Cytoplasm Of Hepatocytes Research Articles

The low protein diet affects the nonspecific inflammatory response of middle-aged and old mice through mTOR.

To explore the effect of low protein diet on nonspecific inflammatory changes in mice during aging and related mechanisms. Thirty-two 14-month-old female KM mice were randomly divided into 4 groups: control group, low protein group, high protein group, high protein + rapamycin group. Hematoxylin-eosin (HE) staining was performed to observe the pathological changes of the liver. Immunohistochemistry of liver sections was performed to detect the expression of CD68 protein. HE staining of colon sections was performed to observe intestinal lymphocyte infiltration. The percentage of spleen CD4+ T and CD8+ T cells was detected by flow cytometry. The mTOR expression in the liver was detected by Western blot and immunohistochemistry. Compared with the control group, HE staining of liver tissue sections in high-protein group showed the cytoplasm of hepatocytes was loose and disordered, and the hepatic sinus was significantly expanded. Immunohistochemistry of the liver showed a significant increase in CD68 protein expression. Colorectal HE staining showed extensive lymphocyte infiltration. The number of CD4+ T and CD8+ T cells in spleen flow cytometry was significantly decreased (*p < 0.05). Western blot and immunohistochemistry detected a significant increase in mTOR expression in the liver (*p < 0.05, *p < 0.05). In the High Protein+Rapamycin group and Low-protein group, the time-dependent changes were reduced, the numbers of CD4+ T cell and CD8+ T cell in the spleen were significantly increased (*p < 0.05) and the expression of mTOR was significantly reduced (*p < 0.05). Low-protein diet is beneficial for delaying the non-specific inflammatory changes of liver and intestines in middle-aged and aged mice, and this effect may be achieved through down-regulation of mTOR.

Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues.

Silver nanoparticles (AgNPs) have been extensively used in commercial products, including textiles, cosmetics, and health care items, due to their strong antimicrobial effects. They also may be released into the environment and accumulate in the ocean. Therefore, AgNPs are the major source of Ag contamination, and public awareness of the environmental toxicity of Ag is increasing. Previous studies have demonstrated the bioaccumulation (in producers) and magnification (in consumers/predators) of Ag. Cetaceans, as the apex predators of ocean, may have been negatively affected by the Ag/Ag compounds. Although the concentrations of Ag/Ag compounds in cetacean tissues can be measured by inductively coupled plasma mass spectroscopy (ICP-MS), the use of ICP-MS is limited by its high capital cost and the requirement for tissue storage/preparation. Therefore, an autometallography (AMG) method with an image quantitative analysis by using formalin-fixed, paraffin-embedded (FFPE) tissue may be an adjuvant method to localize Ag distribution at the suborgan level and estimate the Ag concentration in cetacean tissues. The AMG positive signals are mainly brown to black granules of various sizes in the cytoplasm of proximal renal tubular epithelium, hepatocytes, and Kupffer cells. Occasionally, some amorphous golden yellow to brown AMG positive signals are noted in the lumen and basement membrane of some proximal renal tubules. The assay for estimating the Ag concentration is named the Cetacean Histological Ag Assay (CHAA), which is a regression model established by the data from image quantitative analysis of the AMG method and ICP-MS. The use of AMG with CHAA to localize and semi-quantify heavy metals provides a convenient methodology for spatio-temporal and cross-species studies.

Age features of the morpho-functional state of the liver in the distant terms of experimental diabetes mellitus.

