Journal: Scientific Reports

Article Title: Cold inducible RNA binding protein promotes fibroblast activation and its inhibition represents a potential therapeutic target in pulmonary fibrosis

doi: 10.1038/s41598-026-39649-3

Figure Lengend Snippet: CIRBP promotes collagen production and proliferation via TLR2/4-dependent IL-6 signaling. ( A ) IL-6 concentrations in the culture supernatants of primary lung fibroblasts treated with rmCIRBP, with or without C23, TLR4-I, or TLR2-I, measured by ELISA. ( B ) Soluble collagen levels in the supernatants following treatment with rmCIRBP ± anti-IL-6 antibody. ( C ) Proliferative activity of fibroblasts assessed by the WST assay after treatment with rmCIRBP and/or anti-IL-6 antibody. ( D ) Soluble collagen production in fibroblasts treated with rmCIRBP and either TLR4-I or TLR2-I. ( E ) Fibroblast proliferation measured using the WST assay following cotreatment with rmCIRBP and TLR4-I or TLR2-I. ( F ) Schematic representation of the proposed mechanism: CIRBP, which is highly expressed in fibrotic lungs, stimulates IL-6 production via TLR2/4, promoting collagen secretion and fibroblast proliferation in an autocrine manner and contributing to the progression of tissue fibrosis. Data are presented as mean ± SEM. *P < 0.05 using one-way ANOVA and Tukey’s multiple comparison test.

Article Snippet: Following starvation, the cells were treated with vehicle control, rmCIRBP (1.0 μg/mL), C23 (0.1 μg/mL), TLR4 inhibitor (2.76 μM; CLI-095, tlrl-cli95-4, InvivoGen, San Diego, CA, USA), TLR2 inhibitor (100 μM; TL2-C29, inh-c29, InvivoGen), or combinations thereof.

Techniques: Enzyme-linked Immunosorbent Assay, Activity Assay, WST Assay, Comparison

Journal: Scientific Reports

Article Title: RBP4 interferes with tongue squamous cell carcinoma progression by inhibiting the PI3K/AKT signaling pathway and promoting macrophage M1-type polarization

doi: 10.1038/s41598-026-39915-4

Figure Lengend Snippet: RBP4 promotes macrophage M1 polarization by activating the TLR4/NF-κB pathway. ( a ): KEGG Bar Charts of human recombinant RBP4 protein-treated M0 macrophage cells after transcriptome sequencing. ( b ): Correlation analysis of RBP4 and TLR4 protein expression using Spearman and Pearson methods. ( c ): Western blotting assay to detect the effect of human recombinant RBP4 on macrophage TLR4/NF-KB pathway and the change of the effect of RBP4 action after the addition of TAK242. ( d , e ): qRT-PCR and western blotting are used to detect the effect of the human recombinant RBP4 protein on the macrophage phenotype and the effect of RBP4 action after the addition of TAK242. ( f , g ): Western blotting experiments are used to detect the effects of changes in RBP4 expression levels on the macrophage TLR4/NF-κB pathway in a Transwell co-culture system. ( h ): Western blotting experiments are used to detect the effects of tumor-secreted RBP4 on the macrophage TLR4/NF-κB pathway after the addition of TAK242. Changes in activation level. ( i ): Western blotting experiments are used to detect the effect of tumor-secreted RBP4 on macrophage phenotype after addition of TAK242 in Transwell co-culture system. TAK242 treatment blocks RBP4 overexpression induced by TSCC cells in the macrophage TLR4/p-NF-κB upregulation and M1-type polarization. KEGG, Kyoto Encyclopedia of Genes and Genomes; TSCC, tongue squamous cell carcinoma. (*Compared with Control, * p < 0.05, ** p < 0.01, *** p < 0.001,*** p < 0.0001).

Article Snippet: Transcriptome sequencing showed that RBP4 activated the TLRs/NF-κB pathway (Fig. a) and that TLR4 and RBP4 expression were positively correlated (Spearman, Fig. b).Western blotting showed elevated expression of TLR4 and p-NF-κB in macrophages incorporating the human recombinant RBP4 protein (with no NF-κB changes) (Fig. c).QRT-PCR and Western blotting confirmed that RBP4 significantly up-regulated M1 markers (CD86, iNOS) and inhibited M2 markers (CD206, Arg-1) (Fig. d, e), whereas the TLR4 inhibitor TAK242 (0.2 Mm, HY-11109, MCE) reversed this effect ( Fig. c, e).

