Journal: Frontiers in Pharmacology

Article Title: Enhanced therapeutic potential of paeoniflorin and vitamin B12 in intracerebropeduncle ethidium bromide-induced multiple sclerosis-like pathology

doi: 10.3389/fphar.2026.1792674

Figure Lengend Snippet: (A–D) PNN neuroprotective role in mitigating EBRO-induced alterations in levels of pro-inflammatory cytokines like TNF-α (A) , IL-1β (B) , IL-10 (C) in brain homogenates and TNF-α, IL-1β, and IL-10 in blood plasma (D) . To identify significant differences between groups, a one-way ANOVA and Tukey’s post hoc test were used for statistical analysis (A–D) . The statistical significance level was set at p < 0.01, and the data were displayed as mean ± standard deviation (SD). There were eight wistar rats (n = 8) in each experimental group. β v/s Sham Control, Vehicle Control, and PNN Perse; δ v/s EBRO; δα1 v/s EBRO + PNN50; δα2 v/s EBRO + PNN100, EBRO + PNN50; and δα3 v/s EBRO + VB12 (30), EBRO + PNN100, EBRO + PNN50.

Article Snippet: Also assessed neuroinflammatory cytokines TNF-alpha [KB1145; Krishgen Biosystem, Mumbai, India] and IL-1 Beta [KLR0119; Krishgen Biosystem, Mumbai, India] , and IL-10 [GENLISA, Krishgen, Maharashtra, India] ( ; ).

Techniques: Clinical Proteomics, Standard Deviation, Control

Journal: Advanced Science

Article Title: A Time‐Programmed Bilayer Wound Dressing for Dynamic Microenvironment Modulation and Full‐Thickness Regeneration in Diabetic Wounds

doi: 10.1002/advs.202512425

Figure Lengend Snippet: Polyphenols scavenge AGEs and inflammatory factors. Gene set enrichment analysis (GSEA) of (A) defense response, (B) negative regulation of IFN‐I mediated signaling pathway, (C) negative regulation of IL‐6, and (D) activation of innate immune response. Differences in (E) AGEs, (F) RAGE, (G) IL‐4, (H) IL‐6, (I) IL‐10, and (L) TNF‐α levels between groups Control, T2DM, and Polyphenol were detected by Elisa. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. n = 6 independent samples for each group, error bars represent mean ± SD.

Article Snippet: Mouse TNF alpha PicoKine ELISA Kit (catalog no. EK0527), Mouse IL‐10 PicoKine ELISA Kit (catalog no. EK0417), Mouse IL‐6 PicoKine ELISA Kit (catalog no. EK0411), Mouse IL‐4 PicoKine ELISA Kit (catalog no. EK0405) were purchased from BOSTER (Wuhan, China).

Techniques: Activation Assay, Control, Enzyme-linked Immunosorbent Assay

Journal: Advanced Science

Article Title: A Time‐Programmed Bilayer Wound Dressing for Dynamic Microenvironment Modulation and Full‐Thickness Regeneration in Diabetic Wounds

doi: 10.1002/advs.202512425

Figure Lengend Snippet: Expression of inflammatory and anti‐inflammatory factors during wound healing. (A) Representative images of immunohistochemical staining of IL‐4, (B) IL‐10, (C) TNF‐α, (D) IL‐6 in Control, Nanofiber, Polyhenol, PDGF‐BB, and Polyhenol+PDGF‐BB groups at 3 and 7 days. (E–H) Quantification of IL‐4, IL‐10, TNF‐α, and IL‐6. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: Mouse TNF alpha PicoKine ELISA Kit (catalog no. EK0527), Mouse IL‐10 PicoKine ELISA Kit (catalog no. EK0417), Mouse IL‐6 PicoKine ELISA Kit (catalog no. EK0411), Mouse IL‐4 PicoKine ELISA Kit (catalog no. EK0405) were purchased from BOSTER (Wuhan, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Control

Journal: Advanced Science

Article Title: Loss of SOCS1 in Donor T Cells Exacerbates Intestinal GVHD by Driving a Chemokine‐Dependent Pro‐Inflammatory Immune Microenvironment

doi: 10.1002/advs.202513735

Figure Lengend Snippet: Socs1 deficiency in T cells drives effector differentiation and enhances inflammatory responses in CD8 + T cells. (A) Experimental schematic. Splenocytes from WT (littermate control; Socs1 fl/fl ) and cKO (LckCre‐ Socs1 fl/fl ) mice were isolated and sorted by FACS for CD45 + cells and subjected to single‐cell RNA sequencing (scRNA‐seq, n = 5 per group). Alternatively, CD8 + T cells from the spleen of WT and cKO mice were sorted by FACS and subjected to bulk RNA‐seq, ATAC‐seq, and CUT&Tag analyses. (B) UMAP plot of 105 040 single cells from CD45 + splenocytes colored by annotated immune cell subsets. (C) UMAP visualization of CD45 + splenocytes, split by origin. (D) Comparison of the proportions of celltypes between WT and cKO groups. (E) UMAP plot of 19 490 T cells extracted from Figure and colored by annotated T cell subsets. (F) UMAP visualization of T cells, split by origin. (G) Heatmap of Ro/e (Ratio of observed to expected) scores for T cell subtypes in WT and cKO mice. The scores, calculated from scRNA‐seq cell counts, indicate the relative enrichment (red, Ro/e > 1) or depletion (white/light orange, Ro/e < 1) of each population within each genotype. Numerical values are presented alongside a semi‐quantitative summary. (H) UMAP plot of 9159 CD8 + T cells extracted from Figure and colored by annotated T cell subsets. (I) Comparison of the proportions of indicated CD8 + T cell clusters between WT and cKO groups. (J) Representative flow cytometry plots and frequencies of naive T cells (Tn; CD44 − CD62L + ), central memory (Tcm; CD44 + CD62L + ), and effector memory (Tem; CD44 + CD62L − ) in CD8 + T cells from peripheral blood (PB, left panel) and spleen (SP, right panel) (n = 5 per group). (K) Bar plots showing the expression of perforin, GZMB, TNF‐α, IFN‐γ, IL‐2, and CD107a in CD8 + T cells from WT and cKO mice, as measured by flow cytometry (n=5 per group). (L) UMAP visualization of integrated T‐cell transcriptomes from WT (left, 25 304 cells) and cKO (right, 21 588 cells) groups. Each point represents a single cell, colored by the frequency of its corresponding TCR clonotype, highlighting clonally expanded cells. (M) Quantification of overall TCR repertoire diversity. The Gini index (top) and Shannon entropy (bottom) were calculated for the entire T‐cell population from each mouse. (N) Distribution and clonal size of T cells across identified subsets. Barplot showing the absolute cell counts (left panels) and the clonal size composition (right panels) for each T‐cell subset from WT and cKO mice. Data represent one experiment out of two independent experiments. P values were determined using two‐sided Wilcoxon rank‐sum test (D, I, M) or unpaired two‐tailed Student's t‐test (J‐K). Data represent mean ± SEM (D, I, M) or mean ± SD (J‐K). ∗ p <.05, ∗∗ p <.01 and ∗∗∗∗ p <.0001.