Background. Between the most frequent complications of diabetes mellitus are diabetic hepatopathies, which are accompanied by a violation of the functional activity of the liver. Objective. Therefore, the purpose of our study was to establish the morpho-functional changes of the liver of 24 months old rats in the distant terms of streptozotocine diabetes mellitus. Methods. The study was conducted on 24 male Wistar rats that had reached the age of 24 months. The animals were divided into 2 groups: experimental (15 animals) and control (9 animals). For streptozotocine diabetes mellitus modelling, animals of the experimental group were administered intraperitoneally with streptozotocin in a dose of 5 mg per 100 g of body weight. Biochemical, histological (coloring of hematoxylin-eosin, Shabadash, Sudan III) and electron-microscopic methods of research were used. The material was taken on the 42, 56 and 70 days of the streptozotocine diabetes mellitus course. Results. At the 42 day of streptozotocine diabetes mellitus we detect the unevenness of the coloration of the hepatocyte cytoplasm due to the presence of small vacuoles, indicating the development of vacuolic dystrophy. At the 56 day of streptozotocine diabetes mellitus, both on the optical and electron microscopic levels, we find the signs of lipid, vacuolic and balloon dystrophy. In hepatocytes there are numerous lipid drops, which may indicate the development of fatty dystrophy. At the 70 day of streptozotocine diabetes mellitus, a significant polymorphism of morphological changes in hepatocytes is observed. Some of them show the signs of vacuolation, others - balloon dystrophy. We observe that separate hepatocytes are in the state of colic necrosis, others - are in a state of apoptosis. Starting from the 42 day of streptozotocine diabetes mellitus, we observe an increased level of activity of enzymes of the cytolysis of hepatocytes, both in blood serum and in liver homogenates, in comparison with the control group of animals, and the reduction of the Ritis coefficient. Conclusion. Thus, in the distant terms of streptozotocine diabetes mellitus (42-70 days), we observe the morpho-functional changes in the liver, which are manifested: vacuolar and hydropic dystrophies, colic-type necrosis, apoptosis. Such structural changes in the liver lead to a violation of its function, which is confirmed by a significant increase in the level of alanine aminotransferase and aspartate aminotransferase and simultaneous decrease in the Ritis coefficient.

28-Day Oral Chronic Toxicity Study of Arctigenin in Rats.

Arctium lappa (burdock) is the most popular daily edible vegetable in China and Japan because of its general health tonic effects. Previous studies focused on the beneficial role of Arctigenin but neglected its potential side-effects and toxicities. In the present study, the sub-chronic toxicity profile of Arctigenin following 28 days of consecutive exposure was investigated in rats. The results showed that during the drug exposure period, Arctigenin-12 mg/kg administration resulted in focal necrosis and lymphocytes infiltration of heart ventricular septal muscle cells. In the kidney cortical zone, the renal tubular epithelial cells were swollen, mineralized, and lymphocyte infiltrated. In the liver, the partial hepatocyte cytoplasm showed vacuolation and fatty changes, focal necrosis, and interstitial lymphocyte infiltration. In the rats that underwent 36 mg/kg/day administration, there was bilateral testis and epididymis atrophy. In the lung and primary bronchus, erythrocytes and edema fluid were observed. Changes of proestrus or estrus were observed in the uterus, cervix, and vagina intimal epithelial cells. Lymphocytic focal infiltration occurred in the prostate mesenchyme. The high dosage of Arctigenin only decreased the body weight at day 4. At the end of the recovery period, histopathological changes were irreversible, even after withdrawal of the drug for 28 days. Focal necrosis still existed in the heart ventricular septal muscle cells and hepatocytes. Lymphocyte infiltrations were observed in the heart, renal cortex, hepatocyte, and pancreas exocrine gland. Meanwhile, atrophy occurred in the testicles and pancreas. In addition, in the Arctigenin-12 mg/kg group, creatinine (CREA) and brain weight were both significantly increased. The toxicokinetical study demonstrated that Arctigenin accumulated in the organs of rats. The food consumption, hematological, and biochemical parameters were not associated with the above results. These contradictory results might result from the lesions induced by Arctigenin, which were not sufficiently serious to change the parameters. These results suggest that Arctium lappa should be consumed daily with caution because of the potential toxicity induced by Arctigenin. According to all results, the lowest observed adverse effect level (LOAEL) was induced by 12 mg/kg daily exposure to Arctigenin, and the No-observed-adverse-effect-level (NOAEL) should be lower than 12 mg/kg.

