Journal: International Journal of Molecular Medicine

Article Title: Propofol upregulates MFG-E8 in BV2 cells to inhibit pyroptosis mediated by the NF-κB/NLRP3 pathway, thereby ameliorating ischemic-reperfusion neuronal injury

doi: 10.3892/ijmm.2026.5786

Figure Lengend Snippet: Overexpression of MFG-E8 suppresses NF-κB and NLRP3 activation in OGD/R-induced BV2 cells. (A) Western blotting protein bands for MFG-E8, p-NF-κB, NF-κB, and GAPDH (internal reference). (B and C) Following OGD/R, western blotting revealed decreased MFG-E8 expression (B) and elevated p-NF-κB/NF-κB levels (C) in BV2 cells. (D) Western blotting protein bands for MFG-E8 and GAPDH (internal reference). (E) Following transfection of OE-MFG-E8 into BV2 cells, MFG-E8 expression was elevated markedly. (F) Western blotting protein bands for NLRP3, ASC, p-NF-κB, NF-κB, and GAPDH (internal reference). (G and H) Following transfection with OE-MFG-E8, p-NF-κB/NF-κB levels (G), and NLRP3 and ASC levels (H) were detected using western blotting. (I) Western blotting protein bands for NLRP3, ASC, p-NF-κB, NF-κB, and GAPDH (internal reference). Following treatment with the NF-κB activator LPS, p-NF-κB/NF-κB (J), and NLRP3 and ASC levels (K) were detected by western blotting. (L) Western blotting protein bands for p-NF-κB, NF-κB, and GAPDH (internal reference). (M) Following transfection with OE-MFG-E8, p-NF-κB/NF-κB expression was decreased, whereas treatment with LPS increased p-NF-κB/NF-κB levels. (N) Immunofluorescence staining of NLRP3 in BV2 cells (magnification, ×40; scale bar, 50 μ m). (O) Immunofluorescence staining of ASC in BV2 cells (magnification, ×40; scale bar, 50 μ m). (P) Immunofluorescence revealed OE-MFG-E8 reduced the fluorescence intensity of NLRP3 and ASC, while LPS diminished the effect of MFG-E8 overexpression. (Q) Western blotting protein bands for NLRP3, ASC, cleaved-caspase-1, pro-caspase-1, GSDMD-N, GSDMD, IL-1β, IL-18, and GAPDH (internal reference). (R-T) Western blotting revealed that OE-MFG-E8 reduced cleaved-caspase-1/pro-caspase-1 levels (R), GSDMD-N/GSDMD levels (S), NLRP3, ASC, IL-1β, and IL-18 levels (T) in OGD/R-induced BV2 cells, LPS attenuated the impact of OE-MFG-E8. ** P<0.01. PPF, Propofol; OGD/R, Oxygen-glucose deprivation/reoxygenation; ASC, apoptosis-associated speck-like protein containing a CARD; MFG-E8, milk fat globule-EGF factor 8; GSDM, gasdermin; OE, overexpression; p-, phosphorylated; LPS, lipopolysaccharide; NC, negative control; ns, not significant.

Article Snippet: For the OGD/R + OE-MFG-E8 + LPS (NF-κB agonist) group, cells were transfected with OE-MFG-E8, treated with OGD and exposed to LPS (1 μ g/ml; cat. no. HY-D1056, MedChemExpress) during reoxygenation ( ).

Techniques: Over Expression, Activation Assay, Western Blot, Expressing, Transfection, Immunofluorescence, Staining, Fluorescence, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: Propofol upregulates MFG-E8 in BV2 cells to inhibit pyroptosis mediated by the NF-κB/NLRP3 pathway, thereby ameliorating ischemic-reperfusion neuronal injury

