Journal: Frontiers in Pharmacology

Article Title: Baicalein Exerts Neuroprotective Effects in FeCl 3 -Induced Posttraumatic Epileptic Seizures via Suppressing Ferroptosis

doi: 10.3389/fphar.2019.00638

Figure Lengend Snippet: Baicalein inhibits FeCl 3 -induced behavioral seizures. (A–C) Behavioral seizures induced by different concentrations of FeCl 3 (20, 35, and 50 mM) were presented as seizure score, number of seizures, and average seizure duration (Sec), n = 6. (D–F) Intraperitoneal injection of different doses of baicalein (50 and 100 mg/kg) 30 min prior to the injection of 50 mM FeCl 3 . The seizure score, number of seizures, and average seizure duration (Sec) were reduced significantly at different doses of baicalein, n = 6. *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: The plated cells were pretreated with 32 μM baicalein (S2268, Selleck, USA) for 2 h. Then, medium containing FAC (F5879, Sigma, USA) or erastin (S7242, Selleck, USA) at the final concentration of 300 μM and 500 nM were added and cultured for 36 and 8 h, respectively.

Techniques: Injection

Journal: Frontiers in Pharmacology

Article Title: Baicalein Exerts Neuroprotective Effects in FeCl 3 -Induced Posttraumatic Epileptic Seizures via Suppressing Ferroptosis

doi: 10.3389/fphar.2019.00638

Figure Lengend Snippet: Baicalein attenuates FAC-induced neuronal damage by inhibiting ferroptosis. (A) Cell viability was measured with CCK8 assay, in which HT22 cells were incubated with various concentrations (37.5, 75, 150, and 300 μM) of FAC for different periods of time (12–36 h). (B) HT22 cells were incubated with different concentrations (1, 2, 4, 8, 16, and 32 μM) of baicalein for 2 h, prior to incubation with 300 μM of FAC for 36 h. (C) FAC exposure caused cell shrinkage and nuclear condensation. Baicalein pretreatment reduced cell death. Scale bar: 200 μm (D) after 2 h pretreatment with 32 μM baicalein, 12.5 μM Fer-1, and 1 μM Lipo-1, lipid ROS in HT22 cells was assessed by FACS analysis of C11-BODIPY fluorescence after treatment with FAC for 36 h. (E) Representative Western blots of 4-HNE with 32 μM baicalein and ferroptosis inhibitors pretreatment for 2 h in the FAC-induced HT22 cell injury model. (F) The relative mRNA expression of PTGS2 was reduced after baicalein and ferroptosis inhibitors pretreatment. (G) Representative Western blots of GPX4 with 32 μM baicalein and ferroptosis inhibitors pretreatment for 2 h in the FAC-induced HT22 cell injury model. Data are representative of three independent experiments with similar results, *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: The plated cells were pretreated with 32 μM baicalein (S2268, Selleck, USA) for 2 h. Then, medium containing FAC (F5879, Sigma, USA) or erastin (S7242, Selleck, USA) at the final concentration of 300 μM and 500 nM were added and cultured for 36 and 8 h, respectively.

Techniques: CCK-8 Assay, Incubation, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Baicalein Exerts Neuroprotective Effects in FeCl 3 -Induced Posttraumatic Epileptic Seizures via Suppressing Ferroptosis

doi: 10.3389/fphar.2019.00638

Figure Lengend Snippet: Baicalein attenuates erastin-induced neuronal damage by inhibiting ferroptosis. (A) Cell viability was measured with CCK8 assay, in which HT22 cells were incubated with various concentrations (125, 250, 500, and 1,000 nM) of erastin for 8 h. (B) HT22 cells were incubated with different concentrations of baicalein for 2 h, prior to incubation with 500 nM erastin for 8 h. (C) Baicalein (32 μM) pretreatment could reduce cell death induced by 500 nM erastin. Scale bar: 200 μm (D) after 2 h pretreatment with 32 μM baicalein, lipid ROS in HT22 cells was assessed by FACS analysis of C11-BODIPY fluorescence after treatment with erastin for 8 h. (E) Representative Western blots of 4-HNE with 32 μM baicalein pretreatment for 2 h in the erastin-induced HT22 cell injury model. (F) The relative mRNA expression of PTGS2 was reduced after baicalein pretreatment. (G) Western blots illustrated that GPX4 was significantly diminished in the erastin-induced HT22 cell injury model and baicalein treatment reversed this phenomenon. Data are representative of three independent experiments with similar results, *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: The plated cells were pretreated with 32 μM baicalein (S2268, Selleck, USA) for 2 h. Then, medium containing FAC (F5879, Sigma, USA) or erastin (S7242, Selleck, USA) at the final concentration of 300 μM and 500 nM were added and cultured for 36 and 8 h, respectively.

