inhibition activity against k pneumoniae atcc 13883  (ATCC)

Journal: Frontiers in Immunology

Article Title: Repetitive transcranial magnetic stimulation alleviates neuropathic pain via microglial polarization by modulating the METTL3/NMDAR2B/NLRP3 pathway

doi: 10.3389/fimmu.2025.1666920

Figure Lengend Snippet: In vivo (A–E) and in vitro (F–J) effects of rTMS on METTL3, m6A methylation, NMDAR2B, and YTHDF1. (A) METTL3, NMDAR2B, YTHDF1 protein bands in the DLPFC. (B–D) Western blot analysis of METTL3, YTHDF1, and NMDAR2B protein expression in the DLPFC. (E) ELISA analysis of the methylation modification level of m6A in the DLPFC. The number of samples in each group is 3. (F) METTL3, NMDAR2B, YTHDF1 protein bands in BV2 cells. (G–J) Western blot analysis of METTL3, YTHDF1, and NMDAR2B protein expression levels in BV2 cells. (P) ELISA analysis of the methylation modification level of m6A in BV2 cells. The number of samples in each group was 3. The results was analyzed with one-way ANOVA followed by Tukey’s post-hoc test, *p < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: After seeding, BV2 cells were cultured in complete DMEM for 24 h. Subsequently, specific pharmacological activation and inhibition targeting METTL3 (METTL3 activator-1; HY-W037893; MedChemExpress; STM2457; T9060; Targetmol) and NMDAR2B ( d -Serine; HY-100808; MedChemExpress; D-AP5; HY-100714A; MedChemExpress) were added to the culture medium, and their effects were verified.

Techniques: In Vivo, In Vitro, Methylation, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Modification

Journal: Frontiers in Immunology

Article Title: Repetitive transcranial magnetic stimulation alleviates neuropathic pain via microglial polarization by modulating the METTL3/NMDAR2B/NLRP3 pathway

doi: 10.3389/fimmu.2025.1666920

Figure Lengend Snippet: The roles of METTL3 and NMDAR2B in the anti-inflammatory effects of magnetic stimulation in vitro . (A) Effects of inhibition and overexpression of METTL3 on the protein bands of METTL3, NMDAR2B, and YTHDF1. (B–D) Western blot analyses of METTL3, YTHDF1, and NMDAR2B protein expression. (E) Effects of inhibition and overexpression of NMDAR2B on the protein bands of METTL3, NMDAR2B, and YTHDF1 (F–H) Western blot analyses of METTL3, YTHDF1, and NMDAR2B protein expression. (I–R) ELISA was used to detect the methylation modification level of m6A, NLRP3, IL-6, TNF-α, and IL-10. The number of samples in each group was 3. The results was analyzed with one-way ANOVA followed by Tukey’s post-hoc test, *p < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: After seeding, BV2 cells were cultured in complete DMEM for 24 h. Subsequently, specific pharmacological activation and inhibition targeting METTL3 (METTL3 activator-1; HY-W037893; MedChemExpress; STM2457; T9060; Targetmol) and NMDAR2B ( d -Serine; HY-100808; MedChemExpress; D-AP5; HY-100714A; MedChemExpress) were added to the culture medium, and their effects were verified.

Techniques: In Vitro, Inhibition, Over Expression, Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Methylation, Modification

Journal: Frontiers in Immunology

Article Title: Repetitive transcranial magnetic stimulation alleviates neuropathic pain via microglial polarization by modulating the METTL3/NMDAR2B/NLRP3 pathway

doi: 10.3389/fimmu.2025.1666920

Figure Lengend Snippet: The roles of METTL3 and NMDAR2B in the regulation of microglial cell polarization by magnetic stimulation in vitro . (A–C, G–I) Immunofluorescence staining of Iba-1, CD86 and CD206. (D–F, J–L) Proportion of positive cells for target proteins. The number of samples in each group was 3. The results was analyzed with one-way ANOVA followed by Tukey’s post-hoc test, *p < 0.05, **P < 0.01, ***P < 0.001 and ****P < 0.0001.

