Portal Area Research Articles

Mesenchymal Stem Cells in Rat Hepatic Fibrosis MModel: Benefits? Mechanism of Action?: 2017 Presidential Poster Award

Introduction: Hepatic fibrosis may evolve to liver cirrhosis which may ultimately develop into hepatocellular carcinoma. Mesenchymal stem cells (MSCs) have potential to trans-differentiate into hepatic cells and thus may be a curative option in liver cirrhosis. Natural killer (NK) cells are a component of the innate immune system that destroy compromised host cells and have critical antifibrotic role. Aim: To show the effects of bone marrow MSCs transferred via the tail vein to the rats having hepatic fibrosis produced by common bile duct ligation (CBDL) with a special focus on the changes of the NK population distribution. Methods: Rats were divided into three groups; 1- CBDL rats that were injected with phosphate-buffered saline (PBS), 2- CBDL rats injected with MSCs, 3- healthy rats that were sham operated and given MSCs. All rats were sacrificed at the end of the experiment. We analyzed the in vivo functional outcomes by measuring the serum glucose, bilirubin and ALT levels. Splenocytes isolated from the collected spleens were cultured and the supernatants were analyzed for IL (interleukin)-1α, TNF (tumor necrosis factor)-α, IFN (interferon)-γ and IL-10 using flow cytometry. Results: Liver fibrosis developed in CBDL rats while the healthy rats did not show any alteration in liver architecture. Although healthy group had significantly lower serum bilirubin, ALT and glucose levels than CBDL rats, the difference regarding those parameters did not differ between groups 1 and 2. T-lymphocyte proliferation was significantly suppressed in group 2 compared to group 1. IL-1α, TNF-α and IFN-γ levels in supernatants of cultured lymphocytes in group 2 were significantly higher than group 3 while they tended to be lower than group 1. NK and Treg cells in peripheral blood significantly increased in group 2 compared to group 1. Histologically, hepatic fibrosis scores were significantly alleviated in group 2 compared to group 1 (p=0.039). NK cells in the general hepatic parenchyma and in the portal areas were both increased significantly in group 2 compared to group 1. Conclusion: Our findings suggest bone marrow derived MSCs may be beneficial in alleviating the hepatic fibrosis by suppressing the T cell proliferation and the proinflammatory cytokines and inducing NK cell recruitment to liver parenchyma in rats. Thus, MSC treatment may appear promising in liver injury and future clinical therapies are warranted. Immunophenotyping of leukocytes. CD3+CD4+ T lymphocyte, CD3+CD8+ Cytotoxic T cell, CD4+CD25+ T regulatory cell and CD161a NK cell in all groups by flow cytometry. (B) CD3+CD4+ % T lymphocyte, CD3+CD8+ % Cytotoxic T cell, CD4+CD25+ %T regulatory cell and CD161a NK cells were displayed statistically in all groups..*P < 0.05, **P < 0.01.Figure

Portal Vein Dopplerflowmetry in healthy sheep according to age

ABSTRACT: Pulsed Doppler ultrasound was used to evaluate portal blood flow, portal velocity and portal congestion index in 24 healthy sheep divided into groups (lambs, yearlings and ewes), according to age. Measurements were performed at the 11th right intercostal space using ideal insonation angle and uniform insonation method. Mean values obtained in each group were compared with one-way ANOVA, followed by Tukey post-hoc test. Portal velocity and portal blood flow were statistically similar between the groups (P&gt;0.05). Mean portal velocity were 17.75; 17.13 and 16.75; while mean portal blood flow were 26.65; 31.04 and 24.32 for lambs, yearlings and ewes, respectively. Portal congestion index was statistically distinct between the groups and values for lambs, yearlings and ewes were 0.009; 0.058 and 0.09, respectively (P&lt;0.01). Statistical differences were observed in portal vein diameter, portal vein area and portal congestion index between the groups, presumably due to influence of weight and not to age.

