Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: OXA activated TLR4 signaling in ESCC cells. The cells were treated by OXA (25 µM) for 24 h. A , B . The mRNA expressions of TLR4 and MYD88 in ESCC cell lines and normal esophageal cells were detected by qRT-PCR. C . ICC showed the immunocytochemical activity of NF-κB p65, p-NF-κB p65, COX-2, and MYD88. Scale bar: 50 μm. D . qRT-PCR analysis of mRNA levels of IL-1β, IL-6, COX-2, CXCL5, and CXCL8. *p < 0.05, **p < 0.01, ***p < 0.001 vs. Control group

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Quantitative RT-PCR, Activity Assay, Control

Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: TLR4 knockout enhances the sensitivity of OXA chemotherapy in ESCC in vivo. A . Number of tumors. B . HE staining of esophageal epithelial tissues. Scale bar: 50 μm. C . Body weight of mice. D , E . the levels of serum IL-1β and IL-6 in mice from different groups were detected by ELISA. F . The immunohistochemical activities of PCNA, CK14, Cyclin D1, COX-2, S100A8 and S100A9 in the esophageal tissue were detected. Scale bar: 100 μm. G , H . The mRNA levels of inflammatory cytokines and glycolysis-related proteins in esophageal tissue were detected by qRT-PCR. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. WT group; # p < 0.05, ## p < 0.01, ### p < 0.001 vs. WT + 4NQO + OXA group

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Knock-Out, In Vivo, Staining, Enzyme-linked Immunosorbent Assay, Immunohistochemical staining, Quantitative RT-PCR

Journal: BMC Gastroenterology

Article Title: Inhibition of TLR4 enhances oxaliplatin chemotherapy sensitivity in esophageal squamous cell carcinoma by suppressing inflammation and glycolysis

doi: 10.1186/s12876-026-04663-2

Figure Lengend Snippet: Schematic diagram illustrating the mechanism by which TLR4 inhibition enhances oxaliplatin (OXA) chemosensitivity in esophageal squamous cell carcinoma (ESCC). OXA treatment upregulates TLR4 and its downstream adaptor protein MYD88, which activates the phosphorylation of NF-κB p65. This activation drives two parallel pathways: (1) the inflammatory response, characterized by the upregulation of pro-inflammatory factors such as IL-6, COX-2, and CXCL5; (2) the glycolytic metabolic reprogramming, mediated by the HIF-1α/GLUT1 axis and enhanced expression of glycolytic enzymes including PFKM and LDHB. These two pathways synergistically promote ESCC cell proliferation, migration, and invasion, ultimately reducing OXA chemosensitivity. Inhibition of TLR4 (via genetic knockout, shRNA knockdown, or pharmacological inhibitor TAK-242) or its downstream mediator MYD88 (via shRNA knockdown or inhibitor ST2825) blocks NF-κB p65 phosphorylation, thereby suppressing both the inflammatory response and glycolytic activity. This dual inhibition disrupts the adaptive survival mechanisms of ESCC cells, potentiating the anti-tumor efficacy of OXA

Article Snippet: Serum concentrations of IL-6 (JL20268, Jonin) and IL-1β (KE10003, Proteintech) were quantified using commercial ELISA kits according to manufacturers’ protocols.

Techniques: Inhibition, Phospho-proteomics, Activation Assay, Expressing, Migration, Knock-Out, shRNA, Knockdown, Activity Assay

Journal: Frontiers in Immunology

Article Title: Trem2 activation by renal tubular debris sustains Arg1 + macrophage survival and promotes tubular epithelial repair in renal ischemia–reperfusion injury

