Rat Liver Sections Research Articles

Exposure to a choline-deficient diet during pregnancy and lactation alters the liver transcriptome profile in offspring of dams with fatty liver.

The developmental origin of health and disease hypothesis shows that early adverse exposures can have lifelong health effects. Thus, the aim of this study was to analyze the impact of choline intake during pregnancy and/or lactation on gene expression profiles in the liver of 24-day-old male rat offspring from dams with non-alcoholic fatty liver disease (NAFLD). Phenotypic characteristic, histological examination and global transcriptome pattern of liver tissue specimens obtained from offspring of dams suffering from fatty liver, provided with proper choline intake during pregnancy and lactation (NN), fed a choline-deficient diet during both periods (DD), deprived of choline only during pregnancy (DN), or only during lactation (ND), was performed. The global gene expression profile was analyzed by using microarray approach (Affymetrix® Rat Gene 2.1 ST Array Strip). The relative expression of selected genes was validated by real-time polymerase chain reaction (qPCR). The histological examination of rat liver sections indicated alternations typical for fatty liver in all analyzed groups with increased progression among groups deprived of choline. Choline deficiency in the maternal diet was associated with changes in body mass and composition but not with biochemical marker levels, except for the high density lipoprotein fraction of cholesterol (HDL). Enhanced expression of genes involved in oxidative stress, cell proliferation, activation of catabolic processes related to hepatocyte dysfunction and cell membrane composition were simultaneously observed in all choline-deficient groups. An adequate amount of choline in the diet of a mother with fatty liver during pregnancy and/or lactation can regulate gene expression in the offspring's liver and contribute to a milder stage of the disease in the progeny. Moreover, proper choline supply during the postpartum period is as crucial as during the prenatal period.

Histopathological Studies on the Effect of Taurine Against Etoposide-Induced Acute Liver and Kidney Toxicity in Female Albino Rats

Etoposide (ETP) is a topoisomerase Ⅱ (TOP Ⅱ) inhibitor and one of the leading chemotherapeutic drugs for treating a wide variety of tumors. However, ETP induced hepatotoxicity and nephrotoxicity limits its clinical use. This study aims to investigate the protective potential of taurine (Tau) to mitigated histopathological changes in the liver and kidneys of female albino rats treated with ETP. A total of 18 female rats were divided into three groups; control group, ETP-exposed group received intraperitoneal injection of ETP on the first 3 days of the study for a total cumulative dose of 44 mg/kg to induce hepatotoxicity and nephrotoxicity, ETP + Tau group received ETP as stated previously with 400 mg/kg/day of Tau via oral gavage for 15 days. Sections from the liver and kidney were evaluated under light microscope and ETP-exposed group revealed vascular congestion, chronic inflammatory cell infiltration predominantly lymphocytes, edema, vacuolar degeneration, atypical cells, pyknosis, and necrosis while liver and kidney sections of rats treated with combination of ETP + Tau group exhibited marked alleviation of the histopathological damage. The study concludes that treatment with ETP induced marked structural damages in the liver and kidney and such morphological damages are effectively diminished by administering Tau.

Single and Multiplex Immunohistochemistry to Detect Platelets and Neutrophils in Rat and Porcine Tissues.

Platelet–neutrophil complexes (PNCs) occur during the inflammatory response to trauma and infections, and their interactions enable cell activation that can lead to tissue destruction. The ability to identify the accumulation and tissue localisation of PNCs is necessary to further understand their role in the organs associated with blast-induced shock wave trauma. Relevant experimental lung injury models often utilise pigs and rats, species for which immunohistochemistry protocols to detect platelets and neutrophils have yet to be established. Therefore, monoplex and multiplex immunohistochemistry protocols were established to evaluate the application of 22 commercially available antibodies to detect platelet (nine rat and five pig) and/or neutrophil (four rat and six pig) antigens identified as having potential selectivity for porcine or rat tissue, using lung and liver sections taken from models of polytrauma, including blast lung injury. Of the antibodies evaluated, one antibody was able to detect rat neutrophil elastase (on frozen and formalin-fixed paraffin embedded (FFPE) sections), and one antibody was successful in detecting rat CD61 (frozen sections only); whilst one antibody was able to detect porcine MPO (frozen and FFPE sections) and antibodies, targeting CD42b or CD49b antigens, were able to detect porcine platelets (frozen and FFPE and frozen, respectively). Staining procedures for platelet and neutrophil antigens were also successful in detecting the presence of PNCs in both rat and porcine tissue. We have, therefore, established protocols to allow for the detection of PNCs in lung and liver sections from porcine and rat models of trauma, which we anticipate should be of value to others interested in investigating these cell types in these species.

Histopathological and immunological study of rats liver hydatid cysts isolated from human, sheep, goat and cows.

In endemic places, liver hydatidosis is a life-threatening health issue. Many consequences such as hepatomegaly, infiltration with inflammatory cells and histopathological changes might arise as a result of liver hydatidosis. This study aimed to look into pathogenic changes in the livers of the rats that experimentally infected with hydatid cysts. These hydatid cysts were isolated from naturally infected humans, sheep, goats and cows. Liver hydatid cysts were collected from the main abattoir of Al-Muthanna province while human hydatid cysts were collected from Al-Hussein Teaching Hospital in Al-Muthanna province. The hydatid cysts were grossly and histology examined for inspection of hydatid cysts. The in vivo experiments were done by injection of hydatid protoscoleces or sand (fluid) in rats intraperitoneally. The results showed that the gross signs were same in all infected livers which range from paleness, hepatomegaly, hemorrhage and calcification. The hydatid cysts isolated from sheep and goat livers were highly fertile compared to others isolated from humans and cows. Injection of hydatid protoscoleces and hydatid fluid isolated from sheep and goat livers in rats induced the highest immune response compared to that isolated from humans and cows. The liver sections of rats that were injected with human, goat and sheep hydatid protoscoleces and fluids showed hyperplasia in the bile duct, aggregation nonnuclear cells with congested blood vessels. While liver sections of rats were received goat hydatid fluid, cow hydatid protoscoleces and hydatid fluid showed normal liver tissue. These findings suggested that the immunogenicity of hydatid materials is different according to the host and the component of hydatid cysts.

Evaluation of the toxicity profile of Bridelia ferruginea methanol stem bark extract

The present research was carried out to investigate the toxicity profile of Bridelia ferruginea methanol stem bark extract. Acute toxicity study was carried out in mice using the modified Lorke’s method to determine median lethal dose (LD50). For the subacute toxicity study, animals used were albino rats while extract was administered at dose levels of 100, 1000 and 2500 mg/kg daily for a period of 28 days. Blood samples were taken at the end of the study period for estimation of biochemical and haematological parameters. Histopathological examination of sections of the liver and kidney was also carried out. No mortality was recorded in acute toxicity study and also no any change in general behaviour of rats was observed up to dose of 5000 mg/kg. Similarly, daily administration of the stem bark extract for 28 days did not produce significant changes in body weight, haematological and biochemical parameters of treated rats across all doses except at dose of 2500 mg/kg where significant change was observed in the parameters studied when compared with values obtained from the control group. However, slight histopathological changes were observed in the liver and kidneys sections of rats treated with the methanol stem bark at dose of 1000 mg/kg and 2500 mg/kg. In conclusion, based on results of acute and subacute toxicity study studies, the methanol stem bark extract of Bridelia ferruginea may be considered safe for medicinal use especially at lower doses.

Multiscale generative model using regularized skip-connections and perceptual loss for anomaly detection in toxicologic histopathology

BackgroundAutomated anomaly detection is an important tool that has been developed for many real-world applications, including security systems, industrial inspection, and medical diagnostics. Despite extensive use of machine learning for anomaly detection in these varied contexts, it is challenging to generalize and apply these methods to complex tasks such as toxicologic histopathology (TOXPATH) assessment (i.e.,finding abnormalities in organ tissues). In this work, we introduce an anomaly detection method using deep learning that greatly improves model generalizability to TOXPATH data. MethodsWe evaluated a one-class classification approach that leverages novel regularization and perceptual techniques within generative adversarial network (GAN) and autoencoder architectures to accurately detect anomalous histopathological findings of varying degrees of complexity. We also utilized multiscale contextual data and conducted a thorough ablation study to demonstrate the efficacy of our method. We trained our models on data from normal whole slide images (WSIs) of rat liver sections and validated on WSIs from three anomalous classes. Anomaly scores are collated into heatmaps to localize anomalies within WSIs and provide human-interpretable results. ResultsOur method achieves 0.953 area under the receiver operating characteristic on a real-worldTOXPATH dataset. The model also shows good performance at detecting a wide variety of anomalies demonstrating our method’s ability to generalize to TOXPATH data. ConclusionAnomalies in both TOXPATH histological and non-histological datasets were accurately identified with our method, which was only trained with normal data.