The protective effect of vitamin D against carbon tetrachloride damage to the rat liver

We investigated the protective effect of vitamin D against liver damage caused by carbon tetrachloride (CCl4). Twenty-four male rats were divided into four equal groups: G1, untreated controls; G2, administered CCl4; G3, administered both CCl4 and vitamin D for 10 weeks; G4, administered CCl4 for 10 weeks and vitamin D for 12 weeks. At the end of experiment, intracardiac blood samples were taken and liver samples were removed. Hepatic damage due to CCl4 was assessed using biochemistry and histopathology. Glutathione (GSH) levels decreased, while malondialdehyde (MDA) levels increased in liver tissues of G2. Alanine transaminase (ALT), alkaline phosphatase (ALP), and gamma-glutamyl-transaminase (GGT) levels increased, while albumin (ALB) levels decreased. Hepatocyte degeneration, lobular disorder, sinusoid dilation, focal necrotic areas, hyperemia, and glycogen loss were observed. Hepatic fibrosis was observed around portal areas and central veins. Bridging fibrous septa were formed between portal veins. By immunohistochemistry, both matrix metalloproteinase-9 (MMP-9) and desmin reactivity were increased. All aspects of liver damage were at least partially prevented in rats treated with vitamin D. Vitamin D appears to act as an antioxidant and anti-fibrotic to protect the rat liver against damage.

Energy restriction affect liver development in Hu sheep ram lambs through Hippo signaling pathway.

This study aimed to evaluate the effects of dietary energy restriction on postnatal liver development in Hu sheep ram lambs. A total of 16 ram lambs were randomly divided into two groups: 100% energy requirements diet and 55% energy requirements diet, which were fed for 75 d. Results showed that the final body and liver weights decreased with energy restriction (p <0.05). Energy restriction caused a significant decrease in the levels of circulating insulin-like growth factor 1 (IGF1) and an increase in growth hormone secretion (p <0.05), which can be explained by the decreased mRNA expression levels of the growth hormone receptor (GHR) and IGF1 (p <0.05). The proliferating cell nuclear antigen (PCNA), Ki-67 and apoptosis-related proteins (BAX and BCL2) were mainly located in the nucleus and the cytoplasm of hepatocytes, respectively. The transcription and protein levels of PCNA and BAX were significantly decreased and increased by energy restriction, respectively (p <0.05). The caspase9 and caspase3 mRNA and activity were increased in energy restriction group (p <0.05). Moreover, Hippo signaling pathway proteins [mammalian sterile 20-like protein kinase 1 (MST1), large tumor suppressor kinase 1 (LATS1), and yes-associated protein 1 (YAP1)] were mainly observed around the hepatic portal area, and the expression levels of their mRNA and proteins were significantly decreased in energy restriction group (p <0.05). In summary, energy restriction in ram lambs impairs liver development by increasing apoptosis, which may occur via the Hippo signaling pathway.

Gamma-irradiated β-glucan modulates signaling molecular targets of hepatocellular carcinoma in rats.

β-glucans are one of the most abundant forms of polysaccharides known as biological response modifiers which influence host's biological response and stimulate immune system. Accordingly, this study was initiated to evaluate irradiated β-glucan as a modulator for cellular signaling growth factors involved in the pathogenesis of hepatocellular carcinoma in rats. Hepatocellular carcinoma was induced with 20 mg diethylnitrosamine/kg BW. Rats received daily by gastric gavage 65 mg irradiated β-glucan/kg BW. It was found that treatment of rats with diethylnitrosamine induced hepatic injury and caused significant increase in liver injury markers with a concomitant significant increase in both hepatic oxidative and inflammatory indices: alpha-fetoprotein, interferon gamma, and interleukin 6 in comparison with normal and irradiated β-glucan-treated groups. Western immunoblotting showed a significant increase in the signaling growth factors: extracellular signal-regulated kinase 1 and phosphoinositide 3-kinase proteins in a diethylnitrosamine-treated group while both preventive and therapeutic irradiated β-glucan treatments recorded significant improvement versus diethylnitrosamine group via the modulation of growth factors that encounters hepatic toxicity. The transcript levels of vascular endothelial growth factor A and inducible nitric oxide synthase genes were significantly higher in the diethylnitrosamine-treated group in comparison with controls. Preventive and therapeutic treatments with irradiated β-glucan demonstrated that the transcript level of these genes was significantly decreased which demonstrates the protective effect of β-glucan. Histological investigations revealed that diethylnitrosamine treatment affects the hepatic architecture throughout the significant severe appearance of inflammatory cell infiltration in the portal area and congestion in the portal vein in association with severe degeneration and dysplasia in hepatocytes all over hepatic parenchyma. The severity of hepatic architecture changes was significantly decreased with both β-glucan therapeutic and preventive treatments. In conclusion, irradiated β-glucan modulated signal growth factors, vascular endothelial growth factor A, extracellular signal-regulated kinase 1, and phosphatidylinositol-3-kinase, which contributed to experimental hepatocarcinogenesis.