Techniques: Recombinant, Sequencing, Expressing, Western Blot, Quantitative RT-PCR, Co-Culture Assay, Activation Assay, Over Expression, Control

Journal: Scientific Reports

Article Title: RBP4 interferes with tongue squamous cell carcinoma progression by inhibiting the PI3K/AKT signaling pathway and promoting macrophage M1-type polarization

doi: 10.1038/s41598-026-39915-4

Figure Lengend Snippet: Effect of RBP4 on macrophage phenotype. ( a ): Animals were grouped according to the injection of different cells, and the tumor volume, growth curve, and mass were measured and recorded. ( b ): The levels of CD86 and CD206 cell infiltration in the sections of transplanted tumors from each group are analyzed using immunofluorescence staining. ( c ): Changes in the activation level of the TLR4/NF-KB pathway in each group of transplanted tumor tissues are detected using Western blotting. ( d ): Mechanistic diagram of the effects of changes in RBP4 levels on the proliferation, migration, and invasion abilities of TSCC cells and the effects of tumor-secreted RBP4 on the phenotype of TAMs. TAMs, tumor-associated macrophages; TSCC, tongue squamous cell carcinoma. (*Compared with RAW264.7 + Control, * p < 0.05, ** p < 0.01).

Article Snippet: Transcriptome sequencing showed that RBP4 activated the TLRs/NF-κB pathway (Fig. a) and that TLR4 and RBP4 expression were positively correlated (Spearman, Fig. b).Western blotting showed elevated expression of TLR4 and p-NF-κB in macrophages incorporating the human recombinant RBP4 protein (with no NF-κB changes) (Fig. c).QRT-PCR and Western blotting confirmed that RBP4 significantly up-regulated M1 markers (CD86, iNOS) and inhibited M2 markers (CD206, Arg-1) (Fig. d, e), whereas the TLR4 inhibitor TAK242 (0.2 Mm, HY-11109, MCE) reversed this effect ( Fig. c, e).

Techniques: Injection, Immunofluorescence, Staining, Activation Assay, Western Blot, Migration, Control

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a Volcano plot comparing DEGs for RNA-seq in LRRK2 R1627P vs WT, LRRK2 −/− vs WT ( p value < 0.05, fold change >1.2), n = 3. b BP categories in GO enrichment analysis and KEGG pathway of DEGs in LRRK2 R1627P vs WT, n = 3. c BP categories in GO enrichment analysis and KEGG pathway of DEGs in LRRK2 −/− vs WT, n = 3. d Western blot analysis for TLR4, MyD88 and NF-κB protein levels in the small intestine of rats, n = 4. One-way ANOVA was used. e TEM images of rats small intestinal LP. Ec enterocytes, LP lamina propria. Scale bars 5 μm, n = 3. One-way ANOVA was used. f Western blot analysis for CD68 and CD163 protein levels in the small intestine of rats, n = 4. One-way ANOVA was used. g Immunofluorescence staining images of CD68 (green) and CD163 (red) (upper panel), IL-6 (green) and iNOS (red) (down panel) positive cells in the small intestinal LP of rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. One-way ANOVA was used. h Flow cytometry scatter plot of small intestianl LP isolated from rats, n = 3. CD11b + CD68 + CD163 - cells represent M1 population and CD11b + CD68 - CD163 + cells represent M2 population. i Statistical analysis of total macrophages, M1 macrophages and M2 macrophages in the small intestianl LP using flow cytometry. One-way ANOVA was used. j Immunofluorescence staining images of CD68 (green) and p S129 -α-Syn (red) positive cells in the small intestinal LP of rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. k Statistical analysis of immunofluorescence staining. One-way ANOVA was used. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: TAK-242 administration: TLR4 inhibitor TAK-242 was purchased from MedChemExpress (Monmouth Junction, NJ, USA), HY-11109.

Techniques: RNA Sequencing, Western Blot, Immunofluorescence, Staining, Flow Cytometry, Isolation

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a HE staining images of the small intestine of LPS-treated rats. The green circles in the lower panels highlight the red blood cells in the LP. Scale bar 100 μm, n = 4. b Images of goblet cells stained with AB-PAS in the small intestine of LPS-treated rats. Scale bar 100 μm, n = 4. c Immunohistochemical staining images of Cleaved caspase 3 in the small intestine of LPS-treated rats. Arrows show apoptotic IECs. Scale bar 100 μm, n = 4. d Western blot analysis for ZO-1, E-cadherin, TLR4, MyD88, NF-κB, CD68 and CD163 protein levels in the small intestine of LPS-treated rats, n = 4. e Immunofluorescence staining images and quantification of CD68/IL-6 (green) and CD163/iNOS/p S129 -α-Syn (red) positive cells in the small intestinal LP of LPS-treated rats. Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. One-way ANOVA was used. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: TAK-242 administration: TLR4 inhibitor TAK-242 was purchased from MedChemExpress (Monmouth Junction, NJ, USA), HY-11109.

