glycogen and Chemical-and-Drug-Induced-Liver-Injury--Chronic

Hepatocytes from human bone marrow-derived mesenchymal stem cells (hBM-MSCs) are expected to be a useful source for cell transplantation. However, relatively low efficiency and repeatability of hepatic differentiation of human BM-MSCs remains an obstacle for clinical translation. Hepatocyte nuclear factor 4 alpha (HNF4α), a critical transcription factor, plays an essential role in the entire process of liver development. In this study, immortalized hBM-MSCs, UE7T-13 cells were transduced with a lentiviral vector containing HNF4α. The typical fibroblast-like morphology of the MSCs changed, and polygonal, epithelioid cells grew out after HNF4α transduction. In hepatocyte culture medium, HNF4α-transduced MSCs (E7-hHNF4α cells) strongly expressed the albumin (ALB), CYP2B6, alpha-1 antitrypsin (AAT), and FOXA2 mRNA and exhibited morphology markedly similar to that of mature hepatocytes. The E7-hHNF4α cells showed hepatic functions such as Indocyanine green (ICG) uptake and release, glycogen storage, urea production and ALB secretion. Approximately 28% of E7-hHNF4α cells expressed both ALB and AAT. Furthermore, these E7-hHNF4α cells via superior mesenteric vein (SMV) injection expressed human ALB in mouse chronic injured liver. In conclusion, this study represents a novel strategy by directly inducing hepatocyte-like cells from MSCs.

Topics: Albumins; alpha 1-Antitrypsin; Animals; Antibodies, Monoclonal; Antibodies, Monoclonal, Murine-Derived; Bone Marrow Cells; Cell Differentiation; Cell Line, Transformed; Chemical and Drug Induced Liver Injury, Chronic; Cytochrome P-450 CYP2B6; Gene Expression Regulation; Genetic Vectors; Glycogen; Hepatocyte Nuclear Factor 3-beta; Hepatocyte Nuclear Factor 4; Hepatocytes; Humans; Lentivirus; Male; Mesenchymal Stem Cells; Mice; RNA, Messenger; Signal Transduction; Transduction, Genetic; Urea

Allopurinol is an inhibitor of xanthine oxidase. The aim of this work was to evaluate the efficacy of allopurinol to reverse the experimental cirrhosis induced by CCl4. Rats received CCl4 for 8 weeks, and immediately after allopurinol was administered for 4 weeks more. Allopurinol reversed all markers of liver damage and oxidative stress to normal values, restoring the metabolic capacity of the liver. Chronic injury by CCl4 induced significant overexpression of profibrogenic cytokine TGF-β, while allopurinol decreased this production and consequently decreased the collagen content. Moreover, allopurinol is capable of partially inhibiting NF-κB. These findings suggest that allopurinol is capable of reversing the cirrhosis induced by CCl4, modulating oxidative stress, TGF-β expression and NF-κB nuclear translocation.

Topics: Allopurinol; Animals; Carbon Tetrachloride; Chemical and Drug Induced Liver Injury, Chronic; Collagen; Glutathione; Glutathione Disulfide; Glycogen; Lipid Peroxidation; Liver; Male; NF-kappa B; Oxidative Stress; Rats; Rats, Wistar; Transforming Growth Factor beta; Xanthine Oxidase

The goal of the study was to examine the state of primary hepatocytes of rats with toxic hepatitis induced by combination of CCl4 and ethanol. Fluorescent immunocytochemical analysis demonstrated that normal and pathologic hepatocytes in culture formed actin cytoskeleton, cell-cell and cell-matrix contacts. To investigate morphology and localization of mitochondria the hepatocytes were stained with Rhodamine 123. Glycogen and DNA contents in hepatocytes were determined by fluorescent cytophotometry during the lifetime of the culture. Cells were maintained for 5 days, and there were no changes in ploidy distribution observed. The mean ploidy was not changed too. Thus hepatocytes of different ploidy demonstrated similar survival rate. The glycogen content was 50% higher in experimental group compared to the control. The glycogen content decreased in control and cyrrotic hepatocytes after collagenase isolation. It has been found that the control hepatocytes accumulated glycogen within 3 days. On the contrary, the glycogen levels remained to be low in the pathologic hepatocytes.

Topics: Animals; Carbon Tetrachloride; Cells, Cultured; Chemical and Drug Induced Liver Injury, Chronic; Disease Models, Animal; Ethanol; Glycogen; Hepatocytes; Male; Mitochondria; Rats; Rats, Wistar

Topics: Animals; Bilirubin; Carbon Tetrachloride Poisoning; Chemical and Drug Induced Liver Injury, Chronic; Glycogen; Liver; Male; Malondialdehyde; Rats; Soybean Proteins