Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: Irisin inhibited the formation of NETs in STZ-induced T2DM mouse model and improved T2DM. A . Glucose tolerance test evaluated the glucose tolerance of T2DM mice treated with Irisin, N = 5; B . ELISA detected the plasma insulin levels of T2DM mice treated with Irisin, N = 5; C - D . ELISA detected the levels of Cit-H3 and MPO in the plasma of T2DM mice treated with Irisin, N = 5; E . ELISA detected the serum Irisin levels of T2DM mice treated with Irisin, N = 3; F - G . Western blotting detected the expression levels of related proteins in pyroptosis and phosphorylation levels of STING and IRE1a, N = 3; H . HE staining detected the morphology of pancreatic tissue in T2DM mice treated with Irisin, N = 3; I . Immunofluorescence staining detected the levels of Cit-H3 and MPO in pancreatic tissue of T2DM mice treated with Irisin, N = 3; ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Enzyme-linked Immunosorbent Assay, Clinical Proteomics, Western Blot, Expressing, Phospho-proteomics, Staining, Immunofluorescence

Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: NETs promoted high glucose-induced Min6 cell pyroptosis, but Irisin could inhibit NETs. A . ELISA detected insulin levels in Min6 culture media under different treatment conditions; B - D . Western blotting detected the expression levels of GSDMD-N and C-caspase-1 in Min6 cells under different treatment conditions; E - F . Flow cytometry detected Min6 cell pyroptosis under different treatment conditions; G . Sytox green staining marked the levels of NETs released under different conditions; N = 3, * P < 0.05, ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Flow Cytometry, Staining

Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: Pyroptosis induced by NETs was regulated by NLRP1. A . Western blotting was performed to detect the expression of related proteins in pyroptosis in Min6 cells with NLRP1 knocked down; B . ELISA was used to detect the LDH, IL-1βand IL-18; C - D . Flow cytometry was used to detect pyroptosis in NLRP1 knocked down Min6 cells induced by NETs; N = 3, * P < 0.05, ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Western Blot, Expressing, Enzyme-linked Immunosorbent Assay, Flow Cytometry

Journal: Diabetology & Metabolic Syndrome

Article Title: Irisin regulates integrin αvβ5/FAK/ERK to inhibit neutrophil extracellular traps formation and reduce pancreatic beta-cells pyroptosis in type 2 diabetes mellitus

doi: 10.1186/s13098-025-01852-z

Figure Lengend Snippet: Pyroptosis induced by NETs was regulated by STING/IRE1α. A . ELISA was used to detect the LDH, IL-1β and IL-18; B - C . Western blotting was performed to detect the expression of related proteins in pyroptosis in Min6 cells with C-176 added; D - E . Flow cytometry was used to detect pyroptosis in Min6 cells with C-176 added induced by NETs; N = 3, * P < 0.05, ** P < 0.01

Article Snippet: Mouse Irisin ELISA kit (E-EL-M2743), LDH ELISA kit (E-EL-M0419), IL-18 ELISA kit (E-EL-M0730) were purchased from Elabscience.

Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing, Flow Cytometry

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Bradykinin/bradykinin 1 receptor promotes brain microvascular endothelial cell permeability and proinflammatory cytokine release by downregulating Wnt3a

doi: 10.1002/jbt.23213

Figure Lengend Snippet: BK enhanced ROS production, cellular permeability, and inflammation, and reduced cell proliferation and tight junction formation by BMECs. BMECs were treated with BK (low concentration, 100 nM or high concentration, 400 nM). ROS production was determined by flow cytometry, and the respective images are shown (A, left panel). Summarized results from three independent experiments are shown (A, right panel). Cell permeability was tested by sodium fluorescein analysis (B, left panel) and the lucifer yellow assay (B, right panel). Cell proliferation was measured by EdU staining. (C, IL‐6, MCP‐1, and IL‐18 expressions were detected by IF (D) and ELISA (E). (F) The cell apoptosis rate was measured using FCM. (G) LDH levels were determined by ELISA. The levels of claudin‐5, occludin, β‐catenin, Wnt3a, and B1R expression were determined by western blotting (H, left panel). The statistical results of western blot studies conducted in three independent experiments (H, right panel). * p < 0.05; ** p < 0.01; *** p < 0.001 versus blank. B1R, bradykinin 1 receptor; BK, bradykinin; BMECs, brain microvascular endothelial cells; DCF‐A, 2′,7′‐dichlorodihydrofluorescein diacetate; EdU, 5‐ethynyl‐2′‐deoxyuridine; ELISA, enzyme‐linked Immunosorbent assay; FCM, flow cytometry; GAPDH glyceraldehyde‐3‐phosphate dehydrogenase; IF, immunofluorescence; IL‐6, interleukin 6; LDH, lactate dehydrogenase; MCP‐1, monocyte chemoattractant protein‐1; ROS, reactive oxygen species.

