Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p was decreased in the livers of MAFLD mice and FFA-induced HepG2 cells. (A) MiR-300-3p was decreased in the livers of MAFLD mice. (B) MiR-300-3p was downregulated in the FFA-induced HepG2 cells. (C) MiR-300-3p was downregulated in HepG2 cells successfully. (D) MiR-300-3p was overexpressed in HepG2 cells successfully. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01. NC, Healthy control; OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Control, Over Expression, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: Downregulation of miR-300-3p promotes FFA-induced lipid accumulation and hepatic inflammation in HepG2 cells. (A) The experimental flow of the MAFLD cell model construction and miR-300-3p mimics, miR-300-3p inhibitors and as well as corresponding scrambled controls transfection in HepG2 cells. (B,C) Intercellular TG contents in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (D) Oil Red O staining (400×) and the relative areas of lipid droplets in miR-300-3p-inhibited and -overexpressing in HepG2 cells. (E,F) mRNA levels of FASN, SREBP-1c, IL-6, IL-2 and TNF-αin miR-300-3p -inhibited and -overexpressing in HepG2 cells. (G) Protein levels of FASN, SREBP-1c and TNF-α in miR-300-3p -inhibited and -overexpressing in HepG2 cells. Data in (B,C) and (E–G) are means±SDs (n = 3). *P < 0.05, **P < 0.01, *** P < 0.001, **** P < 0.0001. OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Transfection, Staining, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: Downregulation of miR-300-3p induces apoptosis in HepG2 cells to promote it. (A,B) The apoptosis rate in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (C) mRNA levels of Bax, Bcl-2 and Caspase-3 in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (D) Protein levels of Bax, Bcl-2 and Caspase-3 in miR-300-3p -overexpressing and -inhibited in HepG2 cells. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, over-expression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p regulates STX17 in HepG2 cells. (A,B) GO enrichment analysis (Biological Process category) and KEGG enrichment analysis diagrams of sixteen predicted target genes of miR-300-3p. (C) The predicted binding sites between miR-300-3p and STX17 were putatively identified via the TargetScan database. (D) Dual-luciferase reporter assay (DLRA) in HepG2 cells validated that STX17 is a direct target gene of miR-300-3p. (E,F) The mRNA and protein expression levels of STX17 in HepG2 cells with miR-300-3p inhibition or overexpression. Data in (A–D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Binding Assay, Luciferase, Reporter Assay, Expressing, Inhibition, Over Expression, Control

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: P62 and LC3-II in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (A) The protein levels of P62 and LC3-II in miR-300-3p -inhibited and -overexpressing in HepG2 cells. (B) P62 and LC3-II in miR-300-3p -inhibited and -overexpressing by Immunofluorescence assay. Data in (A-D) are means±SDs (n = 3). *P < 0.05, **P < 0.01,***P < 0.001,****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Immunofluorescence, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: MiR-300-3p promoted autophagy by regulating STX17, reduces lipid accumulation, inflammatory response, and decreases hepatocyte apoptosis. (A) STX17 was inhibited or overexpressed in HepG2 cells successfully. (B) Protein levels of FASN, SREBP-1c and TNF-αin STX17 -inhibited and -overexpressing in HepG2 cells. (C,D) Intercellular TG contents in STX17-inhibited and -overexpressing in HepG2 cells. (E) Oil Red O staining (400×) and relative areas of lipid droplets in STX17-inhibited and -STX17 in HepG2 cells. (F) Protein levels of Bax, Capase-3 and Bcl-2 in STX17 -inhibited and -overexpressing in HepG2 cells. Data in (A–D) and (F) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, miR-300-3p overexpression control; IC,miR-300-3p inhibition control; Control, STX17-overexpression control or STX17-inhibition control; Si-STX17, STX17-inhibition; Over-STX17, STX17-overexpression.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Staining, Over Expression, Control, Inhibition

Journal: Frontiers in Pharmacology

Article Title: Downregulation of microRNA-300-3p promotes steatosis-to-MASH progression by regulating STX17

doi: 10.3389/fphar.2026.1804415

Figure Lengend Snippet: P62, LC3-II in STX17-inhibited and -overexpressing in HepG2 cells. (A) Protein levels of P62, LC3-II in STX17-inhibited and -overexpressing in HepG2 cells. (B) P62 and LC3-II in STX17-inhibited and -overexpressing by Immunofluorescence assay. Data in (A-D) are means±SDs (n = 3). *P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001. OC, overexpression control; IC, inhibition control.

