted bitumen (dilbit) traverse North America and may result in dilbit release into sensitive freshwater ecosystems. To better understand the potential effects of a freshwater oil release, the Boreal-lake Oil Release Experiment by Additions to Limnocorrals project at the International Institute for Sustainable Development-Experimental Lakes Area (Ontario, Canada) modeled seven dilbit spills contained within a 10-m diameter of littoral limnocorrals in a boreal lake. Wild finescale dace (Phoxinus neogaeus) were released in the limnocorrals 21 days after oil addition and remained there for 70 days. Dilbit volumes covered a large range representing a regression of real spill sizes and total polycyclic aromatic compounds (TPAC) between 167 ng L-1 day-1 and 1989 ng L-1 day-1 . We report the effects of chronic exposure on reproductive potential as well as physiological responses in the gallbladder and liver. In exposures >1000 ng L-1 day-1 , there was a significant decrease in fish retrieval, culminating in zero recapture from the three highest treatments. Among the fish from the limnocorrals with lower levels of TPAC (<500 ng L-1 day-1 ), effects were inconsistent. Gallbladder bile fluorescence for a naphthalene metabolite was significantly different in fish from the oil-exposed limnocorrals when compared to the lake and reference corral, indicating that fish in these lower exposures were interacting with dilbit-derived polycyclic aromatic compounds. There were no significant differences in condition factor, somatic indices, or hepatocyte volume indices. There were also no significant changes in the development of testes or ovaries of exposed dace. The results from the present study may serve to orient policymakers and emergency responders to the range of TPAC exposures that may not significantly affect wild fish. Environ Toxicol Chem 2022;41:2745-2757. © 2022 SETAC.

Effect of Aerobic Exercise on Lipid Metabolism in Rats With NAFLD.

This work aimed to study the intervention effect of exercise on lipid metabolism in NAFLD rats, provide a more scientific experimental basis for exploring and improving the theoretical system of exercise intervention in NAFLD, and further provide a theoretical research basis for clinical treatment of NAFLD. Forty healthy male Sprague Dawley rats were randomly divided into a blank control group (BC,14) and a model group (MO, 26). After 6°weeks of modeling, the MO group was randomly divided into the model control group (MC, 12) and the aerobic exercise group (AE, 12). Platform running intervention in group E was conducted at a slope of 0°, a speed of 15 m/min, 1 h/time, once a day, six times a week, and a day of rest, for 8°weeks in total. After the intervention, the liver tissues of rats were taken for pathological sections, and serum was taken and analyzed for TC, TG, LDL-C, HDL-C, and FFA levels. Under the light microscope, the liver tissue structure of rats in the BC group was complete and clear, the structure of liver lobules was clear and normal, the volume of hepatocytes was uniform, the nucleus was in the middle, and the cytoplasm was red-stained, and no steatosis of hepatocytes was found. The liver of rats in the MC group showed diffuse fatty lesions, disordered structure of hepatic lobules, disordered arrangement of hepatic cords, different sizes of hepatocytes, loose cytoplasm, and diffuse lipid droplets of different sizes in the cytoplasm. The accumulation of liver lipid droplets in the AE group was improved compared with the MC group, the number of fat vacuoles in hepatocytes was significantly reduced, and the degree of liver lipid deposition was reduced. Compared with the BC group, the content of TC, TG, LDL-C, and FFA in the serum of the MC group increased significantly (p < 0.01), and the content of HDL-C decreased significantly (p < 0.01). Compared with the MC group, the content of TC, TG, LDL-C, and FFA in the serum of the AE group decreased significantly (p < 0.01/p < 0.05), and the content of HDL-C increased significantly (p < 0.01). Therefore, moderate-intensity aerobic exercise has an intervention effect on lipid metabolism in NAFLD rats, which can be used as one of the means to treat NAFLD.