Hepatic iron overload is associated with hepatocyte apoptosis during Clonorchis sinensis infection

BackgroundHepatic iron overload has been implicated in many liver diseases; however, whether it is involved in clonorchiasis remains unknown. The purpose of this study is to investigate whether Clonorchis sinensis (C. sinensis) infection causes hepatic iron overload, analyze the relationship between the iron overload and associated cell apoptosis, so as to determine the role of excess iron plays in C. sinensis-induced liver injury.MethodsThe Perls’ Prussian staining and atomic absorption spectrometry methods were used to investigate the iron overload in hepatic sections of wistar rats and patients infected with C. sinensis. The hepatic apoptosis was detected by transferase uridyl nick end labeling (TUNEL) methods. Spearman analysis was used for determining the correlation of the histological hepatic iron index and the apoptotic index.ResultsBlue iron particles were deposited mainly in the hepatocytes, Kupffer cells and endothelial cells, around the liver portal and central vein area of both patients and rats. The total iron score was found to be higher in the infected groups than the respective control from 8 weeks. The hepatic iron concentration was also significantly higher in treatment groups than in control rats from 8 weeks. The hepatocyte apoptosis was found to be significantly higher in the portal area of the liver tissue and around the central vein. However, spearman’s rank correlation coefficient revealed that there was a mildly negative correlation between the iron index and hepatocyte apoptosis.ConclusionsThis present study confirmed that hepatic iron overload was found during C. sinensis infection. This suggests that iron overload may be associated with hepatocyte apoptosis and involved in liver injury during C. sinensis infection. Further studies are needed to investigate the molecular mechanism involved here.

The evaluation of non‐invasive multi‐slice spiral computed tomography‐based indices for the diagnosis and prognosis prediction of liver cirrhosis

This study aimed to evaluate easily available computed tomography (CT)-based parameters for assessing the presence and severity of cirrhosis and predicting complications in Chinese patients with cirrhosis. CT-based morphological indices were determined in 167 patients with cirrhosis and 244 healthy volunteers. The correlation of morphological indices with Child-Pugh score and cirrhotic complications was analyzed using Spearman's correlation analysis. The area under the receiver operating characteristic curve (AUROC) was used to analyze the diagnostic performance of the indices. Sensitivity and specificity were determined. Patients with cirrhosis had a lower total liver volume (TLV) and a larger total splenic volume (SV) than healthy individuals. There was a significant difference in the portal venous diameter, splenic venous diameter and portal venous cross-sectional area between the two groups. A low TLV/SV ratio was strongly associated with liver cirrhosis; with a cut-off value of 4.27 for the diagnosis of cirrhosis TLV/SV had a sensitivity of 87.7% and a specificity of 84.9%, and AUROC of 0.921. Further analysis showed that TLV/SV was accurate in discriminating between mild and moderate/severe cirrhosis and could be used for predicting complications of cirrhosis. The easily available parameters of CT can accurately evaluate the severity of cirrhosis in Chinese patients.

Cholangiocytes: No Longer Cinderellas to the Hepatic Regenerative Response

Biliary ductal cells proliferate from the portal areas of chronically damaged livers, but their significance to regeneration has been controversial. A recent article in Nature by Raven etal. (2017) now shows that blocking hepatocyte replication is essential for the hepatic differentiation of ductular cells after liver damage.

Systemic effects of H2S inhalation at human equivalent dose of pathologic halitosis on rats

Objectives: Halitosis is composed by hundreds of toxic gases. It is still not clear whether halitosis gases self-inhaled by halitosis patients cause side effects. The aim of the study was to investigate the effect of H2S inhalation at a low concentration (human equivalent dose of pathologic halitosis) on rats.Materials and methods: The threshold level of pathologic halitosis perceived by humans at 250 ppb of H2S was converted to rat equivalent concentration (4.15 ppm). In the experimental group, 8 rats were exposed to H2S via continuous inhalation but not the control rats. After 50 days, blood parameters were measured and tissue samples were obtained from the brain, kidney and liver and examined histopathologically to determine any systemic effect.Results: While aspartate transaminase, creatine kinase-MB and lactate dehydrogenase levels were found to be significantly elevated, carbondioxide and alkaline phosphatase were decreased in experimental rats. Other blood parameters were not changed significantly. Experimental rats lost weight and became anxious.Histopathological examination showed mononuclear inflammatory cell invasion in the portal areas, nuclear glycogen vacuoles in the parenchymal area, single-cell necrosis in a few foci, clear expansion in the central hepatic vein and sinusoids, hyperplasia in Kupffer cells and potential fibrous tissue expansion in the portal areas in the experimental rats. However, no considerable histologic damage was observed in the brain and kidney specimens.Conclusions: It can be concluded that H2S inhalation equivalent to pathologic halitosis producing level in humans may lead to systemic effects, particularly heart or liver damage in rats.