BIOCHEMICAL AND HISTOPATHOLOGICAL EFFECTS OF HYDROETHANOLIC EXTRACT OF IRVINGIA WOMBOLU KERNELS ON RAT LIVER AND KIDNEY

Objective: Many plants are consumed as food by humans for growth and survival, but a large number of these plants have not been tested for toxicity potential. Repeated consumption of such plants could lead to accumulation of toxic chemicals in the body and cause health-related problems. Irvingia wombolu kernel is widely consumed by many ethnic groups in Nigeria and some other African countries. The toxicity potentials of Irvingia wombolu kernel extract (IWKE) on the kidney and liver of rats was evaluated in this study. Methods: Three groups of Wistar rats were fed orally with IWKE (50, 100, and 200 mg/kg b.w) daily for 28 days. The fourth group which is the control was treated with distilled water (10 ml/kg b.w) for the same period. The rats were sacrificed on the 29th day, and blood samples, kidney and liver were harvested for analyses. Serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein, albumin, bilirubin, creatinine, and urea were determined. Kidney and liver sections were examined for histopathological changes. Data were subjected to Student’s t-test for statistical analysis. Results: Serum levels of creatinine and urea were not significantly altered in the IWKE-treated rats compared with the control. At 200 mg/kg b.w, the extract significantly increased (p<0.05) serum levels of ALT, AST, total bilirubin, total protein, and albumin compared with the control. Significant distortions were observed in the liver sections of rats treated with 200 mg/kg b.w IWKE compared with control, but the structure of the kidney section of IWKE-treated rats was not significantly different from the control. Conclusion: The results showed that repeated ingestion of Irvingia wombolu kernel at a dose of 200 mg/kg b.w for 28 days induced liver damage, but does not significantly affect renal function. Peer Review History: Received: 9 September 2021; Revised: 12 October; Accepted: 27 October, Available online: 15 November 2021 Academic Editor: Dr. Ali Abdullah Al-yahawi, Al-Razi university, Department of Pharmacy, Yemen, alyahawipharm@yahoo.com UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.5/10 Reviewers: Dr. Sangeetha Arullappan, Universiti Tunku Abdul Rahman, Malaysia, sangeetha@utar.edu.my Ahmad Najib, Universitas Muslim Indonesia, Makassar, Indonesia, ahmad.najib@umi.ac.id Taha A.I. El Bassossy, Medicinal and Aromatic Plants Department, Desert Research Center, Cairo, Egypt, tahachemist2008@gmail.com Dr. Nazim Hussain, North East Frontier Technical University, Arunachal pradesh, India, nhussain116@gmail.com Similar Articles: MYOCARDIAL POTENCY OF AQUEOUS EXTRACT OF HARUNGANA MADAGASCARIENSIS STEM BARK AGAINST ISOPROTERENOL-INDUCED MYOCARDIAL DAMAGE IN RATS PROTECTIVE EFFECT OF METHANOL EXTRACT OF RUSSELIA EQUISETIFORMIS AGAINST PARACETAMOL-INDUCED HEPATOTOXICITY IN WISTAR RATS

Hepatoprotective Effects of Calotropis procera (Ait.) R. Br Root Bark Extracts against Diethylnitrosamine Induced Liver Injury in Rats

Background: Diethylnitrosamine (DEN) is a hepatotoxin whose metabolic activation by liver cytochromes P450 is responsible for the necrosis, mutagenicity and carcinogenicity of liver cells. The purpose of this study was to evaluate the protective effects of Calotropis procera roots bark against DEN induced hepatocellular damage in rats.
 Material and Methods: Hepatoprotective activity of the ethanolic extract of Calotropis procera root bark were evaluated by induction of liver injury with DEN in Wistar male rats distributed in six groups of six. Serum hepatic markers, alanine amino transferase (ALAT), aspartate amino transferase (ASAT), alkaline phosphatase (ALP), total protein and albumin were evaluated and the enzymes antioxidant activities, superoxide dismutase (SOD) and catalase, as well as the level of malonedialdehyde (MDA) were determined in the liver homogenate. Histological analysis was carried out on sections of rat livers. Phytoconstituents have also been studied. Results: Pretreatment of rats with the extract showed a significant decrease in ALAT, ASAT and ALP while there was an increase in total protein and albumin compared to rats treated only with DEN. It also showed a significant increase in SOD and catalase and a decrease in MDA levels suggesting the hepatoprotective effect of the extract. Observation of liver sections confirmed the results of the biochemical parameters which would attest that the extract is hepatoprotective. Phytoconstituents such as sterols, triterpenes and phenolic compounds have been demonstrated.
 Conclusion: Ethanolic extract of Calotropis procera roots bark has shown hepatoprotective effects that could be due to its content in sterols and triterpenic and phenolic compounds.

