Background Non-alcoholic steatohepatitis (NASH) is a chronic disease characterized by inflammation, steatosis, and liver fibrosis. The liver is particularly affected by alterations in lipid metabolism. Our aim was to evaluate the effect of β-hydroxyphosphocarnitine (β-HPC) on NASH induced in rats. Methods NASH was produced via the ad libitum daily chronic administration of a fructose solution (400 kcal) for 9 weeks, an oral dose of fat solution (16 kcal) for 7 weeks and a subcutaneous injection of CCl4 (30%) two times a week for 2 weeks to Wistar rats. To evaluate the effect of β-HPC, a dose of 100 mg/kg was administered perorally for 4 weeks and its biochemical and hepatic effects on rats with NASH were analyzed. Serum levels of glucose, triglycerides, cholesterol, and liver enzymes were quantified. Histological changes were evaluated on slices stained with H&E, trichromic and PAS. Glycogen content was measured in liver samples. α-SMA and SREBP-1 immunopositive cells were identified in liver tissue. Results NASH was characterized by elevated triglycerides, elevated liver damage enzymes, and the presence of necrosis, inflammation, steatosis, and fibrosis. Significant amounts of glycogen were found, along with α-SMA positive cells in fibrosis areas. The over-expression of SREBP-1 in cytoplasm and nuclei was evident. Animals with NASH treated with β-HPC showed a significant reduction in inflammation, necrosis, and glycogen content in the liver. A reduction in α-SMA and SREBP-1 immunopositive cells correlated with a significant reduction in the degree of fibrosis and steatosis found in liver tissue. β-HPC reduced the levels of ALP and GGT, and significantly reduced triglyceride levels. Animals treated with β-HPC did not show any alterations in liver enzyme function. Conclusions Our research shows that β-HPC can improve liver function and morphology in the case of NASH induced in rats, suggesting β-HPC could be potentially used in the treatment of NASH.

Our aim was to study the effect of the ethanol on viability, proliferation and EFGR expression in the hepatic cells (WRL68). For the study, we used the WRL‐68 cells (ATCC: CL‐48) cultured in MEM supplemented with 10 % fetal bovine serum, 2mol/L glutamine, 1% non essential amino acid. Cells were kept at 37°C, under an atmosphere 95% of air and 5% CO2. We treated hepatic cells with ethanol at concentrations of 5, 12, 25, 50, 100, 200, 400 and 800 mg/dL. The viability and cell cycle was analyzed using a MTT assay. Cell cycle and apoptosis were analyzed at different times (cell cycle at 9, 12, 15 and 24 h, and apoptosis at 24 h) by flow cytometry. EGFR expression was analyzed by western‐blot. The ethanol did not modify the viability and cell proliferation. Ethanol induced changes on S phase at 9–15h but did not alter the Go/G1 and G2+M phases in WRL68 cells. Ethanol induces death cell, we observed the presence of apoptotic (40%) and necrotic cells (10%). We observed the EGFR expression in the hepatic cell line WRL68. However, no changes in EGFR expression were observed in cells treated with ethanol. In conclusion, ethanol induces apoptosis, necrosis and changes in cell cycle, but not changes in EGFR expression were observed in hepatic cells WRL‐68.

Our aim was to study the effect of genistein on EGFR phosphorylation on liver rats with acute damage induces by administration of carbon tetrachloride (CCl4). For this study, we used male Wistar rats which were randomly divided into four groups: control, genistein 5mg/kg oral (2 weeks), acute damage CCl4 4mg/kg i.p. (4 weeks) and acute damage+genistein. Rats were sacrificed and liver were collected for determinate EGFR expression and tyrosine phosphorylation by western blot and immunoprecipitation. EGFR expression was elevated in groups with acute liver damage and acute damage+genistein. On the other hand when we analyzed EGFR phosphorylation we founded a reduction on Tyr845 EGFR phosphorylation in group with liver damage, nevertheless the genistein increased this phosphorylation 5 fold compared with CCl4 group, on Tyr1045 the phosphorylation was to decrease in acute damage and acute damage+genistein group. On Tyr992 and Tyr1068 EGFR phosphorylation genistein and acute damage not showed modification. In conclusion, the genistein can be involved in process like proliferation, apoptosis and EGFR degradation in the liver during acute damage, because it modifies specific tyrosine phosphorylation in EGFR.

The epidermal growth factor receptor (EGFR) plays an important role on hepatic protection in acute and chronic liver injury. The aim of this study was to investigate the role of genistein on EGFR expression, phosphorylation and signaling pathways in experimental subacute liver damage induced by carbon tetrachloride (CCl4). We used male Wistar rats that were randomly divided into four groups: (1) Control; (2) Genistein 5 mg/kg per oral; (3) Subacute liver damage induced by CCl4 4 mg/kg subcutaneously; and (4) Animals received CCl4 and genistein at the dosage indicated. The effect of genistein on EGFR expression, phosphorylation and signaling pathways were investigated by western blot and densitometric analyses. Histological changes were evaluated on slices stained with Hematoxylin-Eosin and Masson´s trichromic, as well as an immunohistochemical analysis for proliferating cell nuclear antigen (PCNA). Additionally, pro-inflammatory cytokines and liver enzymes were quantified. Our study showed that genistein increased EGFR expression, EGFR-specific tyrosine residues phosphorylation (pY1068-EGFR and pY84-EGFR), signal transducer and activator of transcription phosphorylation (pSTAT5), protein kinase B phosphorylation (pAKT) and PCNA in animals with CCl4-induced subacute liver damage. It was found a significant reduction of pro-inflammatory cytokines in serum from animals with subacute liver damage treated with genistein. Those effects were reflected in an improvement in the architecture and liver function. In conclusion, genistein can induce a transactivation of EGFR leading to downstream cell signaling pathways as early events associated with regeneration and hepatoprotection following subacute liver damage.

Our aim was to compare analogue A21 vs amphotericin B (AmB) in a mouse model of candidiasis. For the study, we used male Balb/c mice, which were randomly divided into six groups: control, candidiasis, AmB Indofine 4mg/kg, AmB Abelcet®, analogue A21 4 mg/kg and analogue A21 12 mg/kg. Candidiasis model was carried out by inoculated 5×107UFC/mL of Candida albicans 10231 yest P.O., 23 days after the treatment was given for the group along two weeks. Mice were sacrificed and liver, small intestine, lung and kidney were collected and then sectioned and fixed in phosphate‐buffered 4% formaldehyde for histological analysis. Samples were stained with hematoxylin‐eosin. Tissues with candidiasis showed alterations in their structures, in kidney, the number of glomeruli and tubules were decreased, while in the liver showed necrosis and apoptosis areas, also in intestine and lung showed alteration on their architecture. The analogue A21 at two concentrations impoved the structure of lungs and intestine, in kidney improved the structure and increase the number of glomeruli as AmB, in liver improved the structure at 12mg/kg. In conclusion, the analogue A21 improves the structure of tissues damaged by candidiasis like amphotericinB.