Journal: Hepatology Communications

Article Title: Hydrogen peroxide damage to rat liver sinusoidal endothelial cells is prevented by n -acetyl-cysteine but not GSH

doi: 10.1097/HC9.0000000000000617

Figure Lengend Snippet: Influence of hydrogen peroxide–induced ROS on rat LSEC viability. Functional viability/reducing potential of the cell was assessed using resazurin/resorufin assay (A) and structural viability/cell integrity was studied with LDH-release assay (B). Measurements were conducted under continuous treatment with hydrogen peroxide on selected time points (0.5–5 h). The cell viability was calculated and normalized to the untreated control for resazurin assay and to the total LDH after cell lysis for LDH-release assay. Average data from 3 independent experiments/bioreplicates ±SD are presented. Statistical analyses below graphs show p values for trend analysis in time and concentration dependency. ANOVA for different times and concentrations compared to control values of nontreated cells; * p<0.05, ** p<0.01, *** p<0.001, ns non-significant. Abbreviations: LDH, lactase dehydrogenase; ROS, reactive oxygen species.

Article Snippet: LSECs were seeded on 48-well plates (300,000 cells/well), and a luminescence lactase dehydrogenase (LDH) detection kit (LDH-Glo, Promega) was used following the manufacturer’s instructions to assess cell viability.

Techniques: Functional Assay, Lactate Dehydrogenase Assay, Control, Resazurin Assay, Lysis, Concentration Assay

Journal: Hepatology Communications

Article Title: Hydrogen peroxide damage to rat liver sinusoidal endothelial cells is prevented by n -acetyl-cysteine but not GSH

doi: 10.1097/HC9.0000000000000617

Figure Lengend Snippet: The effect of ROS-inducing versus ROS-depleting agents on rat LSEC viability, functions, and morphology. Cells were treated with ROS-inducing agents, hydrogen peroxide, or ROS-depleting agents: GSH or NAC. For cotreatments, samples were simultaneously treated with both hydrogen peroxide and GSH/NAC at the starting point of the assays. For pretreatments, samples were treated with GSH/NAC for 30 minutes, then rinsed with fresh media and treated with H 2 O 2 at the starting point of the assays. The functional (A) and structural (C) viability were studied using resazurin and LDH release assays, respectively; (B) intracellular ROS levels were assessed using a fluorescence-based ROS detection assay after 1 hour of treatment and (D) endocytic activity was measured with radiolabeled FSA-based scavenging assay during 2 hours of treatment. (E) LSEC morphology was quantitatively assessed using SEM images. Colors represent data from separate bioreplicates. Abbreviations: FSA, formaldehyde-treated serum albumin; GSH, glutathione; LDH, lactase dehydrogenase; NAC, n -acetyl cysteine; ROS, reactive oxygen species; SEM, scanning electron microscopy.

Article Snippet: LSECs were seeded on 48-well plates (300,000 cells/well), and a luminescence lactase dehydrogenase (LDH) detection kit (LDH-Glo, Promega) was used following the manufacturer’s instructions to assess cell viability.

Techniques: Functional Assay, Fluorescence, Detection Assay, Activity Assay, Electron Microscopy