Article Snippet: The levels of IL‐6 (Elabscience; Cat. No. E‐EL‐H0102c), IL‐18 (Boster; CAT. No. EK0864), MCP‐1 (Boster; Cat. No. EK0441), and LDH (Elabscience; CAT. No. E‐EL‐H0866c) expression were determined by enzyme‐linked immunosorbent assay (ELISA) according to manufacturer's instructions.

Techniques: Permeability, Concentration Assay, Flow Cytometry, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Immunofluorescence

Journal: Journal of Biochemical and Molecular Toxicology

Article Title: Bradykinin/bradykinin 1 receptor promotes brain microvascular endothelial cell permeability and proinflammatory cytokine release by downregulating Wnt3a

doi: 10.1002/jbt.23213

Figure Lengend Snippet: Wnt3a counteracted the effects of BK on BMECs. BMECs were treated with a high concentration (400 nM) of BK or a high concentration of both BK and Wnt3a (100 ng/ml each). ROS production was measured by flow cytometry, and the respective images are shown (A, left panel). The summary results of three independent experiments are shown (A, right panel). Cell permeability was tested by sodium fluorescein analysis (B, left panel) and the lucifer yellow assay (B, right panel). (C) Cell proliferation was measured by EdU staining. IL‐6, MCP‐1, and IL‐18 expressions were detected by (D) IF and (E) ELISA. (F) Cell apoptosis was detected by using the FCM method. (G) LDH levels were determined by ELISA. The levels of claudin‐5, occludin, β‐catenin, Wnt3a, and B1R expression were determined by western blotting (H, left panel). The statistical results of western blot studies conducted in three independent experiments (H, right panel). * p < 0.05; ** p < 0.01; *** p < 0.001 versus blank. # p < 0.05; ## p < 0.01 versus BK‐high. B1R, bradykinin 1 receptor; BK, bradykinin; BMECs, brain microvascular endothelial cells; DCF‐A, 2′,7′‐dichlorodihydrofluorescein diacetate; EdU, 5‐ethynyl‐2′‐deoxyuridine; ELISA, enzyme‐linked Immunosorbent assay; FCM, flow cytometry; GAPDH glyceraldehyde‐3‐phosphate dehydrogenase; IF, immunofluorescence; IL‐6, interleukin 6; LDH, lactate dehydrogenase; MCP‐1, monocyte chemoattractant protein‐1; ROS, reactive oxygen species.

Article Snippet: The levels of IL‐6 (Elabscience; Cat. No. E‐EL‐H0102c), IL‐18 (Boster; CAT. No. EK0864), MCP‐1 (Boster; Cat. No. EK0441), and LDH (Elabscience; CAT. No. E‐EL‐H0866c) expression were determined by enzyme‐linked immunosorbent assay (ELISA) according to manufacturer's instructions.

Techniques: Concentration Assay, Flow Cytometry, Permeability, Staining, Enzyme-linked Immunosorbent Assay, Expressing, Western Blot, Immunofluorescence

Journal: Arthritis Research & Therapy

Article Title: In vitro and in vivo assessment of the proresolutive and antiresorptive actions of resolvin D1: relevance to arthritis

doi: 10.1186/s13075-019-1852-8

Figure Lengend Snippet: RvD1 does not impair RAW264.7 cell viability. RAW264.7 cells were incubated with different concentrations of RvD1 (0–500 nM) without or with 50 ng/ml LPS for 72 h. Cell viability percentages were obtained by 3-(4,5-dimethyl-thiazoyl)-2,5-diphenyl-SH-tetrazolium bromide assay ( a ), and LDH release was measured in cell supernatants by a commercial kit ( b ). Cell death and LDH release were induced by the addition of 0.5 mM H 2 O 2 . Beclin-1 protein expression was assessed in cell extracts by western blot ( c ). ANOVA test was performed to compare each condition. Results are expressed as mean ± SEM for n = 4. Data are means ± SEM, and one-way ANOVA was performed to compare the results. ## p < 0.01 compared to untreated cells

Article Snippet: RvD1, LDH ELISA kit, and PGE 2 EIA kit were obtained from Cayman Chemical (Ann Arbor, Ml, USA).

Techniques: Incubation, Expressing, Western Blot

Journal: Arthritis Research & Therapy

Article Title: In vitro and in vivo assessment of the proresolutive and antiresorptive actions of resolvin D1: relevance to arthritis

doi: 10.1186/s13075-019-1852-8

Figure Lengend Snippet: RvD1 inhibits osteoclast activation and recruitment as well as LPS-induced TRAP, cathepsin k, PGE 2 , TNF-α, and RANK expression and concurrently enhances IL-10 release in LPS-activated RAW 264.7 macrophages. Macrophages were stimulated with LPS (50 ng/ml) with or without RvD1 treatment (0–500 nM) for 72 h. Cell extracts were collected, and then TRAP and cathepsin k protein expression was assessed by western blot ( a ). The same treatment was conducted after incubation or not of cells with sc-siRNA or siRNA against FPR2 (50 nM) for 24 h. TRAP enzymatic staining was performed, and TRAP-positive cells were observed with an inverted microscope (× 200). Arrows show osteoclasts with three nuclei or more ( b ). Western blot was employed to confirm FPR2 silencing by siRNA ( b ). In parallel, cell media was collected and then PGE 2 ( c ), TNF-α ( d ), IL-10 ( e ), and RANK ( f ) expression was assessed by commercial kits. Data are means ± SEM for n = 3, and one-way ANOVA was performed to compare the results. # p < 0.05, ## p < 0.01, ### p < 0.001, and #### p < 0.0001 compared to non-stimulated cells; * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001 compared to LPS-activated cells

Article Snippet: RvD1, LDH ELISA kit, and PGE 2 EIA kit were obtained from Cayman Chemical (Ann Arbor, Ml, USA).

Techniques: Activation Assay, Expressing, Western Blot, Incubation, Staining, Inverted Microscopy

Journal: Arthritis Research & Therapy

Article Title: In vitro and in vivo assessment of the proresolutive and antiresorptive actions of resolvin D1: relevance to arthritis

doi: 10.1186/s13075-019-1852-8

Figure Lengend Snippet: RvD1 inhibits hydroxyapatite matrix degradation as well as bone resorption induced by primary human monocyte-derived osteoclasts. a Human monocytes were incubated on a hydroxyapatite matrix. They were first treated without or with sc-siRNA or siRNA FPR2 (50 nM) for 24 h and then stimulated over 2 weeks with RANKL (50 ng/ml) and M-CSF (10 ng/ml) with or without RvD1 treatment (500 nM). Culture medium was changed every 3 days. Von Kossa staining was performed, then pit areas were observed under an inverted microscope (× 200) and measured using ImageJ software. b Normal mouse femoral bone explants were incubated with RANKL (50 ng/ml) and M-CSF (10 ng/ml) with or without RvD1 treatment (100 or 500 nM) over 28 days. Culture medium was changed every 3 days. Hematoxylin-eosin staining was performed, and resorption areas were observed and scored under an inverted microscope (× 200). Data are means ± SEM for n = 3, and one-way ANOVA was performed to compare the results. ## p < 0.01 compared to untreated cells; * p < 0.05 compared to M-CSF/RANKL-activated cells; @ p < 0.05 compared to RvD1-stimulated cells

Article Snippet: RvD1, LDH ELISA kit, and PGE 2 EIA kit were obtained from Cayman Chemical (Ann Arbor, Ml, USA).

