Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Identification of TLR ‐7–expressing cells in SMG s from patients with IgG4‐ RD and in normal secondary lymphoid organs from patients with oral squamous cell carcinoma. A , Staining of serial sections of normal tonsil, normal lymph node, and IgG4‐ RD SMG s with H&E, and for TLR ‐7, CD 68 as a marker of both M1 and M2 macrophages, CD 163 as a marker of M2 macrophages, CD 11c as a marker of myeloid dendritic cells, and CD 123 as a marker of plasmacytoid dendritic cells. Outlined areas indicate GC s. Mayer's hematoxylin (blue) counterstained; bars = 100 μm. B , Double immunostaining for TLR ‐7 (red), CD 123 (green), and CD 163 (green) in normal tonsil and SMG s from representative patients with IgG4‐ RD . DAPI (blue) counterstained; bars = 50 μm. See Figure for definitions.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques: Expressing, Staining, Marker, Double Immunostaining

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Expression of interleukin‐33 ( IL ‐33) and candidate TLR s in SG s from patients with IgG4‐ RD . A , Expression levels of mRNA for IL ‐33 in SG s from healthy controls (n = 10), patients with chronic sialadenitis (n = 10), patients with SS (n = 15), and patients with IgG4‐ RD (n = 15). B , Distribution of IL ‐33 in SG s from a representative healthy control, patient with chronic sialadenitis, patient with SS , and patient with IgG4‐ RD . Mayer's hematoxylin (blue) counterstained; bars = 100 μm. C , Correlation between expression levels of IL ‐33 mRNA and candidate TLR s in SG s from patients with IgG4‐ RD (n = 15), as determined by Spearman's rank correlation test. D , Schematic illustration of the extraction of CD 163+ M2 macrophages stimulated with TLR ‐7 agonist R848. Cells were cultivated as described in Patients and Methods. PBMC = peripheral blood mononuclear cell. E , Production of IL ‐33 by CD 163+ M2 macrophages stimulated with R848, as determined by enzyme‐linked immunosorbent assay. IL ‐33 levels increased in a concentration‐dependent manner. In A and E , bars show the mean ± SD . * = P < 0.05; ** = P < 0.01 by Kruskal‐Wallis test. See Figure for other definitions.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques: Expressing, Control, Extraction, Enzyme-linked Immunosorbent Assay, Concentration Assay

Journal: Arthritis & Rheumatology (Hoboken, N.j.)

Article Title: Activated M2 Macrophages Contribute to the Pathogenesis of IgG4‐Related Disease via Toll‐like Receptor 7/Interleukin‐33 Signaling

doi: 10.1002/art.41052

Figure Lengend Snippet: Schematic model of the role of Toll‐like receptor 7 ( TLR ‐7)–positive M2 macrophages in the initiation of Ig4‐related disease. TLR ‐7 expressed on M2 macrophages recognize some RNA viruses or self RNA from apoptotic cells. Activated M2 macrophages secrete interleukin‐33 ( IL ‐33) and promote production of Th2 cytokines that lead to IgG4 class‐switching and fibrosis. TGF β = transforming growth factor β.

Article Snippet: Cells (5 × 10 6 cells/ml) were cultured using a CellXVivo Human M2 Macrophage Differentiation Kit (R&D Systems) for 6 days in a humidified chamber with 5% CO 2 at 37°C.

Techniques:

Journal: Chinese medicine

Article Title: Shen-Bai-Jie-Du decoction suppresses the progression of colorectal adenoma to carcinoma through regulating gut microbiota and short-chain fatty acids.

doi: 10.1186/s13020-024-01019-4

Figure Lengend Snippet: Fig. 9 The schematic diagram of the mechanism of SBJDD. SBJDD may exert its inhibitory effects on colorectal adenoma carcinogenesis by regulating gut microbiota, promoting the production of SCFAs, activating G protein-coupled receptors GPR43, GPR41 and GPR109a, inhibiting histone deacetylases HDAC1 and HDAC3, inducing M2-like macrophage polarization, reducing intestinal inflammation, inhibiting cancer cell proliferation and restoring intestinal barrier function

Article Snippet: Anti-GAPDH (no. 2118S), anti-β-Tubulin (no. 2146S) and Mouse Reactive M1 vs M2 Macrophage IHC Antibody Sampler Kit (no. 97624 T) were obtained from Cell Signaling Technology (Beverly, USA).