Article Snippet: After seeding, BV2 cells were cultured in complete DMEM for 24 h. Subsequently, specific pharmacological activation and inhibition targeting METTL3 (METTL3 activator-1; HY-W037893; MedChemExpress; STM2457; T9060; Targetmol) and NMDAR2B ( d -Serine; HY-100808; MedChemExpress; D-AP5; HY-100714A; MedChemExpress) were added to the culture medium, and their effects were verified.

Techniques: In Vitro, Immunofluorescence, Staining

Journal: American Journal of Cancer Research

Article Title: Macrophage-derived exosomes induce M2 microglial polarization to alleviate bone cancer pain

doi: 10.62347/BCLF6941

Figure Lengend Snippet: Exosomes suppressed TLR4/NF-κB signaling pathway related protein expression in BV2 microglial cells. A. Western blot analysis of TLR4, p-p65, p65, p-IκBα, and IκBα protein expression in BV2 microglial cells. LPS treatment significantly increased the expression of TLR4, p-p65, and p-IκBα, indicating activation of the TLR4/NF-κB pathway. Exosome treatment in the LPS + Exosome group reduced the expression of these proteins. β-actin was used as an internal control. B. ELISA analysis of inflammatory cytokines (IL-6, TNF-α, MCP-1) in the culture supernatants of BV2 cells. LPS treatment significantly increased the levels of these cytokines, while exosome treatment reduced their expression. Note: Toll-like receptor 4 (TLR4), Phosphorylated p65 (p-p65), p65 subunit of nuclear factor kappa-light-chain-enhancer of activated B cells (p65), Phosphorylated inhibitor of κB alpha (p-IκBα), Inhibitor of κB alpha (IκBα), Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB), Enzyme-linked immunosorbent assay (ELISA). Data are presented as mean ± SD. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05 vs. Control or LPS group (n=3).

Article Snippet: However, in the LPS + Exosome group, the expression of these proteins was significantly reduced, suggesting that exosomes inhibited LPS-induced TLR4/NF-κB pathway activation.

Techniques: Expressing, Western Blot, Activation Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: American Journal of Cancer Research

Article Title: Macrophage-derived exosomes induce M2 microglial polarization to alleviate bone cancer pain

doi: 10.62347/BCLF6941

Figure Lengend Snippet: Exosomes regulated M1/M2 polarization and the TLR4/NF-κB signaling pathway in the spinal cord of rats with bone cancer pain. A. Immunofluorescence staining showing the localization and internalization of red PKH67-labeled exosomes (red) in microglia and macrophages in the gray matter region of the spinal cord. Nuclei were stained with DAPI (blue), and merged images confirm exosome uptake in the BCP + Exo group (× 600). B. Western blot analysis of M1 polarization markers (iNOS, CD86, CD68) in the Sham, BCP , and BCP + Exo groups. M1 markers were significantly upregulated in the BCP group, while exosome treatment reduced their expression. C. Western blot analysis of M2 polarization markers (CD206, Arg-1). M2 markers were downregulated in the BCP group, and exosome treatment significantly upregulated their expression. D. Western blot analysis of TLR4, p-p65, and p-IκBα. The TLR4/NF-κB pathway was activated in the BCP group, and exosome treatment inhibited the activation of this pathway. Note: Inducible nitric oxide synthase (iNOS), Cluster of differentiation 86 (CD86), Cluster of differentiation 206 (CD206), Arginase 1 (Arg-1), Phosphorylated p65 (p-p65), Phosphorylated inhibitor of κB alpha (p-IκBα), Lipopolysaccharide (LPS), Western blot (WB). Data are presented as mean ± SD. ****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05 vs. Sham or BCP group (n=3).

Article Snippet: However, in the LPS + Exosome group, the expression of these proteins was significantly reduced, suggesting that exosomes inhibited LPS-induced TLR4/NF-κB pathway activation.

Techniques: Immunofluorescence, Staining, Labeling, Western Blot, Expressing, Activation Assay