БИОХИМИЧЕСКИЕ И МОРФОМЕТРИЧЕСКИЕ ПОКАЗАТЕЛИ НЕКОТОРЫХ ОРГАНОВ И ТКАНЕЙ ГИБРИДА ТИЛЯПИИ (OREOCHROMIS SPP.) ПРИ ВЫРАЩИВАНИИ С ИСПОЛЬЗОВАНИЕМ ПРЕПАРАТА ЭС-2

The article studies the effect of addition into the feed of Sapropel extract (ES-2 preparation) on the intensity of lipid peroxidation in the liver and gills of hybrid tilapia ( Oreochromis spp. ), as well as on the morphofunctional state of its liver. Sapropel extract caused a decrease in the content of TBA-reactants in the tissues of tilapia liver by 17% compared to the control group. In gills the bioadditive resulted in the increased content of peroxide products by 24%. The introduction of ES-2 in fish feed resulted in reduction of spontaneous and ascorbate-dependent lipid peroxidation rate in the liver by 18%. In the gills of fish, under the influence of Sapropel, the rate of spontaneous lipid peroxidation increased by 27%, the rate of ascorbate-dependent lipid peroxidation - by 23%. The change in the intensity of peroxide processes under the influence of the fodder additive in fish organs is tissue-specific: antioxidant effect was recorded in the liver, prooxidant effect was observed in the gills. The introduction of the Sapropel extract does not lead to a change in the volume of liver nuclei in the test groups of tilapia, while the average cell volume in the experimental group was 37% lower than in the control group. The decrease in cell volume led to the increase in the nuclear-cytoplasmic ratio by 1.9 times in the experimental group compared to the control group. Hepatocyte cytoplasm volume decrease and nuclear-cytoplasmic ratio increase due to addition of ES-2 preparation into productive feed of hybrid tilapia would indicate a rise of functional activity of liver cells.

Different roles of FAT10, FOXO1, and ADRA2A in hepatocellular carcinoma tumorigenesis in patients with alcoholic steatohepatitis (ASH) vs non-alcoholic steatohepatitis (NASH)

Hepatocellular carcinoma (HCC) is the fifth most common cancer and the second leading cause of cancer related deaths worldwide. Among others, non-alcoholic steatohepatitis (NASH) and alcoholic steatohepatitis (ASH) are the two major risk factors as both of them may develop cirrhosis and hepatocellular carcinoma (HCC) if left untreated. However, patients with NASH progress to HCC at a rate around 0.5% annually, while 3–10% ASH patients may progress to HCC annually. The present study is to demonstrate the molecular differences in oncogenesis pathway between NASH and ASH. By using immunofluorescence study and quantitating the fluorescence intensity morphometrically in liver biopsied specimens from NASH and ASH patients, the protein expression of candidate molecules within hepatocytes cytoplasm are studied, including two HCC-related molecules FAT10 and FOXO1, and one GPCR pathway related molecule ADRA2A. Compared with the control group patients, the expression levels of all the molecules were upregulated in the ASH group of patients (p < 0.001 in all molecules), while FAT10 and ADRA2A were upregulated, FOXO1 did not change in the NASH group of patients. The most important finding is that compared with the ASH group of patients, the expression levels of all three molecules were significantly lower than in the NASH group of patients (p < 0.001 in all molecules). These results confirmed our previous finding that there are significant differences of molecules change in ASH compared to NASH. Thus, we conclude that there are significantly different molecules and pathways involved during the pathogenesis of HCC development in ASH compared to NASH which could help explain why the tumorigenic rate is different in ASH and NASH.

Autophagy in Hepatocytes during Distant Tumor Growth.

Structural changes in the liver of CBA mice were studied during the development of experimental hepatocarcinoma-29 inoculated into the hip. A decrease in the volume density of hepatocyte cytoplasm, mitochondria, endoplasmic reticulum, and lipid inclusions and an increase in the volume density of lysosomal structures during tumor growth were observed. All stages of intracellular autophagy were recorded by the method of electron microscopy. These stages included the appearance of autophagosomes, autophagolysosomes, and secondary lysosomes in the hepatocyte cytoplasm. Fragments of cytoplasm, glycogen rosettes, mitochondria, and fragments of endoplasmic reticulum with ribosomes were found in autophagosomes. The obtained data indicate the development of non-selective autophagy in the liver during distant tumor growth in aimed at the maintenance of intracellular homeostasis in hepatocytes and energy and trophic homeostasis of organism.

