Journal: Cell communication and signaling : CCS

Article Title: Free heme induces neuroinflammation and cognitive impairment by microglial activation via the TLR4/MyD88/NF-κB signaling pathway.

doi: 10.1186/s12964-023-01387-8

Figure Lengend Snippet: Fig. 3 Hemin induced microglial inflammation and neuronal apoptosis. ELISA analysis of TNF-α A, IL-1β B and IL-6 C in BV2 cells after hemin treatment for 24 h. D The gating strategy and representative FACS plots of apoptotic cells (Q2 + Q4) in the control group and Heme group of HT22 cells. E Quantification of the percentage of apoptotic cells among HT22 cells in the control group and Heme group of HT22 cells. F Representative immunofluorescence images of neurons (red) in the control group and Heme group of HT22 cells. Scale bar = 100 μm. Data are presented as means ± SEM. ***p < 0.001

Article Snippet: On the second day, the BV2 cell culture medium was replaced, and the cells were divided into 4 groups: 1) the Control group (BV2), in which BV2 cells were incubated with medium only for 24 h; 2) the Heme group (BV2 + Heme) in which BV2 cells were treated with 40 μM hemin (H9039, Sigma, USA) for 24 h; 3) the MyD88 inhibition group (BV2 + Heme + T6167923) in which BV2 cells were treated with 40 μM hemin and 2 μM MyD88 inhibitor (T6167923, #HY-19744, MedChemExpress, USA) for 24 h; and 4) the NF-κB inhibition group (BV2 + Heme + JSH-23), in which BV2 cells were treated with 40 μM hemin and 20 μM NF-κB inhibitor (JSH-23, #S7351, Selleck, USA) for 24 h. Then, the BV2 cells were harvested for subsequent experiments.

Techniques: Enzyme-linked Immunosorbent Assay, Control, Immunofluorescence

Journal: Cell communication and signaling : CCS

Article Title: Free heme induces neuroinflammation and cognitive impairment by microglial activation via the TLR4/MyD88/NF-κB signaling pathway.

doi: 10.1186/s12964-023-01387-8

Figure Lengend Snippet: Fig. 4 Microglia aggravated free heme-induced neuronal apoptosis. A BV2 cell and HT22 cell coculture system and four groups: 1) Neuron group (HT22) in which only HT22 cells were incubated for 24 h; 2) Neuron Heme group (HT22 + Heme) in which HT22 cells were treated with hemin for 24 h; 3) Neuron + microglia group (HT22 + BV2) in which HT22 cells cocultured with BV2 cells were incubated for 24 h; 4) Neuron + microglia Heme group (HT22 + BV2 + Heme) in which HT22 cells cocultured with BV2 cells were treated with hemin for 24 h. B Representative FACS plots of apoptotic cells (Q2 + Q4) in four groups. C Quantification of the percentage of apoptotic cells among HT22 cells in four groups. D Representative immunofluorescence images of nuclei (blue) and NeuN (red) staining among HT22 cells in four groups. Scale bar = 100 μm. E Quantitative analysis of NeuN + cells in four groups. Data are presented as means ± SEM. ***p < 0.001. *p < 0.05

Article Snippet: On the second day, the BV2 cell culture medium was replaced, and the cells were divided into 4 groups: 1) the Control group (BV2), in which BV2 cells were incubated with medium only for 24 h; 2) the Heme group (BV2 + Heme) in which BV2 cells were treated with 40 μM hemin (H9039, Sigma, USA) for 24 h; 3) the MyD88 inhibition group (BV2 + Heme + T6167923) in which BV2 cells were treated with 40 μM hemin and 2 μM MyD88 inhibitor (T6167923, #HY-19744, MedChemExpress, USA) for 24 h; and 4) the NF-κB inhibition group (BV2 + Heme + JSH-23), in which BV2 cells were treated with 40 μM hemin and 20 μM NF-κB inhibitor (JSH-23, #S7351, Selleck, USA) for 24 h. Then, the BV2 cells were harvested for subsequent experiments.

Techniques: Incubation, Immunofluorescence, Staining

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Histamine Pathway is a Target to Treat Hepatic Experimental Erythropoietic Protoporphyria

doi: 10.1016/j.jcmgh.2025.101463

Figure Lengend Snippet: Gene mRNA and protein expressions of heme biosynthesis enzymes. ( A ) ALAS1, ALAD, HMBS, PPOX, and FECH gene expressions are suppressed in the female fch/fch liver. ALAS1, UROD, and FECH gene expressions are suppressed in the male fch/fch liver. ( B ) Standard immunoblotting was carried with similar protein loading for each of the displayed panels. The ALAS1, FECH, ALAD, and UROD protein expression is suppressed in the fch/fch livers. Under the experimental conditions used in this study, HMW aggregates of ALAD and UROD (and not ALAS1 or FECH) were detected in the fch/fch livers. There is an upward shift in ALAD and UROD monomers as indicated by the arrowheads . Data are shown as mean ± SD (n = 2–3 livers per group). Statistical analysis was performed by 1-way ANOVA test, ∗∗∗ P < .001; ∗∗ P < .01; ∗ P < .05.

Article Snippet: Heme and porphyrin biosynthesis , ALAS-H , Santa Cruz Biotechnology , sc-137093 , Mouse.

Techniques: Western Blot, Expressing

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: The Histamine Pathway is a Target to Treat Hepatic Experimental Erythropoietic Protoporphyria

doi: 10.1016/j.jcmgh.2025.101463

Figure Lengend Snippet: Commercial Antibodies Used in the Study

Article Snippet: Heme and porphyrin biosynthesis , ALAS-H , Santa Cruz Biotechnology , sc-137093 , Mouse.

Techniques: