Journal: Neural Regeneration Research

Article Title: Blood–brain barrier disruption and neuroinflammation in the hippocampus of a cardiac arrest porcine model: Single-cell RNA sequencing analysis

doi: 10.4103/NRR.NRR-D-24-01269

Figure Lengend Snippet: Enhanced intercellular communication drives neuroinflammatory responses following cardiac arrest. (A) Heatmap illustrating the numbers of inferred interactions between different cell types based on LR pairs. (B) Chord diagram of the top 30 LR pairs between cell types. (C, D) Network diagrams of the changes in the number of intercellular communications between the Sham and ROSC6h groups (C) and the Sham and ROSC24h groups (D). Red represents an increase, and blue represents a decrease, in the number of interactions. (E) Visualization of enhanced LR pairs involved in microglia–neutrophil communication after cardiac arrest in the ROSC6h and ROSC24h groups vs. the Sham group. Each dot represents an LR pair, with its position indicating the group. Dot size corresponds to the P value (larger dots indicate lower P values), while the color represents the interaction probability, with the gradient ranging from purple (low probability) to yellow (high probability). (F) Secretion of resistin into the supernatant of the neutrophil–microglia co-culture system over time. (G) Comparison of resistin secretion, with and without OGD/R and the presence of neutrophils. (H) Immunofluorescence analysis and quantification of iNOS expression in microglia. The left panel shows representative images of iNOS (red), IBA1 (green, microglia marker), and DAPI (blue, nuclei), with merged images in the bottom row. The bar graph shows quantification of relative iNOS fluorescence intensity. Statistical significance determined by one-way analysis of variance followed by Tukey’s post hoc test; **** P < 0.0001. Scale bars: 20 μm. IBA1: Ionized calcium binding adaptor molecule 1; iNOS: inducible nitric oxide synthase; LR: ligand receptor; ns: not significant; OGD/R: oxygen-glucose deprivation/reperfusion; ROSC: return of spontaneous circulation.

Article Snippet: They were then incubated with the following primary antibodies at 4°C overnight: rabbit anti-ionized calcium binding adaptor molecule 1 (IBA1) (1:200 dilution; Abcam, Cambridge, UK, Cat# ab178846, RRID: AB_2636859); mouse anti-S100A8 (1:200; Proteintech, Wuhan, China, Cat# 66853-1-Ig, RRID: AB_2882193); rabbit anti-myelin basic protein (MBP) (1:50, Cell Signaling Technology, Danvers, MA, USA; Cat# 78896, RRID: AB_2799920); rabbit anti-myeloperoxidase (MPO) (1:100, Abcam, Cat# ab208670, RRID: AB_2864724); mouse anti-IBA1 (1:200, Santa Cruz Biotechnology, Santa Cruz, CA, USA, Cat# sc-32725, RRID: AB_667733); and rabbit anti-inducible nitric oxide synthase (iNOS) (1:200, Proteintech, Cat# 18985-1-AP, RRID: AB_2782960).

Techniques: Co-Culture Assay, Comparison, Immunofluorescence, Expressing, Marker, Fluorescence, Binding Assay

Journal: World Journal of Gastroenterology

Article Title: Cedrol ameliorates ulcerative colitis via myeloid differentiation factor 2-mediated inflammation suppression, with barrier restoration and microbiota modulation