Diagnosis of Q Fever and Brucellosis in Aborted Ovine Fetuses by Microbiological, Pathological and Immunohistochemical Methods

Abstract Brucellosis and Q fever, two zoonoses, are important causes of abortion in ruminants, as well as economically significant diseases caused by a gram-negative bacterium. Determination of these diseases is therefore of great importance. In this study, the organs of 35 naturally infected and aborted ovine fetuses were examined for the presence of changes resulting from infections by Brucella melitensis and Coxiella burnetii, according to macroscopic, bacteriological, histopathological and immunohistochemical methods. B. melitensis was observed in 21 cases, and C. burnetii was observed in 8 cases of the aborted ovine fetuses, and these were determined with immunohistochemical methods. Brucellosis was observed in 18 of the aborted ovine fetuses, and this was determined by microbiological methods. Negative (-) results were found for all of the other fetuses. The Brucella antigen was determined to be localized as intracytoplasmic in mainly alveolar macrophages, bronchi, bronchioles, glandular epithelial cells around bronchial glands, neutrophils, hepatocytes and Kupffer cells. The Coxiella antigen was found to be localized in the alveolar macrophages in the lungs, bronchi, bronchioles and alveolus, and in the cytoplasms of bronchial gland epithelial cells, and in the cytoplasms of hepatocytes and Kupffer cells in the liver. Immunohistochemical and microbiological diagnoses of brucellosis and coxiellosis were compared; it was concluded that immunohistochemical methods were more safely applied than microbiological methods.

Innate Immunity to Enteric Hepatitis Viruses.

Although hepatitis A virus (HAV) and hepatitis E virus (HEV) are both positive-strand RNA viruses that replicate in the cytoplasm of hepatocytes, there are important differences in the ways they induce and counteract host innate immune responses. HAV is remarkably stealthy because of its ability to evade and disrupt innate signaling pathways that lead to interferon production. In contrast, HEV does not block interferon production. Instead, it persists in the presence of an interferon response. These differences may provide insight into HEV persistence in immunocompromised patients, an emerging health problem in developed countries.

Relationship among biomarkers of iron metabolism and severity of underlying nonalcoholic steatohepatitis (NASH)

Background: In obese subjects with nonalcoholic steatohepatitis (NASH), hepatic iron load may be associated with the risk of toxicity, acting as a comorbidity factor involved in fibrogenesis and carcinogenesis. To evaluate the relationship among biomarkers of iron metabolism and the severity of NASH in adult obese subjects. Methods: Eighty-eight adult subjects with histological diagnosis of non-alcoholic fatty liver disease (NAFLD) were evaluated in a prospective study for the biomarkers of iron metabolism: serum iron, transferrin and ferritin, as well as transferrin saturation index (TSI). Iron deposition in the liver, the intensity of steatosis and fibrosis were assessed in liver biopsies. Results: Increased serum levels of ferritin were found in 31/88 (35.2%) subjects and 12/88 (13.6%) had values of TSI above 45%. Histological evaluation of liver biopsies showed the presence of fibrosis in 48/88 cases (54.5%) being 23.8% mild and 30.7% moderate/severe. Analysis of stainable iron in liver biopsies by Perls method, demonstrated fine granular deposits in the cytoplasm of hepatocytes in 17/88 biopsies (19.3%). There was no association between the presence of iron deposits in the liver parenchyma and the intensity of fibrosis. Multiple logistic regression analysis identified type 2 diabetes mellitus (T2DM) or fasting glucose >100 mg/dL and dyslipidemia as factors independently associated with fibrosis. Conclusions: The findings from our study did not confirm the association between the presence of iron in the hepatic parenchyma and the severity of NASH in obese subjects.

Apple Cider Vinegar (as a Prophetic Medicine Remedy) Exerts Tissue-protective Effects in Streptozotocin-induced Diabetes in Animals

