Journal: bioRxiv
Article Title: An LSEC-focused computational drug repurposing platform for liver fibrosis: Identification of vorinostat and other LSEC-protective candidates
doi: 10.64898/2026.05.23.727430
Figure Lengend Snippet: (A) Summary of hepatocyte-derived factors in intercellular crosstalk during NAFLD pathogenesis. (B) Asah1 floxed ( Asah1 fl/fl /WT) mice and Asah1 fl/fl / Alb Cre mice (hepatocyte-specific deletion of Asah1 ) were fed with PD for 20 weeks. Hepatic and plasma HMGB1 levels were measured by ELISA. Hepatic HMGB1 level was normalized to total protein. (C) HepG2-hepatocytes were transfected with siNC or si ASAH1 for 48 hours, followed by treatment with vehicle or a lipid mixture consisting of free FAs (300 μM; OA: PA = 2:1) and 7K (40 μM) for 24 hours. Cell lysate and supernatant HMGB1 levels were quantified by ELISA. (D) Transwell co-culture schematic. HepG2 hepatocytes were cultured in the upper chamber, transfected with siNC or siASAH1 for 48 hours. Primary LSECs were cultured separately in the lower chamber. After transfection, co-culture was initiated with FA + 7K treatment in the upper chamber and vehicle or glycyrrhizin (60 μM, HMGB1 inhibitor) in the lower chamber for 24 hours. (E) qPCR quantification (normalized to HPRT1 ) and (F) heatmap of LSEC marker expression. Mean ± SEM; n = 4. * p < 0.05, ** p < 0.01. Abbreviations: HMGB1, high mobility group box 1; HSCs, hepatic stellate cells; RAGE, receptor for advanced glycation end products; pMEK1/2, phosphorylated mitogen-activated protein kinase kinase 1/2; pERK1/2, phosphorylated extracellular signal-regulated kinase 1/2; CCl₄, carbon tetrachloride; TAA, thioacetamide; BDL, bile duct ligation; ASH, alcoholic steatohepatitis; MCD, methionine-choline-deficient; NASH, nonalcoholic steatohepatitis; Hmgb1ΔHepΔMye, hepatocyte and myeloid cell-specific HMGB1 knockout; SHH, Sonic hedgehog; IHH, Indian hedgehog; TAZ, transcriptional co-activator with PDZ-binding motif (WWTR1); TEAD, TEA domain transcription factor; OPN, osteopontin; CEBPA, CCAAT/enhancer-binding protein alpha; EVs, extracellular vesicles; PPAR-γ, peroxisome proliferator-activated receptor gamma; α-SMA, alpha-smooth muscle actin; TIMP-2, tissue inhibitor of metalloproteinase 2; VEGF-A, vascular endothelial growth factor A; PI3K, phosphoinositide 3-kinase; Akt, protein kinase B; VEGFR-2, vascular endothelial growth factor receptor 2; mAb, monoclonal antibody; VWF, von Willebrand factor; eNOS, endothelial nitric oxide synthase; ITGβ1, integrin beta 1; ITGα9β1, integrin alpha 9 beta 1; VCAM-1, vascular cell adhesion molecule 1; MoMF, monocyte-derived macrophage; CCL2, C-C motif chemokine ligand 2; RBPJ, recombination signal binding protein for immunoglobulin kappa J region; MCP-1, monocyte chemoattractant protein-1; CCR2, C-C motif chemokine receptor 2; IRE1α, inositol-requiring enzyme 1 alpha; S1P, sphingosine-1-phosphate; S1PR1, sphingosine-1-phosphate receptor 1; FOXO1, forkhead box O1; CXCL10, C-X-C motif chemokine ligand 10; MLK3, mixed lineage kinase 3; TRAIL, TNF-related apoptosis-inducing ligand; DR5, death receptor 5; RIP1, receptor-interacting protein 1; mtDNA, mitochondrial DNA; TLR9, Toll-like receptor 9; FA, fatty acid; 7K, 7-ketocholesterol; siNC, negative control siRNA; siASAH1, ASAH1 siRNA; Gly, Glycyrrhizin; HPRT1, hypoxanthine phosphoribosyltransferase; NLRP3, NLR family pyrin domain containing 3; CASP1, caspase-1; ICAM1, intercellular adhesion molecule 1; GSDMD, gasdermin D; LYVE1, lymphatic vessel endothelial hyaluronan receptor 1; KDR, kinase insert domain receptor; PLVAP, plasmalemma vesicle-associated protein; COL4A1, collagen type IV alpha 1 chain; COL4A2, collagen type IV alpha 2 chain; TGF-β, transforming growth factor beta; MMP2, matrix metallopeptidase 2; MMP9, matrix metallopeptidase 9; NOS3, nitric oxide synthase 3; ET-1, endothelin-1.
Article Snippet: Human hepatocellular carcinoma cell line HepG2 was purchased from ATCC and maintained in Eagle’s minimum essential medium (EMEM) with 10% FBS.
Techniques: Derivative Assay, Clinical Proteomics, Enzyme-linked Immunosorbent Assay, Transfection, Co-Culture Assay, Cell Culture, Marker, Expressing, Ligation, Knock-Out, Binding Assay, Negative Control