Journal: International Dental Journal

Article Title: 4632427E13Rik Facilitates Jaw Marrow-Derived Mesenchymal Stem Cells Osteogenesis and Angiogenesis Under Hypoxia Through miR-34a-5p/Aldoa/Hif-1α Pathway

doi: 10.1016/j.identj.2025.109364

Figure Lengend Snippet: Aldoa promotes osteogenesis and angiogenesis via the ERK/Hif-1α pathway. A, Alizarin red staining (14 days) and ALP staining (7 days). B, The gene expression levels of Runx2, Alp and Ocn were determined by qRT-PCR. β-actin was used as an internal reference gene. C, Tube formation assay was performed in the presence of CM. The scale bars represent 100 μm. D, The mRNA expression levels of Vegf and CD31 were analysed by quantitative RT-PCR. β-actin was used as an internal reference gene. E, Immunofluorescence staining was performed to detect Vegf expression after being treated by CM. The scale bars represent 25 μm. F, A Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis was performed to determine the top related pathways involving these differentially expressed mRNAs. G and H, The protein expression levels of Aldoa, Hif-1α and the phosphorylated of ERK, JUK and p38 among hypoxia, normoxia and hypoxia + si-Aldoa groups were determined by western blot. Semiquantitative analysis of the p-ERK/ERK, p-JUK/JUK and p38/p-p38 ratios were shown. β-actin was used as an internal reference gene. All data were expressed as means ± SD. * P < .05, ** P < .01.

Article Snippet: Rabbit anti-Aldoa (1:1000, 11217-1-AP), rabbit anti-Hif-1α (1:1000, ab179483), rabbit anti-Runx2 (1:1000, 8486S), rabbit anti-CD31 (1:1000, 77699S), rabbit anti-Ocn (1:1000, bs-4917R), rabbit anti-Alp (1:1000, bs-1535R), rabbit anti-Vegf (1:1000, bs-1313R), rabbit anti-ERK (1:1000, bsm-33337M), rabbit anti-p-ERK (1:1000, bs-1646R), rabbit anti-p38 (1:1000, bs-0637R), rabbit anti-p-p38 (1:1000, bs-0636R), rabbit anti-JNK (1:1000, bs-2592R), rabbit anti-p-JNK (1:1000, bs-1640R), and mouse anti-β-actin (1:2000, AF7018) were purchased from Proteintech, Abcam, Cell Signalling Technology, and Bioss, respectively.

Techniques: Staining, Gene Expression, Quantitative RT-PCR, Tube Formation Assay, Expressing, Immunofluorescence, Western Blot

Journal: International Dental Journal

Article Title: 4632427E13Rik Facilitates Jaw Marrow-Derived Mesenchymal Stem Cells Osteogenesis and Angiogenesis Under Hypoxia Through miR-34a-5p/Aldoa/Hif-1α Pathway

doi: 10.1016/j.identj.2025.109364

Figure Lengend Snippet: Aldoa promotes osteogenesis and angiogenesis via the ERK/Hif-1α pathway. A, Alizarin red staining (14 days) and ALP staining (7 days). B, The gene expression levels of Runx2, Alp and Ocn were determined by qRT-PCR. β-actin was used as an internal reference gene. C, Tube formation assay was performed in the presence of CM. The scale bars represent 100 μm. D, The mRNA expression levels of Vegf and CD31 were analysed by quantitative RT-PCR. β-actin was used as an internal reference gene. E, Immunofluorescence staining was performed to detect Vegf expression after being treated by CM. The scale bars represent 25 μm. F, A Kyoto Encyclopaedia of Genes and Genomes (KEGG) enrichment analysis was performed to determine the top related pathways involving these differentially expressed mRNAs. G and H, The protein expression levels of Aldoa, Hif-1α and the phosphorylated of ERK, JUK and p38 among hypoxia, normoxia and hypoxia + si-Aldoa groups were determined by western blot. Semiquantitative analysis of the p-ERK/ERK, p-JUK/JUK and p38/p-p38 ratios were shown. β-actin was used as an internal reference gene. All data were expressed as means ± SD. * P < .05, ** P < .01.