Techniques:

Journal: Genes & Diseases

Article Title: The miR-21-5p enriched in the apoptotic bodies of M2 macrophage-derived extracellular vesicles alleviates osteoarthritis by changing macrophage phenotype

doi: 10.1016/j.gendis.2022.09.010

Figure Lengend Snippet: Establishment of M1 and M2 Mφs and characterization of Mφ-ABs. (A) Schematic illustration of apoptotic bodies-guided macrophage reprogramming. M2 Mφ-derived apoptotic bodies (M2-ABs) can trigger the switch of M1 to M2 Mφs transformation and alleviate the progression of osteoarthritis. (B) Schematic diagram of obtaining M1 and M2 Mφs. (C) Western blot analysis showed the expression of Mφ markers 24 h after polarization. (D) Representative SEM image of ABs. Scale bar, 500 nm. (E) Size distribution of ABs measured by DLS. (F) Western blot analysis specific protein markers of Mφs and Mφs-ABs. (G) Confocal images of M1 Mφs incubation with 10, 25, 50, and 100 μg/mL of DiD-labeled M2-ABs in 4 or 6 h, respectively (Red: DiD-labeled M2-ABs; Blue: cell nuclear). Scale bars, 50 μm. (H) Relative fluorescence intensity of DiD-labeled M2-ABs internalized in M1 Mφs.

Article Snippet: The cytotoxicity of ABs with M1 and M2 Mφs were evaluated using Cell Counting Kit-8 (CCK-8, Beyotime Bio-Tech, China).

Techniques: Derivative Assay, Transformation Assay, Western Blot, Expressing, Incubation, Labeling, Fluorescence

Journal: Genes & Diseases

Article Title: The miR-21-5p enriched in the apoptotic bodies of M2 macrophage-derived extracellular vesicles alleviates osteoarthritis by changing macrophage phenotype

doi: 10.1016/j.gendis.2022.09.010

Figure Lengend Snippet: M2-ABs guided reprogramming of M1 Mφs to M2 Mφs. (A) Immunostaining of iNOS and Arginase in M1 Mφs after 24 h, 48 h and 72 h incubation with 50 μg/mL of M2-ABs, respectively. (B) Western blot analysis of M1 Mφs treated with 50 μg/mL of M2-ABs over time. (C) Relative gray value of M1 and M2 Western blot markers in Mφs. (D) Heat map for expression analysis (using RT-qPCR) of M1 and M2 macrophages marker genes ( n = 3). (E) FACS histogram showed reprogramming efficiency of M1 Mφs treated with 50 μg/mL of M2-ABs over time.

Article Snippet: The cytotoxicity of ABs with M1 and M2 Mφs were evaluated using Cell Counting Kit-8 (CCK-8, Beyotime Bio-Tech, China).

Techniques: Immunostaining, Incubation, Western Blot, Expressing, Quantitative RT-PCR, Marker

Journal: Genes & Diseases

Article Title: The miR-21-5p enriched in the apoptotic bodies of M2 macrophage-derived extracellular vesicles alleviates osteoarthritis by changing macrophage phenotype

doi: 10.1016/j.gendis.2022.09.010

Figure Lengend Snippet: In vitro anti-inflammatory and chondrocyte protective effects of RM2 Mφs. (A) The concentrations of macrophage serum-free medium cytokines were determined using the Bio-Plex mouse cytokine 23-Plex panel. (B) a . Schematic diagram of RM2 co-culture with chondrocytes; b. EdU proliferation assays of chondrocytes exposed to classically activated M1-Mφs, alternatively activated M2-Mφs and RM2 for 24 h were performed; c. EdU positive fluorescence intensity analysis. ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The cytotoxicity of ABs with M1 and M2 Mφs were evaluated using Cell Counting Kit-8 (CCK-8, Beyotime Bio-Tech, China).

