Journal: Bone Research

Article Title: FABP4 secreted by M1-polarized macrophages promotes synovitis and angiogenesis to exacerbate rheumatoid arthritis

doi: 10.1038/s41413-022-00211-2

Figure Lengend Snippet: The expression of FABP4 in the RA mouse model, BMDMs and RA patients. a , b FABP4 concentrations in the synovial fluid ( a ) and serum ( b ) of controls and RA patients ( n = 8 per group) were assessed by ELISA. C Representative images and quantification of HE staining and FABP4 immunohistochemical staining in the synovium of control and C57BL/6 J mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bars: 50 μm, 100 μm and 200 μm. d Representative images and quantitative analysis of coimmunostaining of FABP4 with F4/80 or NOS2 in the synovium of control and C57BL/6 J mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bars: 12.5 μm. e Representative images and quantification of safranin O and fast green staining and FABP4 immunohistochemical staining in knee cartilage from controls and C57BL/6 J mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bars: 50 μm and 100 μm. f Western blot showing FABP4 and NOS2 in BMDMs stimulated with 50, 200, or 500 ng·mL −1 LPS for 12 h or 24 h. g Quantitative PCR analysis of NOS2, Arg-1, and FABP4 mRNA expression in BMDMs ( n = 3 per group). h FABP4 concentrations in the supernatant of BMDMs stimulated with 50, 200, or 500 ng·mL −1 LPS for 12 h or 24 h ( n = 3 per group) were assessed by ELISA. Student’s t -test or one-way analysis of variance (ANOVA) and Tukey’s multiple comparison test. * P < 0.05, ** P < 0.01; ns, no significance. The data are shown as the mean ± SEM

Article Snippet: We used human and mouse FABP4 ELISA kits (Elabscience Biotechnology, Bethesda, MD, USA: #E-EL-H0285c and #E-EL-M2404c) to analyze the level of FABP4 in the supernatant of macrophages stimulated with lipopolysaccharide (LPS), the serum of C57BL/6 J and TSC1KO mice, and human serum and synovial fluid.

Techniques: Expressing, Enzyme-linked Immunosorbent Assay, Staining, Immunohistochemical staining, Control, Western Blot, Real-time Polymerase Chain Reaction, Comparison

Journal: Bone Research

Article Title: FABP4 secreted by M1-polarized macrophages promotes synovitis and angiogenesis to exacerbate rheumatoid arthritis

doi: 10.1038/s41413-022-00211-2

Figure Lengend Snippet: The effect of FABP4 and M1-polarized macrophage supernatant on HUVECs, FLSs, and chondrocytes in vitro. a , b Tube formation assay and quantification of HUVECs cultured with vehicle (C), rhFABP4 (P), rhFABP4 + BMS309403 (PB), M1-polarized macrophage supernatant (S), or M1-polarized macrophage supernatant+BMS309403 (SB) ( n = 3 per group). Scale bar: 100 µm. c Immunoblot analysis of FABP4 and VEGFα in HUVECs cultured with C, P, PB, S, or SB for 24 h. d , g Representative images and quantification of BrdU (green) immunofluorescence ( d ) and Transwell assays ( g ) in HUVECs and FLSs treated with C, P, PB, S, or SB for 24 h ( n = 3 per group). Scale bar: 25 µm, 100 µm. e , f Immunoblot analysis of ERK1/2, p-ERK1/2, P65, and p-P65 in HUVECs ( e ) and FLSs ( f ) treated with C, P, PB, S, or SB for 1 h. h Toluidine blue staining of ATDC5 cells treated with C, P, PB, S, or SB for 24 h ( n = 3 per group). i Immunoblot analysis of FABP4, MMP13, Sox9, and Col2a1 in primary chondrocytes treated with C, P, PB, S, or SB for 24 h. j Immunoblot analysis of P65 and p-P65 in primary chondrocytes treated with C, P, PB, S, or SB for 1 h. One-way ANOVA and Tukey’s multiple comparison test. * P < 0.05, ** P < 0.01; # P < 0.05, ## P < 0.01 compared to the control. The data are shown as the mean ± SEM

Article Snippet: We used human and mouse FABP4 ELISA kits (Elabscience Biotechnology, Bethesda, MD, USA: #E-EL-H0285c and #E-EL-M2404c) to analyze the level of FABP4 in the supernatant of macrophages stimulated with lipopolysaccharide (LPS), the serum of C57BL/6 J and TSC1KO mice, and human serum and synovial fluid.