Morphological and histological study of the liver in migratory starling bird (Sturnus vulgaris)

The present work was aimed to form the baseline data of normal morphological and histological structure features of liver in migratory starling (Sturnus vulgaris). Anatomically, the starling liver bird was dark red -brown in colour and located in the cranial third of the abdominal cavity and consisted of undivided lobes (left and right). The liver right lobe was larger than the left. Histological examination revealed that the liver parenchyma was covered by a connective tissue capsule which appears to be thicker in the rim of liver lobes than other area in the liver lobe. Liver parenchyma was arranged in an unlimited hepatic lobules, which composed of polygonal hepatocytes organized as irregular, radial interconnecting cords or laminae of one or two cells thickness around a central vein and separated by blood sinusoids. In the boundary of each lobule showed a portal area which consists of a branch of hepatic artery; one or more branches of hepatic vein and one to four branches of the bile duct which lining by cuboidal cells that characterized by their empty non-staining cytoplasm. The histochemical observation by using PAS staining in the current study revealed that the glycogen granules arranged close to the central vein and in the rim of liver lobules.The present work was aimed to form the baseline data of normal morphological and histological structure features of liver in migratory starling (Sturnus vulgaris). Anatomically, the starling liver bird was dark red -brown in colour and located in the cranial third of the abdominal cavity and consisted of undivided lobes (left and right). The liver right lobe was larger than the left. Histological examination revealed that the liver parenchyma was covered by a connective tissue capsule which appears to be thicker in the rim of liver lobes than other area in the liver lobe. Liver parenchyma was arranged in an unlimited hepatic lobules, which composed of polygonal hepatocytes organized as irregular, radial interconnecting cords or laminae of one or two cells thickness around a central vein and separated by blood sinusoids. In the boundary of each lobule showed a portal area which consists of a branch of hepatic artery; one or more branches of hepatic vein and one to four branches of the bile duct which lining by cuboidal cells that characterized by their empty non-staining cytoplasm. The histochemical observation by using PAS staining in the current study revealed that the glycogen granules arranged close to the central vein and in the rim of liver lobules. Keywords: Birds, Liver morphology, liver histology.

Effect of Psoralea corylifolia in treating fatty liver disease in juvenal mouse by inhibiting hepatic NF-κB activation

To investigate the mechanism and effect of Psoralea corylifolia(PC) in the treatment of NAFLD in juvenal mice. The NAFLD model in juvenal mice was established by feeding high-fat diet. Then PC herbal granules (at low and high dose) were administered for 5 weeks. Blood glucose (FBG, PG-1 h/2 h), blood lipid (TC, TG, HDL-C, LDL-C), fasting insulin, liver function (ALT, AST) were examined. HOMA-IR was calculated. Hepatic histological changes were observed. The content of TG, inflammatory factor (TNF-α, IL-8) and protein expressions of CD44, NF-κB p65, p-NF-κB p65 in hepatic tissues were determined. The ratio of p-NF-κB p65 to NF-κB p65 (p-p65/p65) was calculated. The result showed that compared with the model group, both PC treatment groups showed reduction in hepatic steatosis, inflammatory cell infiltration and fibroplasia in portal area. HOMA-IR, ALT, AST, FBG, PG-2 h, TC, TG, LDL-C concentrations and hepatic TG content were also significantly decreased, with the reduction of TNF-α, IL-8 contents, CD44 expression and p-p65/p65 ratio in hepatic tissues (P<0.01). High-dose PC group had a better effect than low-dose group (P<0.01, P<0.05). In conclusion, PC is effective in treating hepatic injury, glucolipid metabolism disturbances and fibrosis in juvenal NAFLD mice. The mechanism may be related to inhibition of inflammation and down-regulation of the activation of hepatic NF-κB.