Background: Diabetes mellitus (DM) was reported to be related to occurrence of both biochemical and histopathological changes in hepatic tissue. DM-induced tissue damage and decreased immunity may favor carcinogenesis and tumor development in pediatric practice and adults. In this study, we aimed at investigating the hepatoprotective and tissue-protective effects of apple cider (ACV) against DM-induced hepatic derangements over a relatively long period (30 days). Effects were evaluated using streptozotocin (STZ)-induced DM in rats as an experimental model. Materials and methods: DM was experimentally caused by injecting one dose of STZ (65 mg/kg given through intraperitoneal injection. Thirty wistar rats were categorized into three experimental groups: control group, STZ-treated group and STZ plus ACV treated group 2 ml/kg BW). Animals were sacrificed 30 days post treatment. Results: Our study revealed that ACV administration induced hepatoprotective effects in diabetic rats. Biochemically, ACV reduced significantly serum glycemic values (glucose, total cholesterol, low density lipoprotein-cholesterol while high density lipoprotein cholesterol significantly increased). STZ-induced DM in rats was associated with marked fatty changes hepatocytes cytoplasm evidenced by accumulation of lipid droplets and inflammatory changes e.g. lymphocytic infiltration. That was confirmed by Electron microscopic evaluation that showed aggregations of polymorphic mitochondria, loss of mitochondrial cristae and condensed mitochondrial matrices. Hepatocytes cytoplasm showed vacuolations with a large number of variable-sized lipid droplets. Hepatocytes rough endoplasmic reticulum got distorted and fragmented into smaller stacks. ACV proved effective in reversing all these metabolic changes. ACV-treated hepatocytes exhibited minimal STZ-induced toxic effects. ACV enhanced recovery of the injured hepatocytes and normalized serum biochemical values. In prophetic medicine, Prophet Muhammad (Allah's apostle peace be upon him) strongly advised eating and considered it as the best edible as narrated in the prophetic hadith: vinegar is the best edible. Conclusion: ACV is promising as a food additive or as a part of diet in diabetic patients particularly in early stages of DM. ACV slows down DM progress and causes hepatoprotection against the metabolic damages resulting from STZ- induced diabetes mellitus

Zebrafish abcb11b mutant reveals strategies to restore bile excretion impaired by bile salt export pump deficiency.

Collectively, these data suggest a model that rapamycin rescues BSEP-deficient phenotypes by prompting alternative transporters to excrete bile salts; multidrug resistance protein 1 is a candidate for such an alternative transporter. (Hepatology 2018;67:1531-1545).

Microarray analysis reveals Tmub1 as a cell cycle-associated protein in rat hepatocytes.

Transmembrane and ubiquitin-like domain containing protein 1 (Tmub1), formerly known as hepatocyte odd protein shuttling (HOPS) has been recognized as a ubiquitously expressed shuttling protein that moves between the nucleus and cytoplasm in hepatocytes. Tmub1 is involved in liver regeneration and functions as a bridging protein in tumor cell proliferation. To investigate the transcriptional profile and potential biological processes affected by Tmub1 expression in normal rat hepatocytes, microarray and bioinformatics experiments were used to identify 127 mRNAs differentially expressed between Tmub1-overexpression, Tmub1-knockdown and normal BRL-3A cells (fold-change ≥2.5). The expression levels of 17 key node genes associated with the cell cycle were confirmed by reverse transcription-quantitative polymerase chain reaction analysis. Flow cytometry, 5-Ethynyl-20-deoxyuridine, Cell Counting Kit-8 and western blotting experiments revealed the effects on the cell cycle and the inhibition of proliferation in BRL-3A cells overexpressing Tmub1. Further co-immunoprecipitation assays demonstrated that Tmub1 interacts with cyclin A2 during the cell cycle and that the overexpression of Tmub1 may postpone cyclin A2 and cyclin B1 degradation in the M phase. The results of the present study indicated that Tmub1 functions as a cell proliferation inhibitor and cell cycle-associated protein.

Arkadia protein expression is reduced in the liver during the progression of hepatic fibrosis.

Arkadia is able to degrade key signaling molecules in the transforming growth factor (TGF)-β1 signaling pathway; however, the expression of Arkadia in the liver during development and progression of TGF-β1/Smad signaling-regulated hepatic fibrosis remains to be elucidated. The present study aimed to examine Arkadia expression in the livers of two rat models of hepatic fibrosis induced by bile duct ligation and carbon tetrachloride intoxication, and in human liver samples from patients with hepatic fibrosis. Expression was analyzed by quantitative polymerase chain reaction, immunohistochemistry and western blot analysis. The results indicated that Arkadia was predominantly expressed in the cytoplasm of cholangiocytes and hepatocytes. The protein expression levels of Arkadia were significantly decreased in fibrotic livers, whereas the mRNA expression levels of Arkadia were significantly increased in fibrotic livers compared with in nonfibrotic livers. In conclusion, these data indicated that Arkadia may regulate the pathogenesis and progression of hepatic fibrosis.