Article Snippet: Rabbit anti-Aldoa (1:1000, 11217-1-AP), rabbit anti-Hif-1α (1:1000, ab179483), rabbit anti-Runx2 (1:1000, 8486S), rabbit anti-CD31 (1:1000, 77699S), rabbit anti-Ocn (1:1000, bs-4917R), rabbit anti-Alp (1:1000, bs-1535R), rabbit anti-Vegf (1:1000, bs-1313R), rabbit anti-ERK (1:1000, bsm-33337M), rabbit anti-p-ERK (1:1000, bs-1646R), rabbit anti-p38 (1:1000, bs-0637R), rabbit anti-p-p38 (1:1000, bs-0636R), rabbit anti-JNK (1:1000, bs-2592R), rabbit anti-p-JNK (1:1000, bs-1640R), and mouse anti-β-actin (1:2000, AF7018) were purchased from Proteintech, Abcam, Cell Signalling Technology, and Bioss, respectively.

Techniques: Staining, Gene Expression, Quantitative RT-PCR, Tube Formation Assay, Expressing, Immunofluorescence, Western Blot

Journal: Journal of Neuroinflammation

Article Title: Esketamine alleviates COPD-depression comorbidity in rats via MAPK/NF-κB inhibition and gut-lung-brain axis modulation

doi: 10.1186/s12974-026-03699-1

Figure Lengend Snippet: Activation of MAPK/NF-κB signaling accompanied by systemic inflammation and oxidative stress in the COPD-depression comorbid rat model. a , Serum cytokines and oxidative stress markers, including IL-6, IL-8, TNF-α, IL-10, MPO, SOD, and MDA ( n = 7). b , Representative western blots of lung tissue showing p-p38, p38, p-JNK, JNK, p-p65, p65, and GAPDH, with quantification of phosphorylation ratios ( n = 3). c , Representative western blots of hippocampal tissue showing p-p38, p38, p-JNK, JNK, p-p65, p65, and GAPDH, with quantification of phosphorylation ratios ( n = 3). d , Representative immunohistochemical staining of lung sections for p-p38, p-JNK, p-p65, with quantitative analysis of mean optical density ( n = 3). e , Representative immunohistochemical staining of hippocampal sections for p-p38, p-JNK, and p-p65, with quantitative analysis of mean optical density ( n = 3). Con, control; COPD, COPD model; YY, depression model; CY, comorbid model. Data are shown as mean ± SD. a : one-way ANOVA with Bonferroni post-hoc correction was used (* P < 0.05, ** P < 0.01 vs. Con); Welch test with Dunnett’s T3 correction was used where appropriate ( △ P < 0.05 vs. Con). b - e : one-way ANOVA with LSD post-hoc correction (* P < 0.05 vs. Con)

Article Snippet: Sections were incubated overnight at 4 °C with phospho-p38 (Bioss, bs-0636R, 1:200 for lung / 1:100 for hippocampus), phospho-JNK (Proteintech, 80024-1-RR, 1:200), or phospho-p65 (Affinity, AF2006, 1:200 for lung / 1:150 for hippocampus) at empirically optimized dilutions for lung and brain, followed by HRP-conjugated secondary antibodies (30 min, 37 °C).

Techniques: Activation Assay, Western Blot, Phospho-proteomics, Immunohistochemical staining, Staining, Control

