ca1 tissues  (Proteintech)

Journal: Acta Neuropathologica Communications

Article Title: Reciprocal regulation of GPNMB/HIF-1α for Inhibition of neuronal ferroptosis in delayed encephalopathy after acute carbon monoxide poisoning

doi: 10.1186/s40478-025-02069-x

Figure Lengend Snippet: Fer-1 attenuates cognitive impairment of rats with DEACMP. ( A ) Escape latency was evaluated by MWM tests. ( B ) Swimming distance was evaluated by MWM tests. ( C ) Average swimming speed was assessed by MWM tests. ( D ) The number of platform crossing was recorded. For each day of training, data were averaged across five daily trials. ( E - F ) The histological changes in rat CA1 hippocampal tissues were assessed by H&E staining, and the morphology of hippocampal neurons was detected by Nissl staining. Scale bar, 25 μm. ( G ) The protein levels of SYP and PSD95 in rat CA1 hippocampal tissues were detected by western blot. N = 12 per group. ( A - D ) were analyzed by repeated measures two-way ANOVA followed by Bonferroni post hoc analysis ; ( F - G ) were analyzed by one-way ANOVA followed by Bonferroni’s multiple comparison tests . *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: The level of 8-OHdG in CA1 hippocampal tissues was measured using 8-OHdG ELISA kit (CSB-E10140h, Cusabio, Wuhan, China).

Techniques: Staining, Western Blot, Comparison

Journal: Acta Neuropathologica Communications

Article Title: Reciprocal regulation of GPNMB/HIF-1α for Inhibition of neuronal ferroptosis in delayed encephalopathy after acute carbon monoxide poisoning

doi: 10.1186/s40478-025-02069-x

Figure Lengend Snippet: Fer-1 attenuates inflammatory impairment and oxidative stress of rats with DEACMP. ( A ) The levels of pro-inflammatory cytokines in hippocampus CA1 area were measured by ELISA assay. ( B - C ) The levels of protein carbonyl content and 8-OHdG in hippocampus CA1 area were assessed using commercial kits. ( D - E ) The levels of GSH and MDA in hippocampus CA1 area were measured using commercial kits. ( F - I ) Iron accumulation in hippocampus CA1 area was detected by FerroOrange staining, and the immunoreactivity of GPX4 in hippocampus CA1 area was detected by IHC analysis. Red, FerroOrange; Blue, DAPI. Scale bar, 25 μm. N = 12 per group. ( A - E , G - I ) were analyzed by one-way ANOVA followed by Bonferroni’s multiple comparison tests . *, P < 0.05; **, P < 0.01; ***, P < 0.001

Article Snippet: The level of 8-OHdG in CA1 hippocampal tissues was measured using 8-OHdG ELISA kit (CSB-E10140h, Cusabio, Wuhan, China).

Techniques: Enzyme-linked Immunosorbent Assay, Staining, Comparison

Journal: Acta Neuropathologica Communications

Article Title: Reciprocal regulation of GPNMB/HIF-1α for Inhibition of neuronal ferroptosis in delayed encephalopathy after acute carbon monoxide poisoning

doi: 10.1186/s40478-025-02069-x

Figure Lengend Snippet: Expression pattern of GPNMB in rats with DECAMP. ( A ) The mRNA level of GPNMB in hippocampus CA1 area was detected by qRT-PCR. ( B ) The protein level of GPNMB in in hippocampus CA1 area was detected by western blot. N = 3. ( C ) Representative image of GPNMB staining in hippocampus CA1 area IF staining with quantitative analysis. Green, GPNMB; Blue, DAPI. Scale bar, 25 μm. N = 12 per group. ( A - C ) were analyzed by two tailed Student’s t -test. ***, P < 0.001

Article Snippet: The level of 8-OHdG in CA1 hippocampal tissues was measured using 8-OHdG ELISA kit (CSB-E10140h, Cusabio, Wuhan, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Staining, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Reciprocal regulation of GPNMB/HIF-1α for Inhibition of neuronal ferroptosis in delayed encephalopathy after acute carbon monoxide poisoning

doi: 10.1186/s40478-025-02069-x

Figure Lengend Snippet: STAT3 acts as a transcriptional activator of GPNMB. ( A ) The immunoreactivity of STAT3 in hippocampus CA1 area was assessed by IHC staining with quantitative analysis. Scale bar, 25 μm. ( B - C ) The protein levels of STAT3 and p-STAT3 in hippocampus CA1 area were detected by western blot. N = 3. ( D ) The mRNA level of STAT3 in PC-12 cells was detected by qRT-PCR. ( E ) Relative GPNMB promoter activity was measured by dual luciferase reporter assay. ( F ) The direct interaction between STAT3 and GPNMB promoter was detected by ChIP assay. Normal IgG served as a negative control. ( G - H ) The mRNA levels of STAT3 and GPNMB in PC-12 cells were detected by qRT-PCR. ( I ) The protein levels of STAT3 and GPNMB in PC-12 cells were detected by western blot. Experiments were performed in triplicate and error bars represent SD of a triplicate set of experiments. Data are shown as mean ± SD. ( A , C - I ) were analyzed by two tailed Student’s t -test. **, P < 0.01; ***, P < 0.001

Article Snippet: The level of 8-OHdG in CA1 hippocampal tissues was measured using 8-OHdG ELISA kit (CSB-E10140h, Cusabio, Wuhan, China).