Techniques: CCK-8 Assay, Incubation, Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Baicalein Exerts Neuroprotective Effects in FeCl 3 -Induced Posttraumatic Epileptic Seizures via Suppressing Ferroptosis

doi: 10.3389/fphar.2019.00638

Figure Lengend Snippet: 12/15-LOX is involved in the inhibition of ferroptosis in FAC-induced HT22 cell injury model by baicalein. (A) Immunofluorescence analysis indicated that FAC and erastin significantly increased the expression of 12/15-LOX in HT22 cells, which was reversed by baicalein and PD146176. Scale bar: 50 μm (B) CCK8 analysis showed that pretreatment with PD146176 could reverse cell damage induced by FAC. (C–D) Exposure to FAC and erastin caused HT22 cells to shrink, and 1 μM PD146176 reversed this phenomenon. Scale bar: 200 μm (E–F) pretreatment with 1 μM PD14617, 12.5 μM Fer-1, and 1 μM Lipo-1 could reverse increased lipid ROS in HT22 cells induced by FAC and erastin. Data are representative of three independent experiments with similar results, *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: The plated cells were pretreated with 32 μM baicalein (S2268, Selleck, USA) for 2 h. Then, medium containing FAC (F5879, Sigma, USA) or erastin (S7242, Selleck, USA) at the final concentration of 300 μM and 500 nM were added and cultured for 36 and 8 h, respectively.

Techniques: Inhibition, Immunofluorescence, Expressing

Journal: Frontiers in Pharmacology

Article Title: Baicalein Exerts Neuroprotective Effects in FeCl 3 -Induced Posttraumatic Epileptic Seizures via Suppressing Ferroptosis

doi: 10.3389/fphar.2019.00638

Figure Lengend Snippet: Baicalein counteracts FeCl 3 -induced epileptic seizure behavior by inhibiting ferroptosis. (A) Decreased seizure score after treatment with Lipo-1 (a specific inhibitor of ferroptosis) in the FeCl 3 -induced PTE model. (B) Western blots showing expression of 4-HNE in hippocampus tissues of FeCl 3 -induced PTE mice with different doses of baicalein and Lipo-1 pretreatment, n = 6. (C) The relative mRNA expression of PTGS2 was reduced after baicalein and Lipo-1 pretreatment, n = 6. (D) Western blots illustrated that GPX4 was significantly diminished in FeCl 3 -induced PTE mice, and baicalein and Lipo-1 treatment reversed this phenomenon (n = 6 for both groups). (E) Baicalein inhibited the increase of 12/15-LOX in hippocampus tissues from FeCl 3 -induced PTE mice, n = 6. *p < 0.05, **p < 0.01, and ***p < 0.001.

Article Snippet: The plated cells were pretreated with 32 μM baicalein (S2268, Selleck, USA) for 2 h. Then, medium containing FAC (F5879, Sigma, USA) or erastin (S7242, Selleck, USA) at the final concentration of 300 μM and 500 nM were added and cultured for 36 and 8 h, respectively.

Techniques: Western Blot, Expressing

Journal: Frontiers in Pharmacology

Article Title: Baicalein Exerts Neuroprotective Effects in FeCl 3 -Induced Posttraumatic Epileptic Seizures via Suppressing Ferroptosis

doi: 10.3389/fphar.2019.00638

Figure Lengend Snippet: The graph depicted the neuroprotective role of baicalein in FeCl 3 -induced posttraumatic epileptic seizures. Baicalein ameliorated the FeCl 3 -induced posttraumatic epileptic seizures by inhibiting 12/15-LOX-associated ferroptosis.