Techniques: In Vitro, Co-Culture Assay, Fluorescence

Journal: Genes & Diseases

Article Title: The miR-21-5p enriched in the apoptotic bodies of M2 macrophage-derived extracellular vesicles alleviates osteoarthritis by changing macrophage phenotype

doi: 10.1016/j.gendis.2022.09.010

Figure Lengend Snippet: Micro-RNA sequencing (miRNA-seq) analysis and molecular mechanism of M2-ABs. (A) Top 50 known miRNAs that were detected in M2-ABs. (B) Co-localization of macrophages and miR-21a-5p. (C) Western blot analysis of the effect of miR-21a-5p inhibitors on M2-ABs guided M1-Mφs reprogramming to M2-Mφs. (D) Flow cytometry (FACS) histogram shows the effect of miR-21a-5p inhibitors on M2-ABs guided M1-Mφs reprogramming to M2-Mφs.

Article Snippet: The cytotoxicity of ABs with M1 and M2 Mφs were evaluated using Cell Counting Kit-8 (CCK-8, Beyotime Bio-Tech, China).

Techniques: RNA Sequencing, Western Blot, Flow Cytometry

Journal: Genes & Diseases

Article Title: The miR-21-5p enriched in the apoptotic bodies of M2 macrophage-derived extracellular vesicles alleviates osteoarthritis by changing macrophage phenotype

doi: 10.1016/j.gendis.2022.09.010

Figure Lengend Snippet: In vivo biodistribution of M2-ABs. (A) Real-time in vivo imaging of Cy7 NHS labeled M2-ABs. The mice were analyzed at the indicated times after knee injection of phosphate-buffered saline (PBS) and 50 μg/μl of M2-ABs. (B) Ex vivo imaging of major organs at day 3 after mice had been treated with M2-ABs. (C) Viscera distribution of fluorescent at day 3 after knee injection of Cy7-NHS labeled M2-ABs.

Article Snippet: The cytotoxicity of ABs with M1 and M2 Mφs were evaluated using Cell Counting Kit-8 (CCK-8, Beyotime Bio-Tech, China).

Techniques: In Vivo, In Vivo Imaging, Labeling, Injection, Saline, Ex Vivo, Imaging

Journal: Genes & Diseases

Article Title: The miR-21-5p enriched in the apoptotic bodies of M2 macrophage-derived extracellular vesicles alleviates osteoarthritis by changing macrophage phenotype

doi: 10.1016/j.gendis.2022.09.010

Figure Lengend Snippet: μCT evaluations of M2-ABs treated OA induced by ACLT. (A) Three-dimensional μCT images of frontal views of the knee joints at 5 weeks after sham operation or ACLT operation. (B) Sagittal views of medial compartment subchondral bone. ( C) Quantitative analysis of BV, BV/TV, Tb.Sp, Tb. n and Tb.Th. ∗ P < 0.05, ∗∗ P < 0.01. Scale = 1 mm.

Article Snippet: The cytotoxicity of ABs with M1 and M2 Mφs were evaluated using Cell Counting Kit-8 (CCK-8, Beyotime Bio-Tech, China).

Techniques:

Journal: Genes & Diseases

Article Title: The miR-21-5p enriched in the apoptotic bodies of M2 macrophage-derived extracellular vesicles alleviates osteoarthritis by changing macrophage phenotype

doi: 10.1016/j.gendis.2022.09.010

Figure Lengend Snippet: M2-ABs alleviated the disease progression of OA in vivo ( n = 5 for each group). (A) Representative images of H&E staining and (B) Safranin O-fast green staining from each group. (C) Representative immunostaining images of M2 (Arginase, red) and (D) M1 (iNOS, green) Mφ in synovial tissues. (E) Depth of cartilage. (F) Total Mankin scores of articular cartilage. (G) Fluorescence intensity analysis of CD206 and (H) CD86 in synovial tissue. ∗∗∗ P < 0.001, ∗∗ P < 0.01, ∗ P < 0.05.

Article Snippet: The cytotoxicity of ABs with M1 and M2 Mφs were evaluated using Cell Counting Kit-8 (CCK-8, Beyotime Bio-Tech, China).

Techniques: Biomarker Discovery, In Vivo, Staining, Immunostaining, Fluorescence