Techniques: Staining, Immunohistochemical staining, Western Blot, Immunofluorescence

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a The representative small intestine images in rats treated with saline (control) or TLR4 inhibitor (TAK-242). Scale bar 5 cm, n = 8. b HE staining images of small intestine in rats treated with saline (control) or TLR4 inhibitor (TAK-242). Scale bar 100 μm, n = 4. c Measurement of small intestine length ( n = 8), villus height and crypt depth ( n = 4) in rats treated with saline (control) or TLR4 inhibitor (TAK-242). Two-way ANOVA was used; interaction p < 0.0001. d Immunohistochemical staining images of Ki67 in the small intestine, and the quantification of Ki67 positive cells in the crypts (determined based on the number of brown-stained cell nuclei in the crypts). Scale bar 100 μm, n = 4. Two-way ANOVA was used; interaction p < 0.05. e Images and quantification of goblet cells stained with AB-PAS in the small intestine. Scale bar 100 μm, n = 4. Two-way ANOVA was used; interaction p < 0.01. f Images and quantification of immunohistochemical staining for lysozyme in the crypt. Scale bar 100 μm, n = 4. Two-way ANOVA was used; interaction p < 0.01. g Western blot analysis for MUC2, Villin, ZO-1 and E-cadherin protein levels in the small intestine of WT, LRRK2 R1627P and LRRK2 −/− rats treated with saline (control) or TLR4 inhibitor (TAK-242), n = 4. One-way ANOVA was used. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: TAK-242 administration: TLR4 inhibitor TAK-242 was purchased from MedChemExpress (Monmouth Junction, NJ, USA), HY-11109.

Techniques: Saline, Control, Staining, Immunohistochemical staining, Western Blot

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: a , b Western blot analysis for TLR4, MyD88, NF-κB, CD68 and CD163 protein levels in the small intestine of WT, LRRK2 R1627P and LRRK2 −/− rats treated with saline (control) or TLR4 inhibitor (TAK-242), n = 4. One-way ANOVA was used. c Immunofluorescence staining images and quantification of CD68/IL-6 (green) and CD163/iNOS/p S129 -α-Syn (red) positive cells in the small intestinal LP of rats treated with saline (control) or TLR4 inhibitor (TAK-242). Nuclei were stained with DAPI (blue). Scale bar 100 μm, n = 4. Two-way ANOVA was used; for CD68/IL-6/iNOS analysis interaction p < 0.01; for CD163 analysis interaction p > 0.05; for p S129 -α-Syn analysis interaction p < 0.001. d GO enrichment analysis for DEGs in ConRP vs ConWT, TAKRP vs ConRP, TAKRP vs ConWT, n = 3. e Heatmap of DEGs from ConWT, ConRP and TAKRP group, n = 3. Data are presented as mean ± SEM, * p < 0.05, ** p < 0.01.

Article Snippet: TAK-242 administration: TLR4 inhibitor TAK-242 was purchased from MedChemExpress (Monmouth Junction, NJ, USA), HY-11109.

Techniques: Western Blot, Saline, Control, Immunofluorescence, Staining

Journal: NPJ Parkinson's Disease

Article Title: LRRK2 R1627P mutation amplifies environmental risk factors induced chronic inflammation and α-synuclein aggregation in the gut of rats

doi: 10.1038/s41531-026-01281-3

Figure Lengend Snippet: LRRK2 R1627P mutation significantly reduced intestianl endogenous total LRRK2, pT73-Rab10 protein level and the number of Paneth cells. Compared with age-macthed WT rats, LRRK2 R1627P rats showed shorter small intestine length, villus height, and crypt depth (from 8 months old); a significant decrease in the number of goblet cells, expression of barrier proteins, and microbial community diversity (from 16 months old); and a marked increase in the TLR4/MyD88/NF-κB signaling pathway, the number of M1 macrophages, the expression of p S129 -α-Syn, and the abundance of Lactobacillus (from 16 months old). Long-term administration of the TLR4 inhibitor TAK-242 effectively ameliorated the imbalance in intestinal homeostasis of LRRK2 R1627P rats.

Article Snippet: TAK-242 administration: TLR4 inhibitor TAK-242 was purchased from MedChemExpress (Monmouth Junction, NJ, USA), HY-11109.

Techniques: Mutagenesis, Expressing

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: Dependence of CIRP-induced chondrocyte damage on TLR4/NF-κB signaling. (A) Cytotoxicity of BAY 11-7082 and TAK-242 on chondrocytes at various concentration for 48 h, assessed using a CCK-8 assay. (B and C) Western blotting showing the protein expression of IκBα in chondrocytes and p65 in the nucleus of chondrocytes following treatment with the two inhibitors. (D and E) Western blotting showing the protein expression NLRP3, cleaved-caspase-1, ASC and IL-1β following treatment with the two inhibitors. (F and G) Western blotting showing the expression of extracellular matrix proteins in chondrocytes following treatment with the two inhibitors. n=3, * P<0.05, ** P<0.01, *** P<0.001. CIRP, cold-inducible RNA-binding protein; TLR4, Toll-like receptor 4; NLRP3, NLR family pyrin domain containing 3; Cle, cleaved.

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: Concentration Assay, CCK-8 Assay, Western Blot, Expressing, 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: 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