Pathological study of liver lesions in cattle slaughtered at Kirkuk province abattoir

Current study aimed to identify the gross and microscopic lesions and their percentage in liver of cattle slaughtered in Kirkuk province abattoir. A total of 6211 liver sample were examined during period from 01/08/2016 until 01/12/2016 with total cattle liver that showed gross lesions 738 cases. A gross examination was applied to affected liver and tissue sample were taken from lesions for histopathological examination. The result of current study showed that the total liver lesions in cattle is about 11.88% (738/6211) which distributed as liver fluke infestation 3.34% (213/6211), hydatid cyst 3.12% (194/6211), liver abscess 2.29% (142/6211), cholangiohepatitis 2.01% (125/6211), and hepatic hemorrhage with congestion 1.03% (64/6211). The results of gross and microscopic examination of liver infested with liver fluke showed presence of thickening in bile ducts with adult fluke as well as liver atrophy with hyperplasia of epithelial cell in affected bile ducts with dystrophic calcification. While in case of hydatid cyst the lesions composed from presence of larval stage in form of white cyst in different size and their walls composed from hyperplasia of fibrocytes and fibrin strands. In case of hepatic hemorrhage and congestion the result showed sever congestion with staining of liver tissue by blood color, with presence of red blood cells in association with inflammatory cells between hepatocytes. While in case of hepatic abscess we noticed presence of white to yellow solid lesions that randomly distributed in different shape and size composed from necrotic center with infiltration of mononuclear inflammatory cells and hyperplasia of fibrocytes. While in case of cholangiohepatitis the affected area showed paleness with accumulation of bile material inside bile ducts, as well as hyperplasia of affected ducts and fibrosis in portal areas with infiltration of mononuclear inflammatory cells in liver tissue with dystrophic calcification in bile ducts. We concluded from current study that wide spread of hepatic lesions in liver of cattle in Kirkuk province and this result should be taken in more serious action which can result in economic losses as well as possibility of zoonosis of these pathogens to human, also the gross lesions described by current study were identically similar to microscopic lesions.

Oxygen and Glucose as Stimulation Agents for BOLD Functional MR Imaging of Rabbit Liver: A Feasibility Study.

Purpose:To assess the feasibility of using oxygen and glucose as stimulating agents in blood-oxygen-level-dependent (BOLD) Functional Magnetic Resonance Imaging (fMRI) of rabbit liver and analyze the impacts by blood flow.Methods:Pure oxygen inhalation, intravenous injection and oral administration of glucose were given to 11 New Zealand white rabbits to compare the differences of liver , aortic flow (AF), portal vein flow (PVF), aortic area (AA) and portal vein area (PVA) before and at 5 min, 10 min, 20 min, 30 min after administrations. AF and PVF were acquired by two dimensional (2D) Phase Contrast MR (2D-PCMR). The impacts of AF and PVF upon BOLD fMRI were analyzed.Results:AF and PVF declined at 5 min after oxygen inhalation and were significantly different from baseline, then reverted to baseline. No significant difference was observed in liver , AA and PVA before and after oxygen inhalation. AF, PVF, AA and PVA showed no significant difference before and after glucose intravenous injection, while liver increased gradually with significant difference. AF and liver were significantly different before and after glucose oral administration and increased gradually, AA was significantly different before and after glucose administration at 10 min and 20 min. PVF and PVA started to be different from baseline at 10 min. Greatest variation of (19.6%) was induced by glucose oral administration after 30 min.Conclusion:Rabbit liver increasing by glucose intravenous injection is possibly associated with glycogen synthesis, provides the possibility to evaluate liver function. Glucose oral administration demonstrated an optimal comparative effect of raising , however, resulted from the superposition of increased glycogen synthesis and blood flow. Inhalation of pure oxygen didn’t alter the rabbit liver , which may possibly result from an offset between the increased concentration of oxyhemoglobin and decreased blood flow.

N-nitrosamines induced infertility and hepatotoxicity in male rabbits.