doi: 10.3748/wjg.v32.i2.114057

Figure Lengend Snippet: Cedrol inhibited toll-like receptor 4-mediated mitogen-activated protein kinase and nuclear factor kappa B signaling in vivo and in vitro . A-D: Phosphorylated and total protein abundance of mitogen-activated protein kinase signaling molecules [extracellular regulated protein kinase (ERK)1/2, c-Jun N-terminal kinase (JNK), p38] in colonic tissues of mice ( n = 6); E: Cellular viability was quantified with the cell counting kit-8 method ( n = 3); F-H: Proinflammatory cytokine expression in RAW264.7 macrophages across experimental groups ( n = 3); I-K: Inducible nitric oxide synthase and cyclooxygenase-2 protein expression in RAW264.7 cells across groups ( n = 3); L-P: Phosphorylated and total protein expression of ERK1/2, JNK, p38 and nuclear factor kappa B p65 in RAW264.7 macrophages ( n = 3). Data are expressed as mean ± SD. a P < 0.05. b P < 0.01. c P < 0.001. d P < 0.0001. CE: Cedrol; DSS: Dextran sulfate sodium; ERK: Extracellular regulated protein kinase; p-ERK: Phospho-extracellular regulated protein kinase; JNK: C-Jun N-terminal kinase; p-JNK: Phospho-c-Jun N-terminal kinase; GAPDH: Glyceraldehyde-3-phosphate dehydrogenase; TNF: Tumor necrosis factor; IL: Interleukin; mRNA: Messenger RNA; LPS: Lipopolysaccharide; iNOS: Inducible nitric oxide synthase; COX-2: Cyclooxygenase-2.

Article Snippet: After blocking in 5% skim milk, membranes were incubated overnight at 4 °C with primary antibodies targeting: Phospho-extracellular regulated protein kinase (ERK) 1/2 (Thr202/Tyr204); ERK1/2; Phospho-c-Jun N-terminal kinase (JNK) (Thr183/Tyr185); JNK; Phospho-p38 (Thr180/Tyr182); P38, phospho-nuclear factor kappa B (NF-κB) p65 (Ser536); NF-κB p65 (Cell Signaling Technology, Daners, MA, United States); Cyclooxygenase-2 (COX-2); Inducible nitric oxide synthase (iNOS); Glyceraldehyde-3-phosphate dehydrogenase (Proteintech, Wuhan, Hubei Province, China).

Techniques: In Vivo, In Vitro, Quantitative Proteomics, Cell Counting, Expressing

Journal: World Journal of Gastroenterology

Article Title: Cedrol ameliorates ulcerative colitis via myeloid differentiation factor 2-mediated inflammation suppression, with barrier restoration and microbiota modulation

doi: 10.3748/wjg.v32.i2.114057

Figure Lengend Snippet: Cedrol treated dextran sulfate sodium-induced colitis by inhibiting inflammation, restoring the intestinal barrier, and rebalancing gut microbiota. TNF: Tumor necrosis factor; IL: Interleukin; DSS: Dextran sulfate sodium; LPS: Lipopolysaccharide; MD2: Myeloid differentiation factor 2; TLR4: Toll-like receptor 4; MAPK: Mitogen-activated protein kinase; ERK: Extracellular regulated protein kinase; JNK: C-Jun N-terminal kinase; NF-κB: Nuclear factor kappa B; iNOS: Inducible nitric oxide synthase; COX-2: Cyclooxygenase-2.

Article Snippet: After blocking in 5% skim milk, membranes were incubated overnight at 4 °C with primary antibodies targeting: Phospho-extracellular regulated protein kinase (ERK) 1/2 (Thr202/Tyr204); ERK1/2; Phospho-c-Jun N-terminal kinase (JNK) (Thr183/Tyr185); JNK; Phospho-p38 (Thr180/Tyr182); P38, phospho-nuclear factor kappa B (NF-κB) p65 (Ser536); NF-κB p65 (Cell Signaling Technology, Daners, MA, United States); Cyclooxygenase-2 (COX-2); Inducible nitric oxide synthase (iNOS); Glyceraldehyde-3-phosphate dehydrogenase (Proteintech, Wuhan, Hubei Province, China).

Techniques:

Journal: International Journal of Molecular Medicine

Article Title: Targeting of CIRP attenuates osteoarthritis progression via suppressing TLR4/NF-κB/NLRP3 signaling axis

doi: 10.3892/ijmm.2025.5674

Figure Lengend Snippet: CIRP promotes inflammatory response and ECM degradation in human chondrocytes. (A) Quantitative PCR analysis showing the effect of CIRP on the expression of inflammatory cytokines, including IL-6, IL-1β, TNF-α, iNOS and COX-2. (B) Western blotting showing the protein expression of iNOS and COX-2. (C and D) Quantitative PCR and western blotting showing the effect of CIRP on the expression of MMP1, MMP3, MMP13, ADAMTS5 and collagen Ⅱ. (E) Immunofluorescence analysis of collagen Ⅱ and MMP13 expression following CIRP treatment, with DAPI staining for nuclei (scale bar, 50 μ m). n=3, * P<0.05, ** P<0.01, *** P<0.001. CIRP, cold-inducible RNA-binding protein; ECM, extracellular matrix; iNOS, nitric oxide synthase; COX-2, cyclooxygenase-2; MMPs, matrix metalloproteinases; ADAMTS5, ADAM metallopeptidase with thrombospondin type 1 motif 5.