Article Snippet: The following kits were used: IL‐1β (Proteintech, KE10003), IL‐6 (Proteintech, KE10007), and TNF‐α (Proteintech, KE10002).

Techniques: Control, Isolation, Single Cell, RNA Sequencing, Comparison, Flow Cytometry, Expressing, Two Tailed Test

Journal: Advanced Science

Article Title: Loss of SOCS1 in Donor T Cells Exacerbates Intestinal GVHD by Driving a Chemokine‐Dependent Pro‐Inflammatory Immune Microenvironment

doi: 10.1002/advs.202513735

Figure Lengend Snippet: Evolution of small intestinal immune cell composition following transplantation of Socs1 cKO CD8 + T cells. (A) Experimental schematic. Lethally irradiated BALB/c recipient mice were transplanted with splenic T cells from WT mice (WT group) or cKO mice (cKO group), along with 5 × 10 6 TCD‐BM cells from WT mice. Survival was monitored daily. Body weight and GVHD score were assessed every five days. Immune cells in PB and small intestine from WT and cKO groups were assessed on Day 7, Day 16, and Day 24 by flow cytometry. (B) Survival analysis of recipients transplanted with 1 × 10 6 , 2 × 10 6, or 3 × 10 6 splenic T cells from WT or cKO mice (n = 10 mice per group). A control group received TCD‐BM only. Median survival times for cKO groups were 36 (1 × 10 6 ), 33 (2 × 10 6 ), and 26 days (3 × 10 6 ), respectively. In the corresponding WT groups, 9/10 mice in the 1 × 10 6 group survived to the end of the observation period, with median survival times of 35 days (2 × 10 6 ), and 42.5 days (3 × 10 6 ). Data were pooled from two independent experiments (n=10 mice/group). (C‐D) GVHD score (C) and body weight changes (D) in WT and cKO recipients transplanted with 1 × 10 6 splenic T cells. (E) Proportions of CD8 + T cells and monocytes in PB, and proportions of CD8 + T cells in the IEL and LP on Day 7, 16, and 24 post‐transplantation. (F) Boxplots showing the expression of TNF‐α, IFN‐γ, perforin, and CD107a in CD8 + T cells in IEL by flow cytometry on Day 7 and 16 post‐transplantation. (G) Experimental schematic of scRNA‐seq. Lymphocytes from IEL and LP were isolated and collected from 3‐4 mice, followed by FACS sorting for CD45 + donor‐derived (H‐2 b+ H‐2 d− ) cells respectively. Sorted cells from IEL and LP were mixed in a 1:2 ratio and subsequently subjected to scRNA‐seq. (H‐I) UMAP plot of all immune cells colored by samples (H) or annotated subsets (I). (J) UMAP plot showing annotated CD8 + T cell subsets (left), with barplot illustrating the proportion of each annotated subset in recipient intestines at indicated time points between WT and cKO groups (right). (K) Violin plots comparing the expression of cytotoxic molecules and cytokine receptor genes in CD8 + T cells on Day 26 post‐transplantation between WT and cKO groups. (L) UMAP plot showing annotated myeloid cell subsets (left), with barplot illustrating the proportion of each annotated subset in recipient intestines at indicated time points between WT and cKO groups (right). (M) Violin plots comparing expression of M1‐ and M2‐associated marker genes in myeloid cells on Day 26 post‐transplantation between WT and cKO groups. Data represent three independent experiments. P values were determined using chi‐squared test (B) or unpaired two‐tailed Student's t‐test (E, F) or two‐sided Wilcoxon rank‐sum test (K, M). Differences in GVHD scores and body weight between the WT and cKO groups at each time point were analyzed using multiple unpaired two‐tailed Student's t ‐tests (C, D). Data represent mean ± SEM (C‐E). ∗ p <.05, ∗∗ p <.01, ∗∗∗ p <.001 and ∗∗∗∗ p <.0001.

Article Snippet: The following kits were used: IL‐1β (Proteintech, KE10003), IL‐6 (Proteintech, KE10007), and TNF‐α (Proteintech, KE10002).

Techniques: Transplantation Assay, Irradiation, Flow Cytometry, Control, Expressing, Isolation, Derivative Assay, Marker, Two Tailed Test

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