In situ detection of fatty acid C=C positional isomers by coupling on-tissue mCPBA epoxidation with infrared matrix-assisted laser desorption electrospray ionization mass spectrometry.

Unsaturated fatty acids (UFAs) play vital roles in regulating cellular functions. In-depth structural characterization of UFAs such as localizing carbon-carbon double bonds is fundamentally important but poses considerable challenges in mass spectrometry (MS) given that the most widely accessible ion activation method, low-energy collision-induced dissociation (CID), primarily generates uninformative fragments (e.g., neutral loss of CO2 ) that are not suggestive of the double-bond positions. m-Chloroperoxybenzoic acid (mCPBA) was uniformly deposited onto the sample slides using a TM Sprayer, converting the carbon-carbon double bonds into epoxides under ambient conditions. The epoxidation product was ionized in situ by infrared matrix-assisted laser desorption electrospray ionization mass spectrometry (IR-MALDESI-MS), and subsequently cleaved via CID, generating a diagnostic ion pair associated with the double-bond position. The reaction efficiency, sensitivity and relative quantification capability of the method were validated with five UFA standards dried on glass slides, and then this strategy was demonstrated on thin tissue sections of rat liver and human bladder. The mCPBA reaction yielded conversion rates in the range of 44-60% in 10min with high specificity and sensitivity. Further tandem mass spectrometry (MS/MS) of the mono-epoxidized products generated informative fragment ions specific to the double-bond positions, and relative quantification of positional isomers in binary mixtures was performed across a wide mole fraction from 0 to 1. An innovative spiral scan pattern was utilized during data acquisition, elucidating the major isomeric compositions of multiple UFAs from a tissue section in a single run. The on-tissue mCPBA epoxidation was implemented into an ambient MS imaging workflow to offer a rapid and simple way for in situ identification and relative quantification of double-bond positional isomers without the requirement for instrument modification. The method can be readily implemented on many other MS platforms to reveal the role of double-bond positional isomers in lipid biology and to discover potential biomarkers.

Subacute safety assessment of recombinant Lactococcus lactis on the gut microbiota of male Sprague-Dawley rats.

Lactococcus lactis strain pGSMT/MG1363 is a genetically modified microorganism (GMM) that constitutively expresses human metallothionein-I fusion protein to combine with intracellular lead. Unlike traditional probiotics, pGSMT/MG1363 lacks a history of safe use in food. Administration of microorganism could influence the gut microbial community and consequently confer health benefits or cause disadvantages to the host. To date, little has been done to assess the influence of recombinant strain pGSMT/MG1363 on the stability of gut microbiota. Liver, testis and kidney sections of male Sprague-Dawley rats orally administered pGSMT/MG1363 for 6 weeks showed normal structure and no pathological damage. There were no adverse effects on the analyzed serum biochemical parameters between the pGSMT/MG1363 group and the MG1363 group. Principal coordinate analysis showed that, compared with the MG1363 group, the 6-week-old fecal gut microbiota of rats fed with pGSMT/MG1363 was not significantly different (Adonis, P=0.802). pGSMT/MG1363 treatment for 6 weeks did not significantly change the relative abundance of gut microbiota at the phylum and genus levels in comparison with MG1363 treatment. Compared to the non-GM strain MG1363 group, administration of the recombinant strain pGSMT/MG1363 for 6 weeks showed no adverse effects on the analyzed physiological parameters and gut microbial compositions of male Sprague-Dawley rats. The results suggested that, in terms of gut microbiota stability, pGSMT/MG1363 could be considered as safe as MG1363, at least for short-term intake. Further toxicological evaluations still need to be considered before drawing a definite conclusion concerning the safe use of pGSMT/MG1363. © 2021 Society of Chemical Industry.