N-nitrosamines are widely spread environmental pollutants of well-known toxicity and carcinogenicity in various animal species. These compounds are metabolically activated by cytochrome P450 system predominantly in the liver and in other tissues into more active metabolites leading to generation of both alkylating agents that alkylate DNA and reactive oxygen species. In the current study, we investigated the influence of four types of N-nitrosamines that are commonly present in the environment [methyethylnitrosamine, (MEN), diethylnitrosamine (DEN), diphenylnitroasamine (DPN) and dimethylnitrosamine (DMN)] on both livers and testes of male rabbits through assessment of 17 β-hydroxysteroid dehydrogenase (17 β-HSD) activity. The protein expression of the three cytochrome P450s (CYP11A1, CYP19A1, and CYP21A2) is involved in the steroidogenesis. The levels of testosterone (T) and estradiol (E2) were also determined in the plasma of N-nitrosamines-treated rabbits after one, four-, eight- and twelve weeks of treatment of male New Zealand rabbits with an oral dose of 0.5 mg/kg B.W/day of each compound. In addition, activities of glutathione reductase (GR), superoxide dismutase (SOD), catalase (CAT) and levels of free radicals measured as thiobarbituric acid reactive substances (TBARS), and reduced glutathione (GSH) level were quantified in both livers and testes. The present study showed that levels of free radicals (TBARS) were markedly increased, whereas GSH levels were depleted in the tissues of both livers and testes after treatment of rabbits with any of N-nitrosamines. In addition, all tested N-nitrosamines inhibited the activities of antioxidant enzyme activities (GR, GST, SOD, and CAT) in hepatic and testicular tissues of rabbits after 12 weeks of treatment. Histopathological examination showed that N-nitrosamines caused lymphocytic infiltration with vascular degeneration and necrosis, congestion of central vein with RBCs hemolysis, dilated sinusoids, as well as fibrosis around portal areas were seen in hepatic tissues. In the testes, histopathological examination displayed disorganized seminiferous tubules with degeneration of germinal epithelium and Sertoli cells. Also, spermatogenic cells had pyknotic nuclei and others were detached from basement membranes of seminiferous tubules, edema was seen between seminiferous tubules. Moreover, the present data showed that MEN and DEN down-regulated the protein expression of both CYP19A1 and 21A2 in both livers and testes of male rabbits. In addition, both MEN and DEN decreased levels of testosterone and estradiol in plasma of treated rabbits. On the one hand, DMN and DPN markedly up-regulated the protein expression of CYP19A1 in both hepatic and testicular tissues of treated rabbits. These compounds potentially increased estradiol and decreased testosterone levels. On the other hand, no correlation was found between the expression of CYP11A1 and levels of both testosterone and estradiol. It is concluded that most of tested N-nitrosamines induce different changes, which could be a new mechanism of infertility due to exposure to N-nitrosamines from different environmental sources.

Infection with Opisthorchis felineus induces intraepithelial neoplasia of the biliary tract in a rodent model.

The liver fluke Opisthorchis felineus is a member of the triad of epidemiologically relevant species of the trematode family Opisthorchiidae, and the causative agent of opisthorchiasis felinea over an extensive range that spans regions of Eurasia. The International Agency for Research on Cancer classifies the infection with the liver flukes Opisthorchis viverrini and Clonorchis sinensis as group 1 agents and a major risk factor for cholangiocarcinoma. However, the carcinogenic potential of the infection with O. felineus is less clear. Here, we present findings that support the inclusion of O. felineus in the Group 1 list of biological carcinogens. Two discrete lines of evidence support the notion that infection with this liver fluke is carcinogenic. First, novel oxysterol-like metabolites detected by liquid chromatography-mass spectroscopy in the egg and adult developmental stages of O. felineus, and in bile, sera, and urine of liver fluke-infected hamsters exhibited marked similarity to oxysterol-like molecules known from O. viverrini. Numerous oxysterols and related DNA-adducts detected in the liver fluke eggs and in bile from infected hamsters suggested that infection-associated oxysterols induced chromosomal lesions in host cells. Second, histological analysis of liver sections from hamsters infected with O. felineus confirmed portal area enlargement, inflammation with severe periductal fibrosis and changes in the epithelium of the biliary tract characterized as biliary intraepithelial neoplasia, BilIN. The consonance of these biochemical and histopathological changes revealed that O. felineus infection in this rodent model induced precancerous lesions conducive to malignancy.

Histological, Immunohistochemical, and Biochemical Study of Experimentally Induced Fatty Liver in Adult Male Albino Rat and the Possible Protective Role of Pomegranate.