doi: 10.3389/fimmu.2026.1819941

Figure Lengend Snippet: Tubular cell debris triggers Trem2 upregulation and stimulates proliferation in Arg1 + macrophages. (A, B) IL-4 treatment of RAW264.7 cells for 24 h significantly increased Arg1 transcription, Arg1 + macrophage proportion, and intracellular Arg1 protein intensity ( n = 3-6). (C, D) To mimic the IRI microenvironment, freeze-thaw–induced tubular cell debris were co-cultured with IL-4–pretreated RAW264.7 cells ( n = 6). This induced robust upregulation of Trem2 , Spp1 , and Apoe transcripts. (E) Tubular cell debris increased both the number and proliferative activity of Arg1 + macrophages. Higher debris concentrations further increased both measures, suggesting proliferation scales with debris exposure ( n = 6). (F, G, H) Flow cytometry revealed increased Trem2 receptor intensity on Arg1 + macrophages and a higher proportion of Trem2 + Arg1 + macrophages after debris stimulation ( n = 6). (I) Levels of Spp1 and Apoe in culture supernatants were significantly elevated following debris treatment ( n = 8). (J) Schematic illustration of the experimental design. Mouse primary BMDMs were pretreated with IL-4 to induce differentiation toward an Arg1 high phenotype, followed by co-culture with renal tubular debris. (K) Western blot analysis showed that IL-4 stimulation markedly upregulated Arg1 protein expression in BMDMs ( n = 6). (L, M) RT-qPCR and Western blot analyses confirmed that renal tubular debris further induced the transcriptional and translational upregulation of Trem2 in Arg1 high BMDMs ( n = 4). (N) Renal tubular debris promoted the viability and proliferation of Arg1 high BMDMs in a concentration-dependent manner ( n = 6). Significance was evaluated using Student’s unpaired t test and one-way ANOVA followed by Tukey’s test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns , no significance.

Article Snippet: RAW264.7 cells were treated with 20 ng/ml IL-4 protein (MedChemExpress, HY- P70653 ) for 24 hours to induce polarization.

Techniques: Cell Culture, Activity Assay, Flow Cytometry, Co-Culture Assay, Western Blot, Expressing, Quantitative RT-PCR, Concentration Assay

Journal: Frontiers in Immunology

Article Title: Trem2 activation by renal tubular debris sustains Arg1 + macrophage survival and promotes tubular epithelial repair in renal ischemia–reperfusion injury

doi: 10.3389/fimmu.2026.1819941

Figure Lengend Snippet: Trem2 is essential for the survival and repair function of Arg1 + macrophages. (A–D) Establishment of stable Trem2 knockdown (KD) RAW264.7 cells using shRNA lentiviral transduction, confirmed by GFP fluorescence, qPCR, and Western blot ( n = 3-6). (E) Schematic illustration of the co-culture system of IL-4–pretreated Trem2 KD RAW264.7 cells with tubular cell debris. (F) Trem2 deficiency markedly impaired debris-induced proliferation of Arg1 + macrophages across both low and high debris concentrations ( n = 6). (G) Debris-induced expansion of Arg1 + macrophages was significantly reduced upon Trem2 Knockdown ( n = 6). (H, I) Apoptosis assays showed increased apoptosis of Arg1 + macrophages under Trem2 Knockdown ( n = 3). (J, K) Trem2 Knockdown attenuated IL-4–induced Arg1 expression, suggesting impaired polarization toward a pro-repair phenotype ( n = 3-6). (L–N) Conditioned medium from Trem2-sufficient Arg1 + macrophages promoted TCMK-1 cell proliferation and expansion, whereas Trem2 KD abolished this pro-regenerative effect ( n = 8). (O) Levels of spermidine and spermine in the supernatants of Arg1 + macrophages were increased upon stimulation with tubular debris and reduced by Trem2 knockdown ( n = 8). (P) Tubular debris stimulation increased HGF and VEGF levels in Arg1 + macrophage supernatants, which were reduced by Trem2 knockdown, while IL-10 levels remained unchanged ( n = 8). Significance was evaluated using Student’s unpaired t test, one-way ANOVA, or two-way ANOVA followed by Tukey’s test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns , no significance.

Article Snippet: RAW264.7 cells were treated with 20 ng/ml IL-4 protein (MedChemExpress, HY- P70653 ) for 24 hours to induce polarization.