Journal: Journal of Neuroinflammation

Article Title: Esketamine alleviates COPD-depression comorbidity in rats via MAPK/NF-κB inhibition and gut-lung-brain axis modulation

doi: 10.1186/s12974-026-03699-1

Figure Lengend Snippet: Esketamine suppresses MAPK/NF-κB signaling and restores inflammatory and oxidative balance in comorbid COPD-depression rats. a , Serum cytokines and oxidative stress markers, including IL - 6, IL - 8, TNF - α, IL - 10, MDA, MPO, and SOD ( n = 7). b , Representative western blots of lung tissue showing p-p38, p38, p - JNK, JNK, p - p65, p65, and GAPDH, with quantification of phosphorylation ratios ( n = 3). c , Representative western blots of hippocampal tissue showing p-p38, p38, p-JNK, JNK, p-p65, p65, and GAPDH, with quantification of phosphorylation ratios ( n = 3). d , Representative lung sections stained for p-p38, p - JNK, and p-p65, with quantification of mean optical density (AOD) ( n = 3). e , Representative hippocampal sections stained for p - p38, p - JNK, and p-p65, with quantification of mean optical density (AOD) ( n = 3).Con, control; CY, comorbid model; CYK, CY rats treated with esketamine; CYKD, CY rats treated with esketamine and Diprovocim. Data are expressed as mean ± SD. a : one-way ANOVA with Bonferroni post-hoc correction was used (** P < 0.01 vs. Con; ## P < 0.01 vs. CY; ▲ P < 0.05, ▲ ▲ P < 0.01 vs. CYK). b - e : one-way ANOVA with LSD post-hoc correction was used (* P < 0.05 vs. Con; # P < 0.05 vs. CY; ▲ P < 0.05 vs. CYK)

Article Snippet: Sections were incubated overnight at 4 °C with phospho-p38 (Bioss, bs-0636R, 1:200 for lung / 1:100 for hippocampus), phospho-JNK (Proteintech, 80024-1-RR, 1:200), or phospho-p65 (Affinity, AF2006, 1:200 for lung / 1:150 for hippocampus) at empirically optimized dilutions for lung and brain, followed by HRP-conjugated secondary antibodies (30 min, 37 °C).

Techniques: Western Blot, Phospho-proteomics, Staining, Control

Journal: Journal of Neuroinflammation

Article Title: Esketamine alleviates COPD-depression comorbidity in rats via MAPK/NF-κB inhibition and gut-lung-brain axis modulation

doi: 10.1186/s12974-026-03699-1

Figure Lengend Snippet: Cellular validation of MAPK/NF-κB signaling in alveolar macrophages and microglia. a - b , Dose-dependent effects of CSE and esketamine on NR8383 cell viability (CCK-8 assay). c , Western blot analysis of p-p38, p38, p-JNK, JNK, p-p65, p65, and GAPDH in NR8383 cells, with quantification of phosphorylation ratios ( n = 3). d - f , Grouped NR8383 assays showing cell viability (CCK-8) and release of inflammatory cytokines (IL-6, IL-8, TNF-α, IL-10) and oxidative stress markers (MDA, MPO, SOD) ( n = 3). g - h , Dose-dependent effects of LPS + CSE and esketamine on HAPI cell viability (CCK-8 assay). i , Western blot analysis of p-p38, p38, p-JNK, JNK, p-p65, p65, and GAPDH in HAPI cells, with quantification of phosphorylation ratios ( n = 3). j - l , Grouped HAPI assays showing cell viability (CCK-8) and release of inflammatory cytokines and oxidative stress markers ( n = 3). Con, control; CSE, cigarette smoke extract; LPS, lipopolysaccharide. Data are expressed as mean ± SD; * P < 0.05 vs. Con; # P < 0.05 vs. CSE or LPS + CSE; ▲ P < 0.05 vs. CSE + Esketamine or LPS + CSE + Esketamine (LSD correction)

Article Snippet: Sections were incubated overnight at 4 °C with phospho-p38 (Bioss, bs-0636R, 1:200 for lung / 1:100 for hippocampus), phospho-JNK (Proteintech, 80024-1-RR, 1:200), or phospho-p65 (Affinity, AF2006, 1:200 for lung / 1:150 for hippocampus) at empirically optimized dilutions for lung and brain, followed by HRP-conjugated secondary antibodies (30 min, 37 °C).

Techniques: Biomarker Discovery, CCK-8 Assay, Western Blot, Phospho-proteomics, Control