Nonalcoholic fatty liver disease is a major health problem and is considered the most common worldwide liver disease. Pomegranate has many biological activities and could modify the risk of hypercholesterolemia. The objective of the current research was to study the histological changes of experimentally induced fatty liver and possible protection by pomegranate. For this purpose, 50 adult male albino rats were divided into four groups, control group, pomegranate treated group that were given pomegranate juice for six weeks, fatty liver induced group that were fed on high fat diet for six weeks and protective group that were fed on high fat diet and received pomegranate juice for six weeks. Histological changes were detected in the fatty liver induced group in the form of disturbed hepatic architecture, dilatation and congestion of central veins, blood sinusoids and portal veins. Most of hepatocytes showed variable degrees of cytoplasmic vacuolation, mitochondrial structural changes, dilatation of endoplasmic reticulum in addition to nuclear structural changes like condensed chromatin, irregular shrunken nuclei and vacuolated nuclei. All these changes were associated with inflammatory cellular infiltrations, deposition of collagen fibers around the central vein, blood sinusoids, portal areas and in between the hepatocytes in addition to significant increase in number of hepatic stellate cells that was proved by electron microscope and confirmed by immunohistochemical study. Moreover, these structural changes were much less pronounced in animals treated with pomegranate either with or before receiving high fat diet. These findings suggested that pomegranate has a protective effect against experimentally induced fatty liver.

Hepatocyte-specific expression of constitutively active Alk5 exacerbates thioacetamide-induced liver injury in mice.

While Transforming growth factor-βs (Tgf-βs) have been known to play an important role in liver fibrosis through an activation of Hepatic Stellate Cells (HSC), their fibrotic role on hepatocytes in liver damage has not been addressed thoroughly. To shed more light on the hepatocyte-specific role of Tgf-β signaling during liver fibrosis, we generated transgenic mice expressing constitutively active Tgf-β type I receptor Alk5 under the control of albumin promoter. Uninjured mice with increased Tgf-β/Alk5 signaling in hepatocytes (caAlk5/Alb-Cre mice) did not show characteristics related to hepatocyte death, fibrosis and inflammation. When subjected to thioacetamide (TAA) treatment, caAlk5/Alb-Cre mice exhibited more severe liver injury, when compared to control littermates. After TAA administration for 12 weeks, an increase in pathological changes was evident in caAlk5/Alb-Cre livers, with higher number of infiltrating cells in the portal and periportal area. Immunohistochemistry for F4/80, myeloperoxidase and CD3 showed that there was an increased accumulation of macrophages, neutrophils and T-lymphocytes, respectively, in caAlk5/Alb-Cre livers. Coincidently, we observed an exacerbated liver damage as seen by increases in serum aminotransferase level and number of apoptotic hepatocytes in caAlk5/Alb-Cre mice. Sirius staining of collagen demonstrated that the fibrotic response was worsened in caAlk5/Alb-Cre mice. The enhanced fibrosis in mutant livers was associated with marked production of α-SMA-positive myofibroblast. Hepatic expression of genes indicative of HSC activation was greater in caAlk5/Alb-Cre mice. In conclusion, our data indicated that elevation of Tgf-β signaling via Alk5 in hepatocytes is not sufficient to induce liver pathology but plays an important role in amplifying TAA-induced liver damage.

Pathogenesis of Hepatic Deposited IgG-induced Liver Inflammation in SLE

Abstract Hepatic disorders are frequent in patients with systemic lupus erythematosus ( SLE), yet the etiology and pathogenesis of liver injury in SLE remain unclear. We used lupus-prone mice and established an animal model of intrahepatic deposited lupus IgG to investigate the pathogenesis of liver damage in SLE. We found that liver damage spontaneously developed in lupus-prone mice and that there were a large number of inflammatory cell infiltrates around the portal areas of the liver, apoptosis of hepatocytes, and IgG deposition in the liver. Liver inflammation developed in mice with intrahepatic injection of immune complexes (ICs) from SLE patients. We found that ICs stimulated Kupffer cells (KCs) to secrete TNF-α and mediated hepatic apoptosis. Moreover, IL-12 secreted by KCs induced the accumulation and activation of natural killer cells (NKs) in the liver. Further, IFN-γ produced by activated NKs directly mediated liver damage and also enhanced the TNF-α-mediated apoptotic pathway. The depletion of KCs and NKs abolished apoptosis induced by ICs in vivo, suggesting that KCs and NKs have a synergic effect on liver injury. Lupus IgG induced Syk activation, and inhibitor of sky suppressed skin inflammation triggered by lupus IgG. Taken together, our findings confirmed that the hepatic disease was mediated by SLE, which may provide insight into the cellular and molecular mechanisms of lupus hepatitis; further, our results may aid in future investigations of potential therapeutic strategies for the treatment and control of lupus-mediated injury in disease settings.