Techniques: Knockdown, shRNA, Transduction, Fluorescence, Western Blot, Co-Culture Assay, Expressing

Journal: Frontiers in Immunology

Article Title: Trem2 activation by renal tubular debris sustains Arg1 + macrophage survival and promotes tubular epithelial repair in renal ischemia–reperfusion injury

doi: 10.3389/fimmu.2026.1819941

Figure Lengend Snippet: Inhibition of Trem2 reduces the survival of Arg1 high BMDMs and impairs their ability to promote renal tubular epithelial cell proliferation. (A) Schematic illustration of the experimental design: BMDMs were treated with TREM2-IN-1 in combination with IL-4 and subsequently co-cultured with renal tubular cell debris. (B) Western blot analysis confirmed that TREM2-IN-1 markedly downregulated Trem2 protein levels in Arg1 high BMDMs ( n = 6). (C) Tubular cell debris enhanced the viability of Arg1 high BMDMs, whereas TREM2-IN-1 treatment significantly reduced both BMDMs viability and BMDMs number ( n = 6). (D) Flow cytometry revealed that inhibition of Trem2 significantly decreased the survival rate of Arg1 high BMDMs and markedly increased apoptosis ( n = 3). (E) Schematic of the conditioned medium experiment: Culture supernatants were collected from Arg1 high BMDMs and applied to TCMK-1 cells. (F) Conditioned medium from BMDMs treated with IL-4 and renal tubular cell debris significantly promoted TCMK-1 cell proliferation, whereas the addition of TREM2-IN-1 markedly attenuated this pro-proliferative effect ( n = 6). (G) Measurement of Arg1 high BMDM-derived secreted factors: TREM2-IN-1 treatment significantly reduced the levels of spermidine, spermine, HGF, and VEGF in the conditioned medium, while IL-10 levels remained unchanged ( n = 6). Significance was evaluated using Student’s unpaired t test, one-way ANOVA, or two-way ANOVA followed by Tukey’s test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns , no significance.

Article Snippet: RAW264.7 cells were treated with 20 ng/ml IL-4 protein (MedChemExpress, HY- P70653 ) for 24 hours to induce polarization.

Techniques: Inhibition, Cell Culture, Western Blot, Flow Cytometry, Derivative Assay

Journal: Frontiers in Immunology

Article Title: Trem2 activation by renal tubular debris sustains Arg1 + macrophage survival and promotes tubular epithelial repair in renal ischemia–reperfusion injury

doi: 10.3389/fimmu.2026.1819941

Figure Lengend Snippet: Inhibition of Ttem2 reduces the viability of Arg1 high BMDMs by upregulating Pten and suppressing Bcl2. (A) Schematic illustration of the experimental design: IL-4–induced Arg1 high BMDMs were first treated with TREM2-IN-1 to inhibit Trem2, followed by treatment with the PTEN inhibitor VO-Ohpic, and subsequently co-cultured with renal tubular cell debris. (B, C) Western blot analysis showed that, compared with the control group, VO-Ohpic treatment significantly downregulated Pten protein expression in Arg1 high BMDMs and markedly upregulated the expression of the key anti-apoptotic protein Bcl2 ( n = 6). (D) VO-Ohpic treatment effectively reversed the TREM2-IN-1–induced reduction in cell viability and cell number of Arg1 high BMDMs ( n = 6). (E) Proposed mechanism: Tubular cell debris generated during IRI, in conjunction with Apoe released by Arg1 + macrophages, activates Trem2 and further upregulates its expression. Elevated Trem2 signaling suppresses Pten and upregulates the anti-apoptotic factor Bcl2, thereby promoting debris clearance, Arg1 + macrophage survival. Surviving Arg1 + macrophages release spermidine, spermine, HGF, and VEGF, which enhance renal tubular epithelial cell regeneration and repair. Significance was evaluated using Student’s unpaired t test and two-way ANOVA followed by Tukey’s test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001, ns , no significance.

Article Snippet: RAW264.7 cells were treated with 20 ng/ml IL-4 protein (MedChemExpress, HY- P70653 ) for 24 hours to induce polarization.

Techniques: Inhibition, Cell Culture, Western Blot, Control, Expressing, Generated