Techniques: In Vitro, Tube Formation Assay, Cell Culture, Western Blot, Immunofluorescence, Staining, Comparison, Control

Journal: Bone Research

Article Title: FABP4 secreted by M1-polarized macrophages promotes synovitis and angiogenesis to exacerbate rheumatoid arthritis

doi: 10.1038/s41413-022-00211-2

Figure Lengend Snippet: Recombinant FABP4 exacerbates the development of RA in C57BL/6 J mice. a Representative images and quantitative analysis of FABP4 were assessed by immunohistochemical staining and coimmunostaining of FABP4 with F4/80 or NOS2 in the synovium of control and RA mice treated with vehicle or rmFABP4 for 4 and 8 weeks ( n = 10 per group). Scale bars: 12.5 µm and 100 µm. b – i Representative images and quantification of Vimentin and MMP3 coimmunostaining ( b , f ), CD31 and EMCN coimmunostaining ( c , g ), Col2a1 immunofluorescence staining ( d , h ), and MMP13 immunohistochemical staining ( e , i ) in the knee joints of control and RA mice treated with vehicle or rmFABP4 for 4 and 8 weeks ( n = 10 per group). Scale bars: 12.5 µm, 25 µm, and 100 µm. One-way ANOVA and Tukey’s multiple comparison test. ** P < 0.01. The data are shown as the mean ± SEM

Article Snippet: We used human and mouse FABP4 ELISA kits (Elabscience Biotechnology, Bethesda, MD, USA: #E-EL-H0285c and #E-EL-M2404c) to analyze the level of FABP4 in the supernatant of macrophages stimulated with lipopolysaccharide (LPS), the serum of C57BL/6 J and TSC1KO mice, and human serum and synovial fluid.

Techniques: Recombinant, Immunohistochemical staining, Staining, Control, Immunofluorescence, Comparison

Journal: Bone Research

Article Title: FABP4 secreted by M1-polarized macrophages promotes synovitis and angiogenesis to exacerbate rheumatoid arthritis

doi: 10.1038/s41413-022-00211-2

Figure Lengend Snippet: Activation of the mTORC1 pathway enhances the secretion of FABP4 by M1-polarized macrophages to exacerbate RA progression. a , b Representative images and quantification of F4/80 and pS6 coimmunostaining in the mouse ( n = 10 per group) ( a ) and human synovium ( n = 16 per group) ( b ) of the control and RA groups. Scale bar: 12.5 µm. c Western blot showing S6 and pS6 in human synovial tissue. d Western blot showing FABP4, NOS2, S6 and pS6 in BMDMs treated with LPS or rapamycin. e Representative immunofluorescence staining images and quantitative analysis of FABP4 in BMDMs stimulated with LPS or rapamycin ( n = 3 per group). Scale bar: 100 µm. f Western blot showing FABP4, NOS2, S6 and pS6 in BMDMs treated with MHY1485 or rapamycin. g Representative immunofluorescence staining images and quantitative analysis of FABP4 in BMDMs treated with MHY1485 or rapamycin ( n = 3 per group). Scale bar: 100 µm. h Western blot showing FABP4, NOS2, S6 and pS6 in BMDMs from control and TSC1KO mice. i Representative images and quantification of F4/80 and pS6 coimmunostaining in the synovium of wild-type and TSC1KO mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bar: 12.5 µm. j Representative images and quantification of FABP4 immunohistochemical staining and coimmunostaining of FABP4 with F4/80 or NOS2 in the synovium of control and TSC1KO mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bars: 12.5 µm and 100 µm. f–j Representative images ( f ) and quantification of Vimentin and MMP3 coimmunostaining ( g ), CD31 and EMCN coimmunostaining ( h ), Col2a1 immunofluorescence staining ( i ), and MMP13 immunohistochemical staining ( j ) in the knee joints of controls and TSC1KO mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bar: 12.5 µm, 25 µm, and 100 µm. Student’s t -test or one-way ANOVA and Tukey’s multiple comparison test. ** P < 0.01. The data are shown as the mean ± SEM

Article Snippet: We used human and mouse FABP4 ELISA kits (Elabscience Biotechnology, Bethesda, MD, USA: #E-EL-H0285c and #E-EL-M2404c) to analyze the level of FABP4 in the supernatant of macrophages stimulated with lipopolysaccharide (LPS), the serum of C57BL/6 J and TSC1KO mice, and human serum and synovial fluid.