Article Snippet: The plated cells were pretreated with 32 μM baicalein (S2268, Selleck, USA) for 2 h. Then, medium containing FAC (F5879, Sigma, USA) or erastin (S7242, Selleck, USA) at the final concentration of 300 μM and 500 nM were added and cultured for 36 and 8 h, respectively.

Techniques:

Journal: Cell Death and Differentiation

Article Title: Resolution of inflammation and sepsis survival are improved by dietary Ω-3 fatty acids

doi: 10.1038/cdd.2017.177

Figure Lengend Snippet: Ω-3 + LEs enhance human MΦ efferocytosis and phagocytosis through 5-LOX and 12/15-LOX. Human MΦ were incubated with indicated concentrations of Ω-3 + or Ω-3 − LEs. ( a ) The rate of efferocytosis of apoptotic PMN or ZyA-labeled particles was assessed by immunofluorescence and illustrated by immunofluorescence images. ( b ) The rate of efferocytosis of fluorescently labeled E. coli bacteria was performed using a fluorescent reader. ( c ) mRNA expression of ALX/FPR2, DRV1/GPR32 and ERV/ChemR23 on human MΦ following stimulation with Ω-3 + LEs or ( d ) Ω-3 − LEs. The results are representative of 8–14 independent experiments and are expressed as the mean±S.E.M., * P <0.05; ** P <0.01; *** P <0.001, one-way ANOVA, followed by Dunnett’s multiple-comparison test. In a different experiment, ( e ) peritoneal MΦ from WT or 12/15-LOX −/− mice were incubated with Ω-3 + or Ω-3 − LEs, and the rate of phagocytosis of fluorescently labeled ZyA particles was assessed using a fluorescent plate reader. ( f ) Human MΦ were incubated with indicated concentrations of baicalein and CDC and the degree of phagocyted fluorescently labeled ZyA particles was assessed. Human MΦ were incubated with Ω-3 + or Ω-3 − LEs in the absence or presence of baicalein or CDC and the rate of MΦ clearance of ZyA particles was performed. Results are representative of 5–10 independent experiments and are expressed as the mean±S.E.M., * P <0.05; ** P <0.01; *** P <0.001, one-way ANOVA, followed by Dunnett’s or Bonferroni’s multiple-comparison test

Article Snippet: In a further experiment, human MΦ were incubated with baicalein (Santa Cruz Biotechnology) or cinnamyl-3,4dihydroxy-α-cyanocinnamate (CDC, Santa Cruz Biotechnology) and the degree of phagocytosis was assessed by using a fluorescent plate reader (Tecan, Männedorf, Switzerland).

Techniques: Incubation, Labeling, Immunofluorescence, Bacteria, Expressing, Comparison

Journal: Nutrients

Article Title: Wogonin, a Compound in Scutellaria baicalensis , Activates ATF4–FGF21 Signaling in Mouse Hepatocyte AML12 Cells

doi: 10.3390/nu14193920

Figure Lengend Snippet: Wogonin induces Atf4 and Fgf21 expression. ( A ) Wogonin increased Fgf21 expression in AML12 cells. Cells were treated with 10 and 20 µM of baicalin, baicalein, and wogonin for 48 h. n = 4 per group. ( B ) Gene expression of FGF21-regulating transcription factors in AML12 cells. Cells were treated with 10 and 20 µM of wogonin for 48 h. n = 4 per group. ( C ) Wogonin increased the protein levels of ATF4 in AML12 cells. Cells were treated with 10 and 20 µM of wogonin for 48 h. The protein bands were quantified. n = 4 per group. ( D ) Wogonin increased the expression of genes regulated by ATF4 in AML12 cells. Cells were treated with 10 and 20 µM of wogonin for 48 h. n = 4 per group. Data are represented as mean ± SD. * p < 0.05; ** p < 0.01. Comparisons among multiple groups were assessed using one-way ANOVA, followed by Tukey’s post hoc test.

Article Snippet: Wogonin (TOKYO CHEMICAL INDUSTRY CO., LTD, Tokyo, Japan, W0010), baicalin (Combi-Blocks, San Diego, CA, USA, QB-9653), and baicalein (BLDpharm, Shanghai, China, BD6296) were purchased.

Techniques: Expressing, Gene Expression