Techniques: Derivative Assay, Incubation, Staining, Inverted Microscopy, Software

Journal: Arthritis Research & Therapy

Article Title: In vitro and in vivo assessment of the proresolutive and antiresorptive actions of resolvin D1: relevance to arthritis

doi: 10.1186/s13075-019-1852-8

Figure Lengend Snippet: RvD1 treatment improves RA clinical endpoints as well as synovium and bone changes in arthritic mice. Arthritis was induced by i.p. injection of arthritogenic cocktail of five monoclonal anti-type II collagen monoclonal antibodies (mAb) on day 1, followed by i.p. injection of 50 μg LPS on day 4, as described in the “ ” section. Over the course of the experiment, and for each of the experimental groups (non-immunized control mice, CAIA mice immunized with type II collagen antibody and treated with vehicle, and mice immunized with type II collagen and treated with RvD1 (100, 500, 1000 ng/day) at day 1 or with 1000 ng/ml at day 4, clinical score was attributed by two investigators blinded to groups on a scale of 0–4 for each paw, for a maximum of 16 for all four paws, as described in the “ ” section ( a ). Hind paw thickness was measured using a caliper and expressed as thickness change from baseline ( b ). Weight evolution was assessed and expressed as percentage of weight loss from baseline ( c ). Data are means ± SEM for n = 6, and one-way ANOVA was performed to compare the results. * p < 0.05 and ** p < 0.01 compared to RA group. Cell proliferation in synovial membrane and TRAP staining in bone were performed as described in the “ ” section ( d )

Article Snippet: RvD1, LDH ELISA kit, and PGE 2 EIA kit were obtained from Cayman Chemical (Ann Arbor, Ml, USA).

Techniques: Injection, Bioprocessing, Control, Membrane, Staining

Journal: Arthritis Research & Therapy

Article Title: In vitro and in vivo assessment of the proresolutive and antiresorptive actions of resolvin D1: relevance to arthritis

doi: 10.1186/s13075-019-1852-8

Figure Lengend Snippet: RvD1 daily treatment prevents cartilage and bone alterations in arthritic mouse joints. Arthritis was induced as previously described. After mouse sacrifice, blood was collected. Knees and ankles were removed and then fixed and sectioned as described in the “ ” section. Knee sections were stained with safranin-O staining and scored ( a ). Ankles were examined with a micro-CT scanner ( b ). Serum RvD1, TRAP, CTX-I, and CTX-II levels were assessed by EIA and ELISA, respectively ( c – f ). Data are means ± SEM, and one-way ANOVA was performed to compare the results. # p < 0.05, ## p < 0.01, and #### p < 0.0001 compared to non-immunized group; * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to CAIA group. n = 4–5 mice per group. F femur, T tibia

Article Snippet: RvD1, LDH ELISA kit, and PGE 2 EIA kit were obtained from Cayman Chemical (Ann Arbor, Ml, USA).

Techniques: Staining, Micro-CT, Enzyme-linked Immunosorbent Assay

Journal: Arthritis Research & Therapy

Article Title: In vitro and in vivo assessment of the proresolutive and antiresorptive actions of resolvin D1: relevance to arthritis

doi: 10.1186/s13075-019-1852-8

Figure Lengend Snippet: RvD1 daily treatment decreases inflammatory mediators’ release in arthritic mouse serum. Arthritis was induced as previously described in Fig. . Blood was collected after mice sacrifice at day 10. Serum levels of inflammatory cytokines ( a – e ) as well as PGE 2 ( f ) were measured by multiplex and EIA assays respectively. Data are means ± SEM. One-way ANOVA was performed to compare the results. * p < 0.05, ** p < 0.01, and *** p < 0.001 compared to CAIA group; # p < 0.05, ## p < 0.01, ### p < 0.001, and #### p < 0.0001 compared to non-immunized group. n = 4–5 mice per group

Article Snippet: RvD1, LDH ELISA kit, and PGE 2 EIA kit were obtained from Cayman Chemical (Ann Arbor, Ml, USA).

Techniques: Multiplex Assay

Journal: BMC Biotechnology

Article Title: Method for quantification of porcine type I interferon activity using luminescence, by direct and indirect means

doi: 10.1186/s12896-022-00743-9

Figure Lengend Snippet: Activities of SGLuc-Δ1D2A-IFNα and IFNα-Δ1D2A-SGLuc Δ1M

Article Snippet: Concentrations of porcine IFNα and SGLuc-IFNα samples were determined using a Porcine IFNα ELISA kit (Millipore Sigma, RAB1131-1KT), and a Porcine Interferon Beta ELISA kit (Novateinbio, NB-E50024) was used for IFNβ and SGLuc-IFNβ samples.

Techniques: Concentration Assay, Enzyme-linked Immunosorbent Assay