Journal: Frontiers in Pharmacology

Article Title: Luteolin protects against cholestasis by suppressing hepatocyte apoptosis through direct PI3K binding and AKT activation

doi: 10.3389/fphar.2026.1930755

Figure Lengend Snippet: Primers used in this study.

Article Snippet: The PI3K protein (P27-102CH, Sino Biological, Beijing, China) was formulated into a 10 mM sodium acetate buffer (pH 4.0) and then immobilized onto the detection channel of the M5 chip.

Techniques:

Journal: Frontiers in Pharmacology

Article Title: Luteolin protects against cholestasis by suppressing hepatocyte apoptosis through direct PI3K binding and AKT activation

doi: 10.3389/fphar.2026.1930755

Figure Lengend Snippet: Network pharmacology predicts therapeutic mechanism of Lut against cholestasis. (A) Venn diagram showing 142 intersecting targets between Lut and cholestasis. (B) Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis of the intersecting targets. Bubble size indicates the number of enriched genes, and color gradient represents the significance level ( p -value); the x-axis represents rich factor; the y-axis lists enriched pathways. (C) PPI network of the 142 intersecting targets. (D) Top 10 core targets of the PPI network.

Article Snippet: The PI3K protein (P27-102CH, Sino Biological, Beijing, China) was formulated into a 10 mM sodium acetate buffer (pH 4.0) and then immobilized onto the detection channel of the M5 chip.

Techniques: Analysis

Journal: Frontiers in Pharmacology

Article Title: Luteolin protects against cholestasis by suppressing hepatocyte apoptosis through direct PI3K binding and AKT activation

doi: 10.3389/fphar.2026.1930755

Figure Lengend Snippet: Binding analysis of Lut to PI3K via molecular docking and SPR. (A) The steady-state fit curve and dynamic fit curve of Lut and PI3K via SPR. (B) The predicted binding mode of Lut within the kinase domain of PI3K (PDB ID: 5DXT). The full-length PI3K catalytic subunit structure is shown as a green ribbon, Lut is depicted as yellow stick structures. The enlarged view shows the interaction between Lut and key residues within the ATP-binding pocket.

Article Snippet: The PI3K protein (P27-102CH, Sino Biological, Beijing, China) was formulated into a 10 mM sodium acetate buffer (pH 4.0) and then immobilized onto the detection channel of the M5 chip.

Techniques: Binding Assay, Analysis, Docking Assay

Journal: Frontiers in Pharmacology

Article Title: Luteolin protects against cholestasis by suppressing hepatocyte apoptosis through direct PI3K binding and AKT activation

doi: 10.3389/fphar.2026.1930755

Figure Lengend Snippet: Lut activates PI3K/AKT signaling and regulates apoptosis-related factors in vivo . (A–E) The relative mRNA expression levels of PI3K (A) , AKT (B) , BCL2 (C) , BAX (D) , and Caspase 3 (E) in liver tissues were determined by RT-qPCR. (F,G) Representative Western blot images showing the protein expression of PI3K, p-PI3K, AKT, p-AKT, BCL2 (F) , and BAX, Caspase 3, cleaved-Caspase 3 (cl-Caspase 3) (G) . GAPDH was used as the loading control for normalization of each corresponding protein expression analysis. (H-L) Quantitative analysis of the Western blot results: p-PI3K/PI3K ratio (H) , p-AKT/AKT ratio (I) , relative BCL2 protein level (J) , relative BAX protein level (K) , and cl-Caspase 3/Caspase 3 ratio (L) . (M) Representative immunofluorescence images of TUNEL staining in liver tissues. Red fluorescence indicates TUNEL-positive apoptotic cells, and blue fluorescence indicates DAPI-stained nuclei (Scale bar: 200 μm). (N) Quantitative analysis of the ratio of TUNEL-positive cells. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. ANIT group. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. Lut-H group. Only statistically significant comparisons are indicated in the figures.

Article Snippet: The PI3K protein (P27-102CH, Sino Biological, Beijing, China) was formulated into a 10 mM sodium acetate buffer (pH 4.0) and then immobilized onto the detection channel of the M5 chip.

Techniques: In Vivo, Expressing, Quantitative RT-PCR, Western Blot, Control, Analysis, Immunofluorescence, TUNEL Assay, Staining, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Luteolin protects against cholestasis by suppressing hepatocyte apoptosis through direct PI3K binding and AKT activation

doi: 10.3389/fphar.2026.1930755

Figure Lengend Snippet: PI3K inhibition abolishes the protective effects of Lut in HepG2 cells. (A,B) Representative Western blot images showing the protein expression of PI3K, p-PI3K, AKT, p-AKT, BCL2 (A) , and BAX, Caspase 3, cl-Caspase 3 (B) in the presence or absence of the PI3K inhibitor LY294002. GAPDH was used as the loading control. (C–G) Quantitative analysis of the Western blot results: p-PI3K/PI3K ratio (C) , p-AKT/AKT ratio (D) , relative BCL2 protein level (E) , relative BAX protein level (F) , and cl-Caspase 3/Caspase 3 ratio (G) . (H) Representative immunofluorescence images of TUNEL staining in HepG2 cells. Red fluorescence indicates TUNEL-positive apoptotic cells, and blue fluorescence indicates DAPI-stained nuclei (Scale bar: 50 μm). (I) Quantitative analysis of the ratio of TUNEL-positive cells. Data are expressed as mean ± SD. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The PI3K protein (P27-102CH, Sino Biological, Beijing, China) was formulated into a 10 mM sodium acetate buffer (pH 4.0) and then immobilized onto the detection channel of the M5 chip.

Techniques: Inhibition, Western Blot, Expressing, Control, Analysis, Immunofluorescence, TUNEL Assay, Staining, Fluorescence

Journal: Frontiers in Pharmacology

Article Title: Luteolin protects against cholestasis by suppressing hepatocyte apoptosis through direct PI3K binding and AKT activation

doi: 10.3389/fphar.2026.1930755

Figure Lengend Snippet: Schematic diagram of the protective mechanism: Lut alleviates ANIT-induced cholestatic liver injury through PI3K/AKT-dependent inhibition of hepatocyte apoptosis.

Article Snippet: The PI3K protein (P27-102CH, Sino Biological, Beijing, China) was formulated into a 10 mM sodium acetate buffer (pH 4.0) and then immobilized onto the detection channel of the M5 chip.

Techniques: Inhibition, In Vitro, SPR Assay, Docking Assay, TUNEL Assay