Techniques: Activation Assay, Control, Western Blot, Immunofluorescence, Staining, Immunohistochemical staining, Comparison

Journal: Bone Research

Article Title: FABP4 secreted by M1-polarized macrophages promotes synovitis and angiogenesis to exacerbate rheumatoid arthritis

doi: 10.1038/s41413-022-00211-2

Figure Lengend Snippet: Inhibition of the mTORC1 pathway reduces FABP4 secretion by M1-polarized macrophages to attenuate RA progression. a Representative images and quantification of F4/80 and pS6 coimmunostaining in the synovium of controls and Rheb1KO mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bar: 12.5 µm. b – e Representative images ( b ) and quantitative analysis of FABP4 immunohistochemical staining ( c ) and coimmunostaining of FABP4 with F4/80 ( d ) or NOS2 ( e ) in the synovium of controls and Rheb1KO mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bars: 12.5 µm and 100 µm. f – j Representative images ( f ) and quantitative analysis of Vimentin and MMP3 coimmunostaining ( g ), CD31 and EMCN coimmunostaining ( h ), Col2a1 immunofluorescence staining ( i ), and MMP13 immunohistochemical staining ( j ) in the knee joints of controls and Rheb1KO mice at 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bar: 12.5 µm, 25 µm, and 100 µm. Student’s t -test. ** P < 0.01. The data are shown as the mean ± SEM

Article Snippet: We used human and mouse FABP4 ELISA kits (Elabscience Biotechnology, Bethesda, MD, USA: #E-EL-H0285c and #E-EL-M2404c) to analyze the level of FABP4 in the supernatant of macrophages stimulated with lipopolysaccharide (LPS), the serum of C57BL/6 J and TSC1KO mice, and human serum and synovial fluid.

Techniques: Inhibition, Immunohistochemical staining, Staining, Immunofluorescence

Journal: Bone Research

Article Title: FABP4 secreted by M1-polarized macrophages promotes synovitis and angiogenesis to exacerbate rheumatoid arthritis

doi: 10.1038/s41413-022-00211-2

Figure Lengend Snippet: BMS309403 and anagliptin reduce FABP4 expression in murine synovial macrophages. a – c Representative images and quantification of FABP4 immunohistochemical staining and coimmunostaining of FABP4 with F4/80 or NOS2 in the synovium of TSC1KO mice treated with vehicle, BMS309403 or anagliptin for 4 and 8 weeks after AIA modeling ( n = 10 per group). Scale bar: 12.5 µm and 100 µm. d – f Representative images and quantification of FABP4 immunohistochemical staining and coimmunostaining of FABP4 and F4/80 or NOS2 in the synovium of RA mice treated with vehicle or BMS309403 for 4 and 8 weeks ( n = 10 per group). Scale bar: 12.5 µm and 100 µm. Student’s t -test or one-way ANOVA and Tukey’s multiple comparison test. * P < 0.05, ** P < 0.01. The data are shown as the mean ± SEM

Article Snippet: We used human and mouse FABP4 ELISA kits (Elabscience Biotechnology, Bethesda, MD, USA: #E-EL-H0285c and #E-EL-M2404c) to analyze the level of FABP4 in the supernatant of macrophages stimulated with lipopolysaccharide (LPS), the serum of C57BL/6 J and TSC1KO mice, and human serum and synovial fluid.

Techniques: Expressing, Immunohistochemical staining, Staining, Comparison

Journal: Bone Research

Article Title: FABP4 secreted by M1-polarized macrophages promotes synovitis and angiogenesis to exacerbate rheumatoid arthritis

doi: 10.1038/s41413-022-00211-2

Figure Lengend Snippet: Inhibiting FABP4 prevents RA development. a – d Representative images of Vimentin and MMP3 coimmunostaining ( a ), CD31 and EMCN coimmunostaining ( b ), Col2a1 immunofluorescence staining ( c ), and immunohistochemical staining of MMP13 ( D ) in the knee joints of TSC1KO mice treated with vehicle, BMS309403 or anagliptin for 4 and 8 weeks after AIA modeling. Scale bar: 12.5 µm, 25 µm, and 100 µm. e – h Representative images of Vimentin and MMP3 coimmunostaining ( e ), CD31 and EMCN coimmunostaining ( f ), Col2a1 immunofluorescence staining ( g ), and immunohistochemical staining of MMP13 ( h ) in the knee joints of RA mice treated with vehicle or BMS309403 for 4 and 8 weeks. Scale bar: 12.5 µm, 25 µm, and 100 µm. i – l Quantification of Vimentin-MMP3 ( i ), CD31-EMCN ( j ), Col2a1 ( k ), and MMP13 ( l ) in TSC1KO mice treated with vehicle, BMS309403, or anagliptin for 4 and 8 weeks after AIA surgery ( n = 10 per group). m – p Quantification of Vimentin-MMP3 ( m ), CD31-EMCN ( n ), Col2a1 ( o ), and MMP13 ( p ) in RA mice treated with vehicle or BMS309403 for 4 and 8 weeks ( n = 10 per group). Student’s t -test or one-way ANOVA and Tukey’s multiple comparison test were used. ** P < 0.01. The data are shown as the mean ± SEM

Article Snippet: We used human and mouse FABP4 ELISA kits (Elabscience Biotechnology, Bethesda, MD, USA: #E-EL-H0285c and #E-EL-M2404c) to analyze the level of FABP4 in the supernatant of macrophages stimulated with lipopolysaccharide (LPS), the serum of C57BL/6 J and TSC1KO mice, and human serum and synovial fluid.