Metal Induced Risk of Diabetes Mellitus Due to Toxicological Effects of Mercury: Influence of Environmental Threats

Diabetes Mellitus (DM) is a condition of hyperglycemia due to defects of insulin secretion and/or insulin action. Toxic metals such as lead, nickel, cadmium, arsenic and mercury have been identified which accumulate in various biological samples from T2D (type 2 diabetes) patients through environmental pollution and food chain. Present study will elucidate the toxicological effects of mercury (II) chloride in the pancreatic islets and liver tissues of rat which leads to dysfunction and degeneration of pancreatic islets and liver. Photomicrograph of histology of treated pancreas exhibited the disruption of islets, disorientation of cells and disruption of connective tissue septa. In mercury (II) chloride treated group pancreatic cells were found to be pyknotic and cellular death was confirmed by membrane rupture and necrosis. Alteration of blood glucose levels were observed by glucose tolerance test. The liver sections of rats treated with mercury (II) chloride showed modification in the structure of this organ. Treated liver showed lower periodic acid/Schiff response. In this study, changes in the architecture of pancreatic islets as well as liver may be the reason behind diabetes.

High subzero cryofixation: A technique for observing ice within tissues

While various fixation techniques for observing ice within tissues stored at high sub-zero temperatures currently exist, these techniques require either different fixative solution compositions when assessing different storage temperatures or alteration of the sample temperature to enable alcohol-water substitution. Therefore, high-subzero cryofixation (HSC), was developed to facilitate fixation at any temperature above −80 °C without sample temperature alteration. Rat liver sections (1 cm2) were frozen at a rate of −1 °C/min to −20 °C, stored for 1 h at −20 °C, and processed using classical freeze-substitution (FS) or HSC. FS samples were plunged in liquid nitrogen and held for 1 h before transfer to −80 °C methanol. After 1, 3, or 5 days of −80 °C storage, samples were placed in 3% glutaraldehyde on dry ice and allowed to sublimate. HSC samples were stored in HSC fixative at −20 °C for 1, 3, or 5 days prior to transfer to 4 °C. Tissue sections were paraffin embedded, sliced, and stained prior to quantification of ice size. HSC fixative permeation was linear with time and could be mathematically modelled to determine duration of fixation required for a given tissue depth. Ice grain size within the inner regions of 5 d samples was consistent between HSC and FS processing (p = 0.76); however, FS processing resulted in greater ice grains in the outer region of tissue. This differed significantly from HSC outer regions (p = 0.016) and FS inner regions (p = 0.038). No difference in ice size was observed between HSC inner and outer regions (p = 0.42). This work demonstrates that HSC can be utilized to observe ice formed within liver tissue stored at −20 °C. Unlike isothermal freeze fixation and freeze substitution alternatives, the low melting point of the HSC fixative enables its use at a variety of temperatures without alteration of sample temperature or fixative composition.

18F]Atorvastatin: synthesis of a potential molecular imaging tool for the assessment of statin-related mechanisms of action

BackgroundStatins are lipid-lowering agents that inhibit cholesterol synthesis and are clinically used in the primary and secondary prevention of cardiovascular diseases. However, a considerable group of patients does not respond to statin treatment, and the reason for this is still not completely understood. [18F]Atorvastatin, the 18F-labeled version of one of the most widely prescribed statins, may be a useful tool for statin-related research.Results[18F]Atorvastatin was synthesized via an optimized ruthenium-mediated late-stage 18F-deoxyfluorination. The defluoro-hydroxy precursor was produced via Paal-Knorr pyrrole synthesis and was followed by coordination of the phenol to a ruthenium complex, affording the labeling precursor in approximately 10% overall yield. Optimization and automation of the labeling procedure reliably yielded an injectable solution of [18F]atorvastatin in 19% ± 6% (d.c.) with a molar activity of 65 ± 32 GBq·μmol−1. Incubation of [18F]atorvastatin in human serum did not lead to decomposition. Furthermore, we have shown the ability of [18F]atorvastatin to cross the hepatic cell membrane to the cytosolic and microsomal fractions where HMG-CoA reductase is known to be highly expressed. Blocking assays using rat liver sections confirmed the specific binding to HMG-CoA reductase. Autoradiography on rat aorta stimulated to develop atherosclerotic plaques revealed that [18F]atorvastatin significantly accumulates in this tissue when compared to the healthy model.ConclusionsThe improved ruthenium-mediated 18F-deoxyfluorination procedure overcomes previous hurdles such as the addition of salt additives, the drying steps, or the use of different solvent mixtures at different phases of the process, which increases its practical use, and may allow faster translation to clinical settings. Based on tissue uptake evaluations, [18F]atorvastatin showed the potential to be used as a tool for the understanding of the mechanism of action of statins. Further knowledge of the in vivo biodistribution of [18F]atorvastatin may help to better understand the origin of off-target effects and potentially allow to distinguish between statin-resistant and non-resistant patients.

Hepatoprotective and hematological effects of Justicia secunda Vahl leaves on carbon tetrachloride induced toxicity in rats

ABSTRACTJusticia secunda Vahl is an exotic plant that is used to treat medical problems. We investigated the hepatoprotective and hematological effects of aqueous extracts of J. secunda leaves on carbon tetrachloride induced toxicity in rats. Leaf extracts were prepared using hot and cold extraction methods to obtain a hot extract of J. secunda leaves (JSHAE) and a cold extract of J. secunda leaves (JSCAE). Total phenol and flavonoid measurements and antioxidant assays were performed to determine the extract with the greater antioxidant activity. JSHAE was the more effective extract for treatment of carbon tetrachloride (CCl4) induced hepatotoxicity and hepatotoxicity in rats. Silymarin was used as a standard for comparison. We found that JSHAE contained more total phenol and flavonoid than JSCAE. JSHAE exhibited significantly greater ferric reducing antioxidant power and 1,1-diphenyl-2-picryl hydrazyl and thiobarbituric acid scavenging activity than JSCAE. We also found that in vivo, 100 and 200 mg/kg JSHAE significantly reduced plasma aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase and total bilirubin levels following CCl4 induced toxicity compared to untreated rats. JSHAE treated animals exhibited white blood cell, red blood cell, hemoglobin, hematocrit, platelet and procalcitonin levels that were comparable to control animals. Liver sections of rats treated with 200 mg/kg. JSHAE exhibited no abnormalities.

Localization of ATP-sensitive K+ channel subunits in rat liver.

BACKGROUNDATP-sensitive K+ (KATP) channels were originally found in cardiac myocytes by Noma in 1983. KATP channels were formed by potassium ion-passing pore-forming subunits (Kir6.1, Kir6.2) and regulatory subunits SUR1, SU2A and SUR2B. A number of cells and tissues have been revealed to contain these channels including hepatocytes, but detailed localization of these subunits in different types of liver cells was still uncertain.AIMTo investigate the expression of KATP channel subunits in rat liver and their localization in different cells of the liver.METHODSRabbit anti-rat SUR1 peptide antibody was raised and purified by antigen immunoaffinity column chromatography. Four of Sprague-Dawley rats were used for liver protein extraction for immunoblot analysis, seven of them were used for immunohistochemistry both for the ABC method and immunofluorescence staining. Four of Wistar rats were used for the isolation of hepatic stellate cells (HSCs) and Kupffer cells for both primary culture and immunocytochemistry.RESULTSImmunoblot analysis showed that the five kinds of KATP channel subunits, i.e. Kir6.1, Kir6.2, SUR1, SUR2A, and SUR2B, were detected in liver. Immunohistochemical staining showed that Kir6.1 and Kir6.2 were weakly to moderately expressed in parenchymal cells and sinusoidal lining cells, while SUR1, SUR2A, and SUR2B were mainly localized to sinusoidal lining cells, such as HSCs, Kupffer cells, and sinusoidal endothelial cells. Immunoreactivity for SUR2A and SUR2B was expressed in the hepatocyte membrane. Double immunofluorescence staining further showed that the pore-forming subunits Kir6.1 and/or Kir6.2 colocalized with GFAP in rat liver sections and primary cultured HSCs. These KATP channel subunits also colocalized with CD68 in liver sections and primary cultured Kupffer cells. The SUR subunits colocalized with GFAP in liver sections and colocalized with CD68 both in liver sections and primary cultured Kupffer cells. In addition, five KATP channel subunits colocalized with SE-1 in sinusoidal endothelial cells.CONCLUSIONObservations from the present study indicated that KATP channel subunits expressed in rat liver and the diversity of KATP channel subunit composition might form different types of KATP channels. This is applicable to hepatocytes, HSCs, various types of Kupffer cells and sinusoidal endothelial cells.

Hepatoprotective and Antioxidant Activities of Justicia gendarussa Leaf Extract in Carbofuran-Induced Hepatic Damage in Rats.