Potential effect of Nigella sativa against Diethylnitrosamine-induced hepatocarcinogenesis in rats

Objective of the investigation was the study of potential protective effects of the watery extract of Nigella sativa against diethylnitrosamine induced hepatocarcinogenesis in rats. N. sativa was administered to rats for protection against diethylnitrosamine-induced hepatocarcinogenesis. It was administered prior to, simultaneously with or after injection of diethylnitrosamine. Five groups of Wister rats were used. Group A was administered diethylnitrosamine and N. sativa simultaneously, group B was administered only diethylnitrosamine and group C received only N. sativa. These three groups were maintained for up to eight weeks. Group D received N. sativa six weeks after administration ofdiethylnitrosamine,while group E (“protective group”) received N. sativa on day 1 and diethylnitrosamine six weeks later. These two groups were maintained for up to 12 weeks. All rats were subjected to partial hepatectomy to enhance carcinogenesis. P-isoform of glutathione s transferase (GST-P) was detected in the cytoplasm and nuclei of hepatocytes. The number of GST-P positive foci was significantly smaller in test groups (A, D, E), particularly in groups A and E, when compared with to those in group B, indicating that N. sativa has protective effects against diethylnitrosamine induced liver cancer in rats, even in the very early stages of hepatocarcinogenesis.

The Role of Necroptosis, Apoptosis, and Inflammation in Fowl Cholera-Associated Liver Injury in a Chicken Model.

Fowl cholera resulting from infection with Pasteurella multocida causes huge economic losses in the poultry industry. Necrotic hepatitis is reported to be a significant lesion associated with fowl cholera in chickens. Clarifying the underlying molecular mechanism of hepatic injury caused by P. multocida infection is needed to develop new strategies to control fowl cholera. Pasteurella multocida Q (the standard reference strain) and P. multocida 1G1 (a clinical strain) were used to infect healthy laying hens. Clinical signs were observed and gross lesions in livers were observed postmortem. Histologic lesions and the localization and expression of protein molecules associated with necroptosis, apoptosis, and inflammation in hepatic tissues were examined by hematoxylin and eosin staining and immunohistochemistry. Western blot analysis was used to determine the expression of liver injury-related genes. Necroptotic molecules such as RIPK1 (receptor interaction protein kinases 1), RIPK3 (receptor interaction protein kinases 3), and MLKL (mixed lineage kinase domain-like protein) were observed by immunostaining primarily in the cytoplasm of hepatocytes within or around necrotic foci, and inflammatory mediators HMGB1 (high-mobility group box 1) and IL-6 (interleukin-6) were found in the cytoplasm of heterophils, monocytes/macrophages, and hepatic sinusoids. In addition, MMP9 (matrix metalloproteinase 9) and TIMP1 (tissue inhibitor of metalloproteinase 1) were observed in hepatic parenchymal cells, inflammatory cells, and interstitial spaces, whereas the apoptotic effector molecule caspase-3 (cysteine-containing aspartic proteolytic enzymes 3) was mainly found in hepatocytes. The expression of RIPK1, RIPK3, and MLKL was significantly higher in the infected chickens than in the controls. HMGB1 and IL-6 protein levels were also increased in infected chickens relative to those in controls. Both MMP9 and TIMP1 were highly expressed in infected chickens. In addition, caspase-3 protein levels were significantly elevated in infected chickens. Necroptosis, apoptosis, and inflammation played a significant role in hepatic injury caused by P. multocida.

Possible role of Arthrospira platensis in reversing oxidative stress-mediated liver damage in rats exposed to lead

Environmental pollutants, particularly metallic elements, mobilized and released into the environment, eventually accumulate in the food chain and thus pose a serious threat to human and animal health. In the present study, the role of Arthrospira (Spirulina platensis; SP) as a protector against oxidative stress-mediated liver damage induced by an exposure to lead acetate (LA; as a metallic pollutant) was assessed. To achieve this aim, rats were orally administered with 300 mg/kg bw SP for 15 days, before and concurrently with an intraperitoneal injection of 50 mg/kg bw LA (6 injections throughout 15 days). As a result, co-administration of SP with LA reduced the amount of lead that accumulated in both blood and liver tissue of the exposed rats and minimized the increased levels of lipid peroxidation, protein oxidation, DNA oxidative damage, and liver enzyme endpoints. In addition, because of SP administration, the levels of depleted biomarkers of antioxidant status and total antioxidant capacity in LA-exposed rats improved. Moreover, SP protected the liver tissue against the changes caused by LA exposure and also decreased the reactivity of HSP70 in the cytoplasm of hepatocytes. Collectively, our data suggest that SP has a potential use as a food supplement in the regions highly polluted with heavy metals such as lead.

