Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: Gel electrophoresis separation of PCR products. (A-D) PCR products of genomic DNA samples by using HeLa gDNA detection primer set. (E-H) products of genomic DNA samples by using Human gDNA control primer set. (A and E) HeLa positive control PCR template, (B and F) HeLa cells, (C and G) L02 cells mixed with 1% HeLa cells and (D and H) L02 cells. HLD, HeLa gDNA detection primer set; HGC, Human gDNA control primer set.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Nucleic Acid Electrophoresis, Control, Positive Control

Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: PAE increases LPS decreased cell viability of L02 cells. (A) The injury effect of LPS on cell viability of L02 cells. Cells were incubated with different concentration of LPS (0.05, 0.5, 5.0 or 10.0 µg/ml) for 12 h. (B) The effect of PAE alone on cell viability of L02 cells. Cells were incubated with different concentration of PAE (0.1, 1.0, 10.0 or 100.0 µg/ml) for 12 h. (C) The protective effect of PAE on LPS damaged L02 cells. Cells were incubated with different concentration of PAE (0.1, 1.0 or 10.0 µg/ml) prior 1 h and then co-incubated with LPS (10 µg/ml) for 12 h. Cell viability was assessed by CCK-8. Data represent mean ± standard error of the mean of six independent experiments. *P<0.05 or **P<0.01 vs. control or LPS. PAE, paeonol; LPS, lipopolysaccharide.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Incubation, Concentration Assay, CCK-8 Assay, Control

Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: PAE inhibits LPS increased ALT and AST of L02 cells. Cells were incubated with LPS at 10.0 µg/ml for 12 h or cells were incubated with PAE at 10.0 µg/ml prior 1 h and then co-incubated with LPS (10 µg/ml) for 12 h. The activities of (A) ALT and (B) AST were assayed according to the manufacturer's instructions. Data represent mean ± standard error of the mean of six independent experiments. *P<0.05 or **P<0.01 vs. control. PAE, paeonol; LPS, lipopolysaccharide; ALT, alanine aminotransferase; AST, aspartate transaminase.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Incubation, Control

Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: PAE inhibits LPS induced the apoptosis of L02 cells. Cells were incubated with LPS at 10.0 µg/ml for 12 h or cells were incubated with PAE at 10.0 µg/ml prior 1 h and then co-incubated with LPS (10 µg/ml) for 12 h. Cell apoptosis was observed with laser confocal microscopy (scale bar=75 µm). The population was separated into three groups: Live cells with a low level of fluorescence, apoptotic cells in the earlier period with green fluorescence and necrotic and advanced stage apoptotic cells with both red and green fluorescence. PAE, paeonol; LPS, lipopolysaccharide.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Incubation, Confocal Microscopy, Fluorescence

Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: PAE decreases LPS induced ROS generation of L02 cells (bar=75µm). Cells were incubated with LPS at 10.0 µg/ml for 12 h or cells were incubated with PAE at 10.0 µg/ml prior 1 h and then co-incubated with LPS (10 mg/ml) for 4 h. Intracellular ROS was observed with laser confocal microscopy. The population was separated into two groups: Left panels were cells observed by phase contrast and right panels were cells observed by fluorescence microscopy. PAE, paeonol; LPS, lipopolysaccharide; ROS, reactive oxygen species; DCF, dichlorofluorescein.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Incubation, Confocal Microscopy, Fluorescence, Microscopy

Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: PAE inhibits LPS increased MDA and increases LPS decreased SOD level of L02 cells. Cells were incubated with LPS at 10.0 µg/ml for 12 h or cells were incubated with PAE at 10.0 µg/ml prior 1 h and then co-incubated with LPS (10 µg/ml) for 12 h. (A) MDA and (B) SOD were assayed by respective kits. Data represent mean ± standard error of the mean of six independent experiments. *P<0.05 or **P<0.01 vs. control. PAE, paeonol; LPS, lipopolysaccharide; MDA, malondialdehyde; SOD, superoxide dismutase.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Incubation, Control

Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: PAE increases LPS-decreased mitochondrial transmembrane potential in L02 cells (bar=75 µm). Cells were incubated with LPS at 10.0 µg/ml for 12 h or cells were incubated with PAE at 10.0 µg/ml prior 1 h and then co-incubated with LPS (µg/ml) for 12 h. The mitochondrial membrane potential was measured by laser confocal microscopy. The population was separated into four groups: Left panels were cells observed by bright light and right panels were the merged field of red (for JC-1 aggregates) and green color (for JC-1 monomers). PAE, paeonol; LPS, lipopolysaccharide.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Incubation, Membrane, Confocal Microscopy

Journal: Molecular Medicine Reports

Article Title: Paeonol alleviates lipopolysaccharide-induced hepatocytes injury through alteration of mitochondrial function and NF-κB translocation

doi: 10.3892/mmr.2021.12419

Figure Lengend Snippet: PAE inhibits LPS-promoted nuclear translocation of NF-κB (scale bar=75 µm). Cells were incubated with LPS at 10.0 µg/ml for 12 h or cells were incubated with PAE at 10.0 µg/ml prior 1 h and then co-incubated with LPS (10 µg/ml) for 12 h. The NF-κB level in the nuclei of L02 cells was detected by immunofluorescence staining. The right panels were the merged field of red (for NF-κB subunit p 65) and blue color (for cell nucleus). PAE, paeonol; LPS, lipopolysaccharide.

Article Snippet: L02 human normal hepatocytes were purchased from BioVector NTCC Inc. and the cells were cultured in a 5% CO 2 atmosphere at 37°C.

Techniques: Translocation Assay, Incubation, Immunofluorescence, Staining

Journal: Scientific Reports

Article Title: Sodium aescinate-induced hepatotoxicity via ATF4/GSH/GPX4 axis-mediated ferroptosis

doi: 10.1038/s41598-024-79723-2

Figure Lengend Snippet: SA induces ferroptosis in mouse livers. Mice were intraperitoneally (i.p.) injected with vehicle, SA (3.6, 7.2 mg/kg), Fer-1 (5 mg/kg/d) or SA (3.6 mg/kg/d) + Fer-1 (5 mg/kg/d). After 10 d, the relative liver weight index was measured ( A ). Serum AST and ALT in each group were measured ( B ). Images of H&E staining (HE) of the liver Sect. (200 × magnification): ① edema of hepatocytes, ② compensatory hypertrophy of liver ( C ). Hepatic MDA ( D ), GSH ( E ) and Fe 2+ content ( F ) were assessed. Values are expressed as the mean ± SD ( n = 6). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group.

Article Snippet: Human normal hepatocytes (L02 cells) were obtained from the American Type Culture Collection.

Techniques: Injection, Staining, Control

Journal: Scientific Reports

Article Title: Sodium aescinate-induced hepatotoxicity via ATF4/GSH/GPX4 axis-mediated ferroptosis

doi: 10.1038/s41598-024-79723-2

Figure Lengend Snippet: ATF4 is involved in SA-induced hepatocyte ferroptosis. After pretreated with ATF4 si-RNA, protein levels of N-ATF4, t-ATF4, xCT, CTH and GPX4 in L02 cells were determined ( A ). L02 cells were transfected with ATF4 expression plasmid (ATF4-OE) or the empty vector or ATF4 si-RNA or si-con. Cells were then exposed to SA, and cell viability ( B ), LDH leakage ( C ), the levels of cysteine ( D ), GSH ( E ), MDA and Fe 2+ ( F ), were detected. Intracellular ROS was assessed by DCFH-DA ( G , I ). Intracellular lipid peroxidation was assessed by BODIPY 581/591 C11 dye ( H , J ). After transfected with xCT expression plasmid (xCT-OE) or CTH expression plasmid (CTH-OE), the expression levels of xCT or CTH were determined ( K , L ). L02 cells were transfected with ATF4 si-RNA or not and with xCT expression plasmid (xCT-OE) or CTH expression plasmid (CTH-OE). Cells were then exposed to SA, and the levels of cysteine ( M ), GSH ( N ), MDA ( O ) and cell viability ( P ) were detected. Values are expressed as the mean ± SD ( n = 3). * p < 0.05, ** p < 0.01, *** p < 0.001 vs. control group. # p < 0.05 vs. SA group. & p < 0.05 vs. SA + ATF4 si-RNA group.

Article Snippet: Human normal hepatocytes (L02 cells) were obtained from the American Type Culture Collection.

Techniques: Transfection, Expressing, Plasmid Preparation, Control