human normal hepatocytes thle2  (ATCC)

Journal: Frontiers in Pharmacology

Article Title: Ultrafine garlic powder alleviates non-alcoholic steatohepatitis by inhibiting hepatocyte ferroptosis and modulating ERK-dependent oxidative stress

doi: 10.3389/fphar.2025.1711917

Figure Lengend Snippet: UGP ameliorates palmitatic acid-induced lipotoxic hepatocyte injury. Effects of palmitatic acid stimulation and Fer-1, UGP and TGP treatment of on human normal hepatocyte THLE2 cell viability was examined using CCK8 assay (A) and LDH assay (B) (n = 4). (C) Effects of palmitatic acid stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell damage was determined by ALT assay (n = 4). (D) Cell death assay was performed using Annexin V/PI staining (n = 4). Annexin V + PI − , Annexin V + PI − , and Annexin V + PI + cells were considered dead cells. Representative fluorescent images and quantification of DCFDA (2′,7′-dichlorofluorescein diacetate) (E) and Mito-Tracker Red CMXRos (F) staining for ROS and mitochondrial membrane potential detection (scale bar = 100 μm, n = 4). (G) Flow cytometry analysis of lipid peroxidation by Liperfluo staining on THLE2 cells. (H) Representative immunofluorescent images of ACTA2 +ve and COL1A1 +ve cells (scale bar = 100 μm). Data are shown as box-and whisker with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers), one-way ANOVA with Tukey’s correction. (ALT: Alanine transaminase; BSA:Bovine serum albumin; Fer-1: Ferrostatin-1; PAL: Palmitic acid; ROS: Reactive oxygen species; TGP: Traditional garlic powder; UGP:Ultrafine garlic powder).

Article Snippet: Human normal hepatocytes (THLE2) and hepatic stellate cells (HSCs) (LX-2) were purchased from the American Type Culture Collection (ATCC) and cultured in BEGM kit medium and RPMI medium supplemented with 10% FBS, respectively.

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, Staining, Membrane, Flow Cytometry, Whisker Assay

Journal: Frontiers in Pharmacology

Article Title: Ultrafine garlic powder alleviates non-alcoholic steatohepatitis by inhibiting hepatocyte ferroptosis and modulating ERK-dependent oxidative stress

doi: 10.3389/fphar.2025.1711917

Figure Lengend Snippet: UGP significantly ameliorates erastin-induced hepatocyte ferroptosis. Effects of erastin stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell viability was examined using CCK8 assay (A) and LDH assay (B) (n = 4). (C) Effects of erastin stimulation and Fer-1, UGP and TGP treatment of on THLE2 cell damage was determined by ALT assay (n = 4). (D) Cell death assay was performed using Annexin V/PI staining (n = 4). Annexin V + PI − , Annexin V + PI − , and Annexin V + PI + cells were considered dead cells. (E) Flow cytometry analysis of cellular Fe 2+ levels by FerroOrange staining on THLE2 cells. Representative fluorescent images and quantification of DCFDA (2′,7′-dichlorofluorescein diacetate) (F) and Mito-Tracker Red CMXRos (G) staining for ROS and mitochondrial membrane potential detection (scale bar = 100 μm, n = 4). (H) Flow cytometry analysis of lipid peroxidation by Liperfluo staining on THLE2 cells. Data are shown as box-and whisker with median (middle line), 25th-75th percentiles (box), and min-max values (whiskers), one-way ANOVA with Tukey’s correction. (ALT: Alanine transaminase; ERA: Erastin; Fer-1: Ferrostatin-1; ROS: Reactive oxygen species; TGP: Traditional garlic powder; UGP: Ultrafine garlic powder).

Article Snippet: Human normal hepatocytes (THLE2) and hepatic stellate cells (HSCs) (LX-2) were purchased from the American Type Culture Collection (ATCC) and cultured in BEGM kit medium and RPMI medium supplemented with 10% FBS, respectively.

Techniques: CCK-8 Assay, Lactate Dehydrogenase Assay, Staining, Flow Cytometry, Membrane, Whisker Assay