Investigation of Diazepam Sequential Metabolism with Hepatocytes via a Microfluidic Device

Objectives and hypothesisThe purpose of this study is to investigate the sequential metabolism of diazepam (DZP) in a dynamic system. In rat liver, DZP can be metabolized into primary metabolite temazepam (TZP), nordiazepam (NDP), p‐hydroxydiazepam (PHD), and secondary metabolite oxazepam (OXP) and temazepam glucuronide (T‐G). It is reported that a lag time of DZP metabolite formation can be observed in rat liver single‐pass perfusion and PK study. We are using a microfluidic device to represent liver sinusoid and to investigate the contribution of metabolism and drug transport in the disposition of diazepam and its metabolite along the sinusoid. Ordinary differential equation (well‐stirred model) and partial differential equation (PDE) models are being developed to characterize the disposition of DZP and its metabolites. We hypothesize that characterization of sequential metabolism in a dynamic system compared to a static system (microsomal incubation or static hepatocyte incubation) can provide a more accurate prediction of in vivo sequential metabolism.MethodsThe perfusion study was conducted with rat hepatocytes. Rat hepatocytes were seeded in collagen I coated microscopy slides to form a monolayer, followed by three‐day enzyme induction. An ibidi® μ‐sticky slide was mounted on the microscopy slide to create a microfluidic device. Hepatocyte culture medium containing 5 μM DZP was perfused through the device and subsequently perfused by DZP‐free culture medium. The concentration of diazepam and its metabolites in the perfusate was measured by LC/MS/MS. Mathematical modeling was applied to predict the concentration‐time (C‐t) profile of the perfusion study via Mathematica version 12.3 (software). The model parameters include metabolic intrinsic clearance (CLint,met) and passive diffusion clearance (CLdiff), partitioning into membranes, hepatocyte volume, and device volume. Other relevant model parameters were collected by literature search.ResultsAt 10 μL/min flow rate, three primary metabolites all show similar lag time (pre device dead volume plus device volume) to that of DZP. OXP shows a longer lag time compared to T‐G. The lag time of T‐G is similar to that of primary metabolites. However, a significant decline of TZP and NDP formation at a steady‐state is observed, but not PHD. One possible rationale is constant enzyme loss for CYP3A (responsible for TZP, NDP formation), but not CYP2D (responsible for PHD formation). For the DZP‐free perfusion phase, a similar lag time was observed for DZP and its metabolites. A well‐stirred model and a PDE model to characterize the dimension and spatial information are being optimized.ConclusionsGenerally, this microfluidic device can characterize the sequential metabolism of diazepam. The C‐t profiles of DZP and its metabolites are similar to that of the liver single‐pass perfusion study. Due to the existence of transporters, the lag time of T‐G is similar to that of primary metabolites. As a future study, the distribution of DZP and its metabolites along the device will be measured by MALDI imaging. The information of spatial distribution is expected to further help optimize and validate the mathematical models.

Lymphocyte-Specific Protein-1 Suppresses Xenobiotic-Induced Constitutive Androstane Receptor and Subsequent Yes-Associated Protein–Activated Hepatocyte Proliferation

Activation of constitutive androstane receptor (CAR) transcription factor by xenobiotics promotes hepatocellular proliferation, promotes hypertrophy without liver injury, and induces drug metabolism genes. Previous work demonstrated that lymphocyte-specific protein-1 (LSP1), an F-actin binding protein and gene involved in human hepatocellular carcinoma, suppresses hepatocellular proliferation after partial hepatectomy. The current study investigated the role of LSP1 in liver enlargement induced by chemical mitogens, a regenerative process independent of tissue loss. 1,4-Bis [2-(3,5-dichloropyridyloxy)] benzene (TCPOBOP), a direct CAR ligand and strong chemical mitogen, was administered to global Lsp1 knockout and hepatocyte-specific Lsp1 transgenic (TG) mice and measured cell proliferation, hypertrophy, and expression of CAR-dependent drug metabolism genes. TG livers displayed a significant decrease in Ki-67 labeling and liver/body weight ratios compared with wild type on day 2. Surprisingly, this was reversed by day 5, due to hepatocyte hypertrophy. There was no difference in CAR-regulated drug metabolism genes between wild type and TG. TG livers displayed increased Yes-associated protein (YAP) phosphorylation, decreased nuclear YAP, and direct interaction between LSP1 and YAP, suggesting LSP1 suppresses TCPOBOP-driven hepatocellular proliferation, but not hepatocyte volume, through YAP. Conversely, loss of LSP1 led to increased hepatocellular proliferation on days 2, 5, and 7. LSP1 selectively suppresses CAR-induced hepatocellular proliferation, but not drug metabolism, through the interaction of LSP1 with YAP, supporting the role of LSP1 as a selective growth suppressor.