doi: 10.3892/ijmm.2026.5786

Figure Lengend Snippet: PPF suppresses pyroptosis caused by NF-κB/NLRP3 signaling by upregulating MFG-E8. (A) Western blotting protein bands for MFG-E8, p-NF-κB, NF-κB, and GAPDH (internal reference) in BV2 cells. (B) Western blotting revealed that PPF increased MFG-E8 and decreased p-NF-κB/NF-κB levels (C) in BV2 cells. (D) Western blotting for MFG-E8 and GAPDH in BV2 cells. (E) Following transfection of si-MFG-E8 into BV2 cells, MFG-E8 protein levels were significantly decreased. (F) Dual luciferase reporter gene assay confirmed that NF-κB is the target of MFG-E8. (G) Western blotting protein bands for p-NF-κB, NF-κB, and GAPDH (internal reference) in BV2 cells. (H) si-MFG-E8 attenuated the effects of PPF, resulting in elevated p-NF-κB/NF-κB levels. (I) Yo-Pro-1 and Hoechst 33342 staining for assessing the pyroptosis levels in BV-2 cells (magnification, ×40; scale bar, 50 μ m). (J) Yo-Pro-1 and Hoechst 33342 staining showed that PPF decreased Yo-Pro-1 positivity, while silencing MFG-E8 increased Yo-Pro-1 positivity. ELISA showed that PPF decreased TNF-α (K) and IL-1β (L) levels, silencing MFG-E8 reversed this effect. (M and N) ELISA showed that PPF raised IL-10 levels (M) and decreased IL-6 levels (N), silencing MFG-E8 reversed this effect. (O) Western blotting protein bands for NLRP3, ASC, cleaved-caspase-1, pro-caspase-1, GSDMD-N, GSDMD, IL-1β, IL-18, and GAPDH (internal reference) in BV2 cells. (P-R) Western blot analysis indicated that PPF decreased NLRP3, ASC, IL-1β, and IL-18 levels (P), GSDMD-N/GSDMD levels (Q), cleaved-caspase-1/pro-caspase-1 levels (R), silencing MFG-E8 increased these protein levels. ** P<0.01. PPF, propofol; MFG-E8, milk fat globule-EGF factor 8; OGD/R, Oxygen-glucose deprivation/reoxygenation; ASC, apoptosis-associated speck-like protein containing a CARD; GSDM, gasdermin; OE, overexpression; p-, phosphorylated; LPS, lipopolysaccharide; NC, negative control; ns, not significant; si, small interfering; WT, wild-type; MUT, mutant.

Article Snippet: For the OGD/R + OE-MFG-E8 + LPS (NF-κB agonist) group, cells were transfected with OE-MFG-E8, treated with OGD and exposed to LPS (1 μ g/ml; cat. no. HY-D1056, MedChemExpress) during reoxygenation ( ).

Techniques: Western Blot, Transfection, Luciferase, Reporter Gene Assay, Staining, Enzyme-linked Immunosorbent Assay, Over Expression, Negative Control, Mutagenesis

Journal: International Dental Journal

Article Title: Pulpitis Transiently Affect Hepatic Bone Morphogenetic Protein 9 Expression by Lipopolysaccharide

doi: 10.1016/j.identj.2026.109435

Figure Lengend Snippet: UNC0642 restored BMP9 expression in C57 mice and binding of H3K9me2 to the promoter region of BMP9 gene. (A) The expression of BMP9 in the DMSO-PO group was decreased, whereas that in the UNC0642-PO group was upregulated and similar to that in the DMSO control group (WB). (B) The RT-qPCR results are consistent with the IHC results. (C and D) The WB results were consistent with the IHC results ( n = 6). The groups 1 to 3 correspond to the DMSO group, DMSO-PO group, and UNC0642-PO group, respectively. (E) After stimulation of mHSCs with P.g LPS, the enrichment rate of H3K9me2 in the promoter region of BMP9 gene was significantly increased (ChIP) ( n = 3). (F) Full-text pattern diagram. LPS from pulpitis reached the liver through the bloodstream, stimulating hepatic stellate cells and regulating the expression of BMP9 in the liver via H3K9 dimethylation. DMSO represents the working fluid of inhibitors in vivo: DMSO + 40% PEG300 + 5% Tween80 + double-distilled H2O; LPS, samples stimulated with P.g LPS; me2, dimethylation; NS, normal sample; PO, pulp opening. Data are presented as mean ± SD; * P < .05. Black scale: 100 μm, white scale: 50 μm.

Article Snippet: For in vitro experiments, mHSCs were cultured into 6-cm dishes or 24-well plates and divided into a dimethyl sulfoxide (DMSO) control group; DMSO + LPS group; and 0.5, 1, and 5 μM UNC0642 (Selleck) groups.

Techniques: Expressing, Binding Assay, Control, Quantitative RT-PCR, In Vivo