CRISPR/Cas9 Engineering of Adult Mouse Liver Demonstrates That the Dnajb1–Prkaca Gene Fusion Is Sufficient to Induce Tumors Resembling Fibrolamellar Hepatocellular Carcinoma

Fibrolamellar hepatocellular carcinoma (FL-HCC) is a primary liver cancer that predominantly affects children and young adults with no underlying liver disease. A somatic, 400 Kb deletion on chromosome 19 that fuses part of the DnaJ heat shock protein family (Hsp40) member B1 gene (DNAJB1) to the protein kinase cAMP-activated catalytic subunit alpha gene (PRKACA) has been repeatedly identified in patients with FL-HCC. However, the DNAJB1-PRKACA gene fusion has not been shown to induce liver tumorigenesis. Weused the CRISPR/Cas9 technique to delete in mice the syntenic region on chromosome 8 to create a Dnajb1-Prkaca fusion and monitored the mice for liver tumor development. We delivered CRISPR/Cas9 vectors designed to juxtapose exon 1 of Dnajb1 with exon 2 of Prkaca to create theDnajb1-Prkaca gene fusion associated with FL-HCC, or control Cas9 vector, via hydrodynamic tail vein injection to livers of 8-week-old female FVB/N mice. These mice did not have any other engineered genetic alterations and were notexposed to liver toxins or carcinogens. Liver tissues werecollected 14months after delivery; genomic DNA was analyzed by PCR to detect the Dnajb1-Prkaca fusion, and tissues were characterized by histology, immunohistochemistry, RNA sequencing, and whole-exome sequencing. Livers from 12 of the 15 mice given the vectors to induce the Dnajb1-Prkaca gene fusion, but none of the 11 mice given the control vector, developed neoplasms. The tumors contained the Dnajb1-Prkaca gene fusion and had histologic and cytologic features of human FL-HCCs: large polygonal cellswith granular, eosinophilic, and mitochondria-rich cytoplasm, prominent nucleoli, and markers of hepatocytes and cholangiocytes. In comparing expression levels of genes between the mouse tumor and non-tumor liver cells, we identified changes similar to those detected in human FL-HCC, which included genes that affect cell cycle and mitosis regulation. Genomic analysis of mouse neoplasms induced by the Dnajb1-Prkaca fusion revealed a lack of mutations in genes commonly associated with liver cancers, as observed in human FL-HCC. Using CRISPR/Cas9 technology, we found generation of the Dnajb1-Prkaca fusion gene in wild-type mice to be sufficient to initiate formation of tumors that have many features of human FL-HCC. Strategies to block DNAJB1-PRKACA might be developed as therapeutics for this form of liver cancer.

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.

Endotoxemia-mediated activation of acetyltransferase P300 impairs insulin signaling in obesity

Diabetes and obesity are characterized by insulin resistance and chronic low-grade inflammation. An elevated plasma concentration of lipopolysaccharide (LPS) caused by increased intestinal permeability during diet-induced obesity promotes insulin resistance in mice. Here, we show that LPS induces endoplasmic reticulum (ER) stress and protein levels of P300, an acetyltransferase involved in glucose production. In high-fat diet fed and genetically obese ob/ob mice, P300 translocates from the nucleus into the cytoplasm of hepatocytes. We also demonstrate that LPS activates the transcription factor XBP1 via the ER stress sensor IRE1, resulting in the induction of P300 which, in turn, acetylates IRS1/2, inhibits its association with the insulin receptor, and disrupts insulin signaling. Pharmacological inhibition of P300 acetyltransferase activity by a specific inhibitor improves insulin sensitivity and decreases hyperglycemia in obese mice. We suggest that P300 acetyltransferase activity may be a promising therapeutic target for the treatment of obese patients.