Microcystin-LR at sublethal concentrations induce rapid morphology of liver and muscle tissues in the fish species Astyanax altiparanae (Lambari)

The process of eutrophication and consequent proliferation of cyanobacteria in rivers and lakes leads to increasing numbers of harmful algal blooms and higher concentration of toxic metabolites in freshwater bodies. Microcystin is a toxic metabolite produced by cyanobacteria that is frequently detected and can pose health risks to important freshwater species including fish. The aim of the present study was to evaluate the effects of microcystin-LR on the morphology of Astyanax altiparanae's liver and muscle. One hundred (n = 100) Astyanax altiparanae were divided into 5 groups (n = 20) with 24 h and 96 h of microcystin exposition at two doses of 0.5 and 1.0 μg/L. Differences were observed in the microcystin treatment with respect to histopathological analyses including cytoplastic degradation, displacement, and increase in nuclei volume and area of hepatocytes. Hyperemia and dilation of blood capillaries were seen in the liver. There were also observable changes in the size of muscle fibers and muscle inflammation. Our results demonstrate that microcystins can impact the integrity of both tissues even at sublethal concentrations. Low doses of microcystins are therefore sufficient to intoxicate fish livers and muscle tissues.

Тканевый и ультраструктурный анализ печени крыс препубертатного возраста при субтоксическом воздействии кадмия и свинца

Using light and electron microscopy, the effect of intoxication with cadmium sulfate (Cd) and lead acetate (Pb) for 3 weeks on the architectonics of the liver parenchyma and ultrastructural features of hepatocytes of 40 prepubertal male Wistar rats was studied, taking into account the model of the classical hepatic lobule. Animals were divided into 4 groups: 1st – control; 2nd – cadmium sulfate solution at a dose of 0.5 mg/kg; 3rd – lead acetate solution 10 mg/kg; 4th – a combination of these solutions. According to stereological analysis of semi-thin sections, statistically significant indicators of periportal zones reflect an increase in the volume of hepatocytes when exposed to Cd (2nd and 4th groups). In all groups, with the exception of the 3rd one (Pb), an increase in the parenchymal compartment in the porto-central gradient was revealed, indicating the formation of hepatocellular ballooning. In all experimental groups, for the periportal zone of the hepatic lobule, a characteristic feature of the ultrastructure of hepatocytes was a statistically significant decrease in the volumetric density of the endoplasmic reticulum; for the centrilobular zone – of mitochondria; for the perivenular zone – mitochondria and nuclei of hepatocytes. The formation of autophagosomes with signs of predominantly mitochondrial utilization attracted attention in groups exposed to Pb. Pathological changes are compensated by increased transsinusoidal metabolism and complexes of biosynthesis organelles.

Liver function quantification of patients with portal vein embolization using dynamic contrast-enhanced MRI for assessment of hepatocyte uptake and elimination

PurposeWe evaluated pharmacokinetic models which quantify liver function including biliary elimination based on a dynamic Gd-EOB-DTPA-enhanced magnetic resonance imaging (MRI) technique with sparse data collection feasible in clinical routine. MethodsTwelve patients with embolized liver segments following interventional treatment of primary liver cancer or hepatic metastasis underwent MRI. During Gd-EOB-DTPA bolus administration, a 3D dynamic gradient-echo (GRE) MRI examination was performed over approx. 28 min. Interrupted data sampling was started approx. 5 min after contrast agent administration. Different implementations of dual-inlet models were tested, namely the Euler method (DE) and convolution with residue functions (C). A simple uptake model (U) and an uptake- elimination model (UE) extended by incorporating the biliary contrast agent elimination rate (Ke) were evaluated. ResultsThe uptake-elimination model, calculated via the simple Euler method (UE- DE) and by convolution (UE-C), yielded similar overall estimates in terms of fitting quality and agreement with published values. The Euler method was approx. 50 times faster and yielded a mean elimination rate of Ke=1.8±1.2mL/(min·100 mL) in nonembolized liver tissue, which was significantly higher (p=8.8·10-4) than in embolized tissue Ke=0.4±0.4 mL/(min·100 mL). Fractional hepatocyte volume vh was not significantly higher in nonembolized tissue (52.4 ± 13.4 mL/100 mL) compared to embolized tissue (44.4 ± 26.1 mL/100 mL). ConclusionsInterrupted late enhancement MRI data sampling in conjunction with the uptake-elimination model, deconvolved by integration of the differential rate equation and combined with the simple uptake model implemented with the Euler method (U-DE), turned out to be a stable and practical method for reliable noninvasive assessment of liver function.