Techniques: Immunohistochemistry, Western Blot, Quantitative RT-PCR, Activity Assay, Luciferase, Reporter Assay, Negative Control, Two Tailed Test

Journal: Acta Neuropathologica Communications

Article Title: Reciprocal regulation of GPNMB/HIF-1α for Inhibition of neuronal ferroptosis in delayed encephalopathy after acute carbon monoxide poisoning

doi: 10.1186/s40478-025-02069-x

Figure Lengend Snippet: Loss or gain of GPNMB takes actions on cognitive impairment, oxidative stress and neuronal ferroptosis of rats with DEACMP. ( A ) Escape latency was evaluated by MWM tests. ( B ) Swimming distance was evaluated by MWM tests. ( C ) Average swimming swspeed was assessed by MWM tests. ( D ) The number of platform crossing was recorded. For each day of training, data were averaged across five daily trials. ( E ) The morphology of hippocampal neurons was detected by Nissl staining. Scale bar, 25 μm. ( F ) Cell apoptosis in hippocampus CA1 area was assessed by TUNEL assay. Scale bar, 25 μm. ( G ) The levels of pro-inflammatory cytokines in hippocampus CA1 area were measured by ELISA assay. ( H - I ) The levels of protein carbonyl content and 8-OHdG in hippocampus CA1 area were assessed using commercial kits. ( J - K ) The levels of GSH and MDA in hippocampus CA1 area were measured using commercial kits. ( L ) The immunoreactivities of NeuN and GPX4 in hippocampus CA1 area were detected by IF staining. Green, NeuN; Red, GPX4; Blue, DAPI. Scale bar, 25 μm. ( M ) The protein levels of GPNMB and HIF-1α were detected by western blot. N = 12 per group. ( A - E , G - K , M ) were analyzed by one-way ANOVA followed by Bonferroni’s multiple comparison tests . *, P < 0.05; ***, P < 0.001

Article Snippet: The level of 8-OHdG in CA1 hippocampal tissues was measured using 8-OHdG ELISA kit (CSB-E10140h, Cusabio, Wuhan, China).

Techniques: Staining, TUNEL Assay, Enzyme-linked Immunosorbent Assay, Western Blot, Comparison

Journal: Molecular Neurobiology

Article Title: Huang Lian Jie Du Decoction Prevents Chronic Alcoholic Encephalopathy and Improves Gut Microbiota Imbalance in Mice

doi: 10.1007/s12035-025-05151-6

Figure Lengend Snippet: Histopathology of prefrontal cortex and hippocampus in mice. A H&E staining observation of prefrontal cortex (400 ×, scale bar, 50 μm, yellow arrows: dark-colored neurons). B H&E staining observation of hippocampal CA1 (400 ×, scale bar, 50 μm, yellow arrows: dark-colored neurons). C Toluidine blue staining observation of prefrontal cortex (400 ×, scale bar, 50 μm, red arrows: positive neurons). D Toluidine blue staining observation of hippocampal CA1 (400 ×, scale bar, 50 μm, red arrows: positive neurons). The data are expressed as the mean ± SD. Compared with the control group, ### P < 0.001, compared with the model group, *** P < 0.001, ** P < 0.01, * P < 0.05

Article Snippet: The hippocampal CA1 tissues were paraffin-embedded and sectioned, and the sections were immersed in citrate buffer and heated on high in a microwave oven for antigen repair with goat serum blocker, followed by the addition of primary antibody postsynaptic density protein 95 (PSD95) and synaptophysin (SYN) (Proteintech, Chicago, USA) overnight at 4 °C, secondary antibody (Servicebio, Wuhan, China) for 30 min at 37 °C, and a drop of DAPI for 10 min at room temperature.

Techniques: Histopathology, Staining, Control

Journal: Molecular Neurobiology

Article Title: Huang Lian Jie Du Decoction Prevents Chronic Alcoholic Encephalopathy and Improves Gut Microbiota Imbalance in Mice

doi: 10.1007/s12035-025-05151-6

Figure Lengend Snippet: Micropathologic changes of prefrontal cortex and hippocampal neurons in mice. A Transmission electron microscopic observations of prefrontal cortex (20,000 ×, Scale bar, 1 μm). B Transmission electron microscopic observations of prefrontal cortex hippocampal CA1 (20,000 ×, Scale bar, 1 μm). Red arrow, mitochondria; yellow arrow, rough endoplasmic reticulum; purple arrow, primary lysosome; blue arrow, secondary lysosomes

Article Snippet: The hippocampal CA1 tissues were paraffin-embedded and sectioned, and the sections were immersed in citrate buffer and heated on high in a microwave oven for antigen repair with goat serum blocker, followed by the addition of primary antibody postsynaptic density protein 95 (PSD95) and synaptophysin (SYN) (Proteintech, Chicago, USA) overnight at 4 °C, secondary antibody (Servicebio, Wuhan, China) for 30 min at 37 °C, and a drop of DAPI for 10 min at room temperature.

Techniques: Transmission Assay