Article Snippet: TLR4 inhibitor TAK-242 was purchased from Shanghai Xianding Biotechnology Co., Ltd. Primary antibodies directed against inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), MMP1, MMP3, MMP13, Lamin B1, CD63, CD9, 130 kDa cis-Golgi matrix protein 1 (GM130) and GAPDH were purchased from Proteintech Group, Inc., ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) was purchased from ABclonal.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot, Immunofluorescence, Staining, RNA Binding Assay

Journal: International Journal of Molecular Medicine

Article Title: Targeting of CIRP attenuates osteoarthritis progression via suppressing TLR4/NF-κB/NLRP3 signaling axis

doi: 10.3892/ijmm.2025.5674

Figure Lengend Snippet: miR-145-5p elevation suppresses CIRP induced damage of chondrocytes. (A and B) Chondrocytes were transfected with miR-145-5p mimics or CIRP, IL-6, IL-1β, TNF-α, iNOS and COX-2 expression were analyzed by qPCR. (C) Chondrocytes were transfected with miR-145-5p mimics or CIRP, the mRNA expression of extracellular matrix proteins of chondrocytes were analyzed by qPCR. (D and E) Chondrocytes were transfected with miR-145-5p mimics or CIRP, the protein expression of iNOS, COX-2 and extracellular matrix proteins of chondrocytes were analyzed by western blotting. n=3, * P<0.05, ** P<0.01, *** P<0.001. miR, microRNA; CIRP, cold-inducible RNA-binding protein; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2; qPCR, quantitative PCR.

Article Snippet: TLR4 inhibitor TAK-242 was purchased from Shanghai Xianding Biotechnology Co., Ltd. Primary antibodies directed against inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), MMP1, MMP3, MMP13, Lamin B1, CD63, CD9, 130 kDa cis-Golgi matrix protein 1 (GM130) and GAPDH were purchased from Proteintech Group, Inc., ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) was purchased from ABclonal.

Techniques: Transfection, Expressing, Western Blot, RNA Binding Assay, Real-time Polymerase Chain Reaction

Journal: International Journal of Molecular Medicine

Article Title: Targeting of CIRP attenuates osteoarthritis progression via suppressing TLR4/NF-κB/NLRP3 signaling axis

doi: 10.3892/ijmm.2025.5674

Figure Lengend Snippet: Schematic illustration of targeting of CIRP attenuates osteoarthritis progression and the underlying mechanism. CIRP can be secreted in the form of exosomes and acts as a pro-inflammatory factor that activates the TLR4/NF-κB/NLRP3 signaling pathway, promoting the inflammatory response, ECM degradation and the progression of OA. Additionally, CIRP is identified as a target of miR-145, which inhibits its expression in OA. CIRP, cold-inducible RNA-binding protein; TLR4, Toll-like receptor 4; NLRP3, NLR family pyrin domain containing 3; OA, osteoarthritis; ECM, extracellular matrix; miR, microRNA; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2; MMPs, matrix metalloproteinases.

Article Snippet: TLR4 inhibitor TAK-242 was purchased from Shanghai Xianding Biotechnology Co., Ltd. Primary antibodies directed against inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), MMP1, MMP3, MMP13, Lamin B1, CD63, CD9, 130 kDa cis-Golgi matrix protein 1 (GM130) and GAPDH were purchased from Proteintech Group, Inc., ADAM metallopeptidase with thrombospondin type 1 motif 5 (ADAMTS5) was purchased from ABclonal.

Techniques: Expressing, RNA Binding Assay