epidermal-growth-factor and Liver-Failure--Acute

Liver regenerates following surgical removal and after drug-induced liver injury (DILI). However, most of the mechanisms of liver regeneration were identified using partial hepatectomy (PHX) model rather than using DILI models. We compared mechanisms of liver regeneration following PHX and after acetaminophen (APAP) overdose, a DILI model, using transcriptomic approach. Kinetics of hepatocyte proliferation and global gene expression profiles were studied in male C57BL/6J mice either subjected to PHX or following APAP overdose. Liver regeneration was much more synchronized after PHX as compared to APAP overdose. Transcriptomics analysis revealed activation of common upstream regulators in both models including growth factors HGF, EGF and VEGF; and cytokines IL6 and TNFα. However, magnitude of activation and temporality was significantly differed between the two models. HGF and VEGF showed similar activation between PHX and APAP but activation of EGF was significantly stronger in the APAP model. Activation of IL6 and TNFα transcriptional programs was delayed but remarkably higher in APAP. These dissimilarities could be attributed to inherent differences in the two models including significant injury and inflammation exclusively in the APAP model. This study highlights need to study mechanisms of liver regeneration after DILI separately from the mechanisms of regeneration PHX.

Topics: Acetaminophen; Animals; Cell Proliferation; Chemical and Drug Induced Liver Injury; Epidermal Growth Factor; Gene Expression Profiling; Hepatectomy; Hepatocytes; Liver; Liver Failure, Acute; Liver Regeneration; Male; Mice; Mice, Inbred C57BL; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A

Engineered hepatic tissue (EHT) is considered as a promising strategy for healing acute liver failure (ALF), therefore, in the present study we evaluated the therapeutic potential of the EHT which engaged with bone marrow mesenchymal cells (BMSCs) derived hepatocytes (BMSCs—Hepas) in ALF rats. After characterization of isolated BMSCs, we seeded passage 3 BMSCs which have being cultured in medium containing 20 ng/ml hepatocyte growth factor (HGF) and 10 ng/ml epidermal growth factor (EGF) for 14 days on three scaffolds individually in Transwell system, and then cultured for more than 3 days to construct three kinds of EHT named EHT1, EHT2, and EHT3. Based on morphology and urea production assays, we chose an optimal one and transplanted it into ALF rat with 90% subtotal hepatectomy and assessed its therapeutic potential by survival time, hepatic encephalopathy score (HES) and related liver function test. The remnant liver was acquired, sectioned and identified by con-focal scanning microscopy. The isolated cells possessed basic properties of BMSCs, when cultured in hepatogenic medium for 2 weeks, BMSCs would restore to the functional properties of primary rats' hepatocytes, expressing albumin (ALB) and alpha fetoprotein (AFP) simultaneously. Transplantation of EHT3 significantly prolonged the survival time, increased HES, and ameliorated the liver function. BMSC will be a newly cell source for the construction of EHT. Importantly, the EHT transplantation may be an effective strategy to treat ALF in clinic.

Topics: Albumins; alpha-Fetoproteins; Animals; Bone Marrow Cells; Cell Transdifferentiation; Cell- and Tissue-Based Therapy; Cells, Cultured; Cellular Reprogramming; Epidermal Growth Factor; Hepatectomy; Hepatocyte Growth Factor; Hepatocytes; Liver; Liver Failure, Acute; Male; Mesenchymal Stem Cells; Rats; Rats, Sprague-Dawley; Tissue Engineering; Tissue Scaffolds

The aims of this study are to explore the effects of epidermal growth factor (EGF) on hepatocyte proliferation in presence of plasma from patients with acute liver failure (ALF).. HepG2 cells were cultured with 50% plasma from patients with ALF for 6, 12, 24, 48 and 72h with or without different concentrations of EGF. Cell proliferation was determined by the MTT assay and intracellular cyclin D1, cyclin-dependent kinase 4 (CDK4) expressions were analyzed by western blotting.. The proliferation of HepG2 cells was significantly inhibited by treatment with plasma from patients with ALF from 12 to 72 h. Intracellular expression of cyclin D1 and CDK4 was also markedly down-regulated. 5ng/ml, 10ng/ml and 20ng/ml EGF dose dependently induced HepG2 proliferation in presence of plasma from normal control, but only 20ng/ml EGF showed a transient promoting effect on proliferation of HepG2 cells in presence of plasma from patients with ALF.. Plasma from patients with ALF inhibits HepG2 cell proliferation via downregulation of cyclin D1 and CDK4 expression. Plasma from patient with ALF dampens HepG2 cells to EGF induced proliferation response.

Topics: Adolescent; Adult; Apoptosis; Carcinoma, Hepatocellular; Case-Control Studies; Cell Proliferation; Cyclin D1; Cyclin-Dependent Kinase 4; Dose-Response Relationship, Drug; Epidermal Growth Factor; Female; Hep G2 Cells; Humans; Liver Failure, Acute; Liver Neoplasms; Male; Middle Aged; Time Factors

To investigate whether the function of hepatocytes co-cultured with bone marrow mesenchymal stem cells (MSCs) could be maintained in serum from acute-on-chronic liver failure (ACLF) patients.. Hepatocyte supportive functions and cytotoxicity of sera from 18 patients with viral hepatitis B-induced ACLF and 18 healthy volunteers were evaluated for porcine hepatocytes co-cultured with MSCs and hepatocyte mono-layered culture, respectively. Chemokine profile was also examined for the normal serum and liver failure serum.. Hepatocyte growth factor (HGF) and Tumor necrosis factor; tumor necrosis factor (TNF)-α were remarkably elevated in response to ACLF while epidermal growth factor (EGF) and VEGF levels were significantly decreased. Liver failure serum samples induced a higher detachment rate, lower viability and decreased liver support functions in the homo-hepatocyte culture. Hepatocytes co-cultured with MSCs could tolerate the cytotoxicity of the serum from ACLF patients and had similar liver support functions compared with the hepatocytes cultured with healthy human serum in vitro. In addition, co-cultured hepatocytes maintained a proliferative capability despite of the insult from liver failure serum.. ACLF serum does not impair the cell morphology, viability, proliferation and overall metabolic capacities of hepatocyte co-cultured with MSCs in vitro.

Topics: Animals; Bone Marrow Cells; Cell Proliferation; Cell Survival; Coculture Techniques; End Stage Liver Disease; Epidermal Growth Factor; Female; Hepatocyte Growth Factor; Hepatocytes; Humans; Liver Failure, Acute; Male; Mesenchymal Stem Cells; Serum; Swine; Tumor Necrosis Factor-alpha; Vascular Endothelial Growth Factor A