In folk medicines, Justicia gendarussa (J. gendarussa) is used as a depurative herb for treating fever, pain, and cancer and as laxative for constipation. The aim of the present investigation was to evaluate the hepatoprotective effect of the leaf methanol extract of J. gendarussa leaf (J gMe) against carbofuran (CF)-intoxicated liver injuries in Sprague-Dawley rats, along with the antioxidant activity of this extract. For this purpose, levels of serum diagnostic markers, hepatic antioxidant enzymes, and liver histo-architecture were employed to justify the protective efficacy of J gMe. In addition, the phenolic and flavonoid contents of the extract were quantified, and antioxidant activity was investigated using the 1,1-diphenyl-2-picrylhydrazyl (DPPH), nitric oxide, hydrogen peroxide, and hydroxyl free radical scavenging assays. Results revealed that the leaf extract caused a significant (<0.05, <0.01) decrease of the level of hepatic enzymes, triglycerides, and bilirubin and an increase of the total protein. J gMe has also significantly (<0.05, <0.01) lowered the level of malonylaldehyde. Carbofuran markedly suppressed hepatic antioxidant enzymes, however, the leaf extract significantly augmented these enzymes. The hepatoprotective effect was demonstrated by the improvement in the histo-architectural features of liver sections of CF-intoxicated rats treated with J gMe at 500 mg/kg dose. In addition, J gMe showed moderate total phenolic and total flavonoid content, whereas the IC50 values of DPPH, nitric oxide, hydrogen peroxide, and hydroxyl free radical scavenging assays were 71.31 ± 0.42, 134.82 ± 0.14, 47.69 ± 0.38, and 118.44 ± 0.30 μg/mL, respectively. In conclusion, the present study suggests the protective role of J gMe against hepatic injury induced by CF, which may be attributed to its higher antioxidant properties and thereby scientifically justifies its traditional use.

Targeting AngII/AT1R signaling pathway by perindopril inhibits ongoing liver fibrosis in rat.

The renin-angiotensin system (RAS) has a substantial role in liver fibrosis, cirrhosis, and portal hypertension. Hence, targeting RAS through angiotensin-converting enzyme (ACE) inhibitors can mend hepatic fibrosis; the current study was designed to examine the potential fibrosis inhibition activity of perindopril using a rat model of liver fibrosis induced by thioacetamide (TAA). Four groups of rats were used throughout this study, Group I (control group); rats received the vehicle. TAA was used for inducing liver fibrosis in rats by intraperitoneal injection of 200-mg/kg body weight twice a week for 6 weeks. Group II served as (TAA group). Rats of Groups III and IV were given perindopril at doses of 2 and 8 mg/kg 2 weeks after TAA administration and continued concomitantly with TAA till the end of the experiment. Injection of TAA resulted in a significant increase in aminotransferases' activities and bilirubin with a significant decrease in serum albumin and total protein and a significant decrease in hepatic content of GSH and SOD. Additionally, TAA injection raised the hepatic content of TGF-β1, α-SMA, TNF-α, and level of MDA. Histological and immunohistochemical data presented marked fibrosis in liver sections of TAA-administrated rats with increased collagen deposition, elevated METAVIR scoring, and increased expression of α-SMA, caspase-3, and AT1R. Oral dosing of perindopril for 4 weeks concomitant with TAA could mend the altered parameters near to normal values and abolished the ongoing fibrosis extension. In conclusion, these results demonstrated that perindopril, as ACE inhibitor, could grant a superior remedial nominee in preventing liver fibrosis progression through targeting angiotensin II formation.