Article Snippet: The human hepatocyte cell line HepG2 was procured from the American Type Culture Collection (ATCC, United States) and cultivated in a humidified incubator maintained at 37 °C with a 5% CO 2 atmosphere.

Techniques: Immunofluorescence, Over Expression, Control, Inhibition

Journal: International Journal of Biological Sciences

Article Title: Cholesterol Overload Drives Hepatic Steatosis by Inhibiting OGT-dependent PPARα O-GlcNAcylation and Transactivation

doi: 10.7150/ijbs.135054

Figure Lengend Snippet: Impaired mitochondrial fatty acid β-oxidation is a key metabolic defect contributing to cholesterol-induced hepatocellular fat accumulation. ( A, B ) Plasma glycerol and TAG levels in chow-fed mice supplemented with and without 2% cholesterol (w/w) for 4 weeks. ( C-E ) Extracellular TAG concentrations in the culture medium of AML12, HepaRG, and HepG2 cells following cholesterol. ( F, G ) Extracellular TAG concentrations in the culture medium of AML12 and HepG2 cells following LDL supplementation. ( H, I ) Heatmap visualization of representative lipid-related pathways in AML12 cells and in vivo . ( J, K ) qRT-PCR validation of mitochondrial fatty acid β-oxidation ( Pparα , Cpt1α , Acox1 , Hmgcs2 , and Cyp7a1 ) in vitro and in vivo . ( L-N ) Unaltered Srebp-1c pathway activation in AML12 cells following cholesterol exposure and in chow-fed mice supplemented with and without 2% cholesterol for 4 weeks, tested by Western-blot and qRT-PCR. ( O, P ) Fluorescence-based FAOBlue™ assay of fatty acid β-oxidation capacity (FAO) in AML12 and HepG2 cells after cholesterol loading (4 μM, 16 h). (40× objective). Chol, cholesterol; MβCD-chol, MβCD-cholesterol; LDL, low-density lipoprotein. * P < 0.05 and ** P < 0.01 represent statistical significance.

Article Snippet: Murine AML12 and human HepG2 hepatocyte cell lines were obtained from the American Type Culture Collection (ATCC, VA, USA).

Techniques: Clinical Proteomics, In Vivo, Quantitative RT-PCR, Biomarker Discovery, In Vitro, Activation Assay, Western Blot, Fluorescence

Journal: International Journal of Biological Sciences

Article Title: Cholesterol Overload Drives Hepatic Steatosis by Inhibiting OGT-dependent PPARα O-GlcNAcylation and Transactivation

doi: 10.7150/ijbs.135054

Figure Lengend Snippet: OGT-mediated O-GlcNAcylation is essential for hepatic PPARα transactivation. AML12 cells were treated with OSMI-1. ( A ) qRT-PCR analysis of Ogt , Pparα , Hmgcs2 , and Cpt1a mRNA levels. AML12 and HepG2 cells were treated with OSMI-1, either alone or in combination with TMG. ( B, C ) Western blot analyses of PPARα protein. ( D, E ) qRT-PCR and ELISA analyses showing that Ogt silencing reduced Pparα , Hmgcs2 , and Cpt1α expression and decreased PPARα transcriptional activity in AML12 cells. ( F, G ) FAOBlue TM assay in AML12 and HepG2 cells. PPARα O-GlcNAcylation was examined by co-immunoprecipitation. ( H ) OGT association with PPARα (IB: OGT) and O-GlcNAc modification of PPARα (IB: O-GlcNAc; arrow). Right: effects of cholesterol treatment or si Ogt on PPARα O-GlcNAcylation. ( I, J ) qRT-PCR analysis of hepatic Pparα , Hmgcs2 , Cpt1α , Acox1 , and Cyp7a1 mRNA levels in liver-specific OGT- and OGA-deficient mice. O-GlcNAc, O-GlcNAcylation. * P < 0.05 and ** P < 0.01 represent statistical significance.

Article Snippet: Murine AML12 and human HepG2 hepatocyte cell lines were obtained from the American Type Culture Collection (ATCC, VA, USA).

Techniques: Quantitative RT-PCR, Western Blot, Enzyme-linked Immunosorbent Assay, Expressing, Activity Assay, Immunoprecipitation, Modification