Techniques: Immunofluorescence, Staining, Immunohistochemical staining, Comparison

Journal: Journal of Inflammation Research

Article Title: Wheat-Grain Moxibustion Ameliorates Ulcerative Colitis: Suppressing Intestinal Inflammation, Modulating Gut Microbiota, and Restoring Mucosal Barrier Integrity

doi: 10.2147/JIR.S540574

Figure Lengend Snippet: Wgm improves the intestinal mucosal barrier integrity in UC mice. ( A – D ) Serum levels of DAO, D-LA, ICAM-1, and IFABP (n=10). ( E – L ) Immunohistochemical analysis of Claudin 1, MUC2, Occludin, and ZO-1 in colonic tissues (n=4). Scale bars: 100 μm; 50 μm. ( M – P ) Western blot analysis of Claudin 1, Occludin, and ZO-1 expression in colonic tissues (n=3). Data are expressed as mean ± SEM. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Following the manufacturer’s protocols of ELISA kits, serum levels of ICAM-1 (KE10129, Proteintech), IFABP (E-EL-M0735, Elabscience), DAO (E-EL-M0412, Elabscience), and D-LA (E-BC-K002-M, Elabscience) were measured.

Techniques: Immunohistochemical staining, Western Blot, Expressing

Journal: iScience

Article Title: The mesenteric adipokine SFRP5 alleviated intestinal epithelial apoptosis improving barrier dysfunction in Crohn's disease

doi: 10.1016/j.isci.2024.111517

Figure Lengend Snippet: The adipokine SFRP5 was increased in the htMAT of CD patients (A) Representative image of IHC staining showing that SFRP5 expression was elevated in htMAT but was only visible in NL-MAT and nMAT. (B) Quantitative analysis of the results of the IHC staining for SFRP5. (C and D) The level of the SFRP5 protein in MAT was detected by WB. (E) The RT-qPCR result of SFRP5 in MAT. (F) The levels of SFRP5 in the serum of humans were determined by enzyme-linked immunosorbent assay (ELISA). CD, Crohn disease; NL, colon cancer; HC, healthy control; MAT, mesenteric adipose tissue; htMAT, hypertrophic MAT in patients with CD; nMAT, normal MAT in patients with CD; NL-MAT, normal MAT in patients with colon cancer. N = 18 per group. The data are presented as the mean ± SD. ANOVA (Tukey’s multiple test) was used for comparison of measurement data. ∗ p < 0.05. NS, not significant.

Article Snippet: ELISA kit (I-FABP) , BOSTER , EK1622.

Techniques: Immunohistochemistry, Expressing, Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Control, Comparison

Journal: iScience

Article Title: The mesenteric adipokine SFRP5 alleviated intestinal epithelial apoptosis improving barrier dysfunction in Crohn's disease

doi: 10.1016/j.isci.2024.111517

Figure Lengend Snippet: The protective effect of SFRP5 on CD-like colitis in vivo may be at least partially mediated by Wnt5a/JNK signaling (A–D) The levels of Wnt5a, p-JNK, and Bcl2 in the colonic mucosa were analyzed by WB. (E and F) The levels of TNF-α and IL-6 were detected via ELISA. (G) DAI scores. (H and I) Representative images of H&E staining of the whole colon and colonic inflammation scores. (J and K) FD4 levels in the serum and colonic TEER values. (L–P) The ZO-1, Claudin-1, and C-caspase3 protein levels were detected by WB. WT, wild-type; TNBS, 2,4,6-trinitrobenzenesulfonic acid; SFRP5, rAAV9-SFRP5; DAI, disease activity index; FD4, fluorescein isothiocyanate-dextran (4 kDa); TEER, transepithelial electrical resistance; ZO-1, zonula occludens-1; C-caspase3, cleaved caspase3. N = 6 per group. The data are presented as the mean ± SD. ANOVA (Tukey’s multiple test) was used for comparison of measurement data. ∗ p < 0.05.

Article Snippet: ELISA kit (I-FABP) , BOSTER , EK1622.

Techniques: In Vivo, Enzyme-linked Immunosorbent Assay, Staining, Activity Assay, Comparison

Journal: iScience

Article Title: The mesenteric adipokine SFRP5 alleviated intestinal epithelial apoptosis improving barrier dysfunction in Crohn's disease

doi: 10.1016/j.isci.2024.111517

Figure Lengend Snippet:

Article Snippet: ELISA kit (I-FABP) , BOSTER , EK1622.

Techniques: Recombinant, SYBR Green Assay, Enzyme-linked Immunosorbent Assay, In Situ, Software