Hepatotoxic Effects of Microcystis aeruginosa (PCC 7820) on Wister Rats

The potential toxic effect of Microcystis aeruginosa through oral contamination is becoming important. The present study was carried out to find the possible hepatotoxic effects of toxic M. aeruginosa (PCC 7820) on male Wistar rats as animal model. Hepatotoxicity assessment 1vas done by estimation of serum hepatic enzyme levels of r-Glutamyl transferase (GGT). Alkaline phosphatase (ALP). Alanine aminotransferase (ALT/GPT), Aspartate aminotransferese (GOT/AST). Rat treated with non toxic (CYA -/3) and toxic non homogenized M. aeruginosa (PCC 7820) were normal in appearance where as rats receiving homogenized toxic M aeruginosa (PCC 7820) were lethargic and gained less weight which indicates a possible toxic effect on the test animals. The absolute and relative (% body weight) mean weight of liver and kidneys ofthe homogenous toxic M aeruginosa (PCC 7820) treated animal lj'ere lower than those who received non homogenized toxic M aeruginosa (PCC 7820) and the difference is statistically significant (p&lt;0. 00J). Liver sections of rats receiving fresh toxic M aeruginosa and the homogenized toxic M aeruginosa did not show lymphatic infiltration or signs of necrosis. Rats treated 'with homogenized M aeruginosa showed lowering of absolute liver weight compared to the fresh M aeruginosa treated rats. Statisticcally significant (p&lt;0.005) increase levels of serum y-Glutamyl transferase (GGT) was detected in rats 14 days after oral administration with homogenized toxic M aeruginosa (PCC 7820). There was no significant differentfound in serum Alanine aminotransferase (ALT/GPT) concentration between treated and control groups. The results indicate that prolonged oral administration of homogenized M. aeruginosa lead to functional hepatotoxicity in male Wister rats without evident histopathological liver necrosis.

Effects of Moringa oleifera on oxidative stress, apoptotic and inflammatory biomarkers in streptozotocin-induced diabetic animal model

Aim of the studyMoringa oleifera is a highly nutrious medicinal plant with various potential applications in the treatment of diabetes, oxidative stress, bacterial infections and inflammation. This study assessed the effects of the leaf extract of Moringa oleifera on the expression of specific apoptotic and anti-inflammatory biomarkers in the liver and kidney of streptozotocin (STZ)-induced diabetes in male Wistar rats. Materials and methodsRats weighing between 200 and 250 g were randomly divided into four groups: non-diabetic control (NC), non-diabetic Moringa treated (NC + MO), diabetic control (DM) and diabetic Moringa treated (DM + MO). Diabetes was induced in rats by a single intraperitoneal injection of STZ (55 mg/kg) and treated with 250 mg/kg of MO extracts. Interleukin (IL-1α), interleukin (IL-12) and interleukin (IL-18) were assayed in the kidney and liver homogenates. Kidney and liver sections of rats were assessed for apoptotic cell death markers. Antioxidant capacity and phytochemical contents of methanol extract of MO were estimated. ResultsMO extract shows the presence of polyphenols, flavonols and alkaloids. Diabetic rats treated with MO was compared with untreated diabetic rats. A significant decrease in serum NFkβ levels was observed. There was upregulation of BCL-2 in both the kidney and liver in diabetic treated rats. A reduction in interleukin levels in the kidney (IL-18) and liver (IL-1α, IL-18) interleukin levels were observed in diabetic treated rats. ConclusionThis study demonstrates that MO exerted anti-apoptotic and anti-inflammatory properties in diabetic rats due to its capacity to reduce nephrotoxic and hepatotoxic damage induced by STZ and to regulate the expression and suppression of specific apoptotic cell death markers. The high phytochemical and antioxidant content of MO leaves may be responsible for its anti-apoptotic and anti-inflammatory effects.

Protective Effect of Curcuma Against Chromium Hepatotoxicity in Rats

This study was carried out to investigate the antioxidant effects of curcuma against chromium-induced alterations in hepatic indices and dysfunctions in the antioxidant system. Forty maleWistarrats were randomly divided into four groups and were treated for 30 consecutive days. The control group (0-0) received per os mineral water and normal diet. The second group (0-Cur) received mineral water and an experimental diet containing 2% of curcuma powder, whereas the third group (Cr-0) was orally fed (per os) with 15 mg/kg body weight/day of potassium dichromate and normal diet. The last group (Cr-Cur) received per os 15 mg/kg of potassium dichromate and a diet with 2% of curcuma. The treatment by chromium was found to elicit a perturbation in biochemical parameters producing a significant increase in glycemia, triglycerides, cholesterol, ALP, ALT, AST, and LDH levels. On the contrary, a significant reduction was observed in the oxidative stress-related parameters (GSH, GPx, CAT, and GST). Moreover, we noticed that liver sections of rats intoxicated with chromium showed a disrupted architecture. However, the administration of curcuma revealed an intense reduction in the oxidative stress induced by chromium, ameliorating the levels of the majority of the previous parameters. The data of this study revealed the potent antioxidant effects of curcuma in reducing oxidative stress damage induced by the hexavalent chromium.