Biochemical and histopathological changes in sheep fed different detoxified karanj (Pongamia glabra) seed cake as partial protein supplements.

The study investigated the long-term effect of feeding processed solvent extracted karanj (Pongamia glabra) cake (SKC) on gross pathology and histopathological changes in some vital organs, and on the activities of serum enzymes in Jalauni lambs. Twenty-four male lambs were divided into 4 groups and allotted randomly to a soybean meal (SBM) based control (CON) and 3 treatment groups receiving concentrate mixtures, containing water washed (WW), 2.5% lime (LM) and 0.4% binder (BN) treated SKC replacing 50% nitrogen of SBM to meet the protein requirements. Blood was collected after 150 days from all the lambs and serum activities of aspartate aminotransferase (AST), alanine aminotransferase (ALT) and lactate dehydrogenase (LDH) were assayed. All lambs were sacrificed after 196 days, and vital organs screened for gross pathological lesions. The representative tissues of liver, intestine, parathyroid gland, testis, and epididymis were sampled, preserved in 10% formalin and processed to examine histopathological changes by staining with haematoxylin and eosin (HE). The serum enzyme activities of AST and ALT were similar in the treatments, but the activity of LDH was higher (P < 0.01) in processed SKC-fed groups than the control. The weight and size of the liver were decreased in BN group, while those of testes were decreased in both LM and BN groups. The histological sections of the testes and epididymis of lambs from LM group showed hypoplastic seminiferous tubules and empty ductules, respectively. The histological sections of the parathyroid gland in the same diet revealed smaller and non-secretory chief cells. The small intestine of lambs from BN group showed infiltration of mononuclear cells (MNC) in lamina propria with mild fibroplasia in intervillous areas. The histological section of liver from this group showed MNC infiltration in portal areas. The inclusion of water washed SKC in the concentrate mixture of lambs did not show gross pathological and histological alterations in the tissues in the vital organs; however, the activity of LDH was significantly (P = 0.001) elevated in processed SKC-fed groups than the control. Thus, feeding of water washed SKC in the concentrate at 225 g/kg for a longer period do not cause any adverse effect in lambs. This is supported by normal activities of serum enzymes and intact histological features in the tissues of liver, intestine, parathyroid gland and testis.

D2-40 Immunoreactivity of Lymphatic Vessels Endothelium and Representation of Lymphatic Vessels in the Liver of Human Fetuses of Different Gestational Age

Summary In the reference literature, there are a few studies on the development of the lymphatic system in the liver, especially human. This study aims to establish the presence, time of appearance, distribution and representation of expression D2-40 molecule – a marker of lymph vessels endothelial cells during the fetal period of the human liver development. The livers obtained from 20 human fetuses (10 male and 12 female), aged 12-37 gestational weeks, constituted our study material. Paraffin sections, 4 µm thick, were stained with hematoxylin and eosin for histological analysis, and with LSAB2/HRP method for immunohistochemistry using the D2-40 monoclonal antibody to mark lymphatic endothelial cells. The presence of lymphatic vessels was determined by morphometry, calculating their numerical and volume density. The study showed that expression of D2-40 molecule was absent in the liver lymphatic vessels in the first trimester of development, while in the second trimester intensive D2-40 immunoreactivity was observed in the lymph vessels of the liver capsule, and low D2-40 immunopositivity of the lymph vessels in large portal spaces. In the third trimester, intensive D2-40 immunoreactivity was observed in the lymph vessels of the liver capsule and in the endothelium of numerous lymphatic vessels of various shape and size, located in the smaller and larger portal areas. Volume and numerical density of lymphatic vessels in the portal areas of the liver during fetal development increased from the second to the third trimester of pregnancy, which was proportional to the increase in volume density of the hepatic portal spaces. Based on the obtained results, a conclusion may be drawn that the lymph vessels in the liver can be identified in the first half of the second trimester, and their number was growing proportionally by the end of pregnancy.