Journal: Kidney Diseases

Article Title: Treatment of Membranous Nephropathy by Disulfiram through Inhibition of Podocyte Pyroptosis

doi: 10.1159/000524164

Figure Lengend Snippet: DSF alleviated C3a/C5a-induced podocyte injury by inhibiting pyroptosis. Podocyte injury was induced with C3a (50 nM) and C5a (50 nM). The inhibitory effects of DSF (250 nM) on C3a/C5a-induced pyroptosis were examined. a Representative images of GSDMD(N)/ZO-1/Nucleus (DAPI) triple immunofluorescent staining of treated podocytes. Scale bars = 20 μm. b Representative Western Blot images of GSDMD, NF-κB p65, p-NF-κB p65 (Ser536), NLRP3, ASC, Caspase-1, IL-18 and the internal control (GAPDH) in treated podocytes. c IL-18 release in treated podocytes was detected. d, e Representative images of PI/Nucleus (DAPI) double fluorescent staining of treated podocytes ( d ) and percentage of PI-positive cells ( e ); arrows, PI-positive cells; scale bars = 40 μm. f LDH release in treated podocytes was detected. The data above represent three independent experiments in duplicate and are shown as the mean ± SD, and ANOVA with LSD-t test (equal variances assumed) or Welch's test with Dunnett's T3 test (equal variances not assumed) was used for multiple comparisons among groups. *, p < 0.05; **, p < 0.01.

Article Snippet: IL-18 release in the culture supernatant of the podocytes and the serum IL-1β/IL-18 levels of rats were tested using human IL-18 ELISA and rat IL-1β/IL-18 ELISA kits (Boster Biological Technology Co. Ltd., Wuhan, China), respectively.

Techniques: Staining, Western Blot

Journal: Kidney Diseases

Article Title: Treatment of Membranous Nephropathy by Disulfiram through Inhibition of Podocyte Pyroptosis

doi: 10.1159/000524164

Figure Lengend Snippet: DSF inhibited the renal pyroptosis signaling pathway in PHN rats. a Representative renal GSDMD(N)/Synaptopodin and GSDMD(N)/ZO-1 double immunofluorescent staining of the rats ( n = 6) in each group; Scale bars = 20 μm. b, c Representative renal immunohistochemical staining ( b ) and semiquantification based on the glomerular IOD/area of GSDMD(N), NF-κB p65, p-NF-κB p65 (Ser536), NLRP3, ASC, Caspase-1, IL-1β, and IL-18 ( c ) of the rats ( n = 6) in each group; Scale bars = 20 μm. d Relative mRNA levels of glomerular GSDMD, NLRP3, ASC, Caspase-1, IL-1β and IL-18 of the rats ( n = 6) in each group. e Representative Western Blot images of renal GSDMD, NF-κB p65, p-NF-κB p65 (Ser536), nuclear NF-κB p65, nuclear p-NF-κB p65 (Ser536), NLRP3, ASC, Caspase-1, Caspase-1 p20, IL-1β, IL-1β (mature form), IL-18 and the internal control (GAPDH, Histone H3) of the rats ( n = 6) in each group. Serum IL-1β ( f ) and IL-18 ( g ) of the rats ( n = 6) in each group on days 1, 5, 8, 15 after model establishment. The data above are shown as the mean ± SD ( c, d, f, g ) and were compared to the PHN group ( f, g ). ANOVA with LSD-t test (equal variances assumed) or Welch's test with Dunnett's T3 test (equal variances not assumed) was used for multiple comparisons among groups. *, p < 0.05; **, p < 0.01.

Article Snippet: IL-18 release in the culture supernatant of the podocytes and the serum IL-1β/IL-18 levels of rats were tested using human IL-18 ELISA and rat IL-1β/IL-18 ELISA kits (Boster Biological Technology Co. Ltd., Wuhan, China), respectively.

Techniques: Staining, Immunohistochemical staining, Western Blot

Journal: Oncology reports

Article Title: Protective effect of neutralizing anti-IL-18α monoclonal antibody on a mouse model of acute graft-versus-host disease.

doi: 10.3892/or.2015.4176

Figure Lengend Snippet: Figure 1. Anti-IL-18Rα mAb alleviates aGVHD systemic symptoms in aGVHD mice. aGVHD was induced in irradiated BALB/c mice by BMC transplanta- tion. Ten micrograms of anti-IL-18Rα mAb was administered by intraperitoneal injection to mice in the BS+Ab group, while the BS group received injection of PBS. (A) Hunch posture, hair loss and skin lesions were evaluated on day 14 P.T. (B) Changes in body weight of the recipient mice at different time‑points in the different groups. (C) aGVHD clinical scores of the BS+Ab and BS groups on day 14 P.T. (D) Survival rates of aGVHD mice in the different groups. n=6 in each group. *P<0.05. GVHD, graft-versus-host disease; IL-18, interleukin-18; aGVHD, acute GVHD; BMC, bone marrow cell; PBS, phosphate-buffered solution; mAb, monoclonal antibody; P.T., post‑transplantation.

Article Snippet: Recipient mice in the BS+Ab group also received 10 μg/mouse intraperitoneal injection of neutralizing mAb against murine IL-18Rα (catalog no. MAB12161; R&D Systems, Minneapolis, MN, uSA) every 2 days.

Techniques: Irradiation, Injection

Journal: Oncology reports

Article Title: Protective effect of neutralizing anti-IL-18α monoclonal antibody on a mouse model of acute graft-versus-host disease.

doi: 10.3892/or.2015.4176

Figure Lengend Snippet: Figure 2. Effect of anti-IL-18Rα mAb administration on Th cell subsets, pro-inflammatory cytokines and histological scores in the aGVHD mice. (A-C) Peripheral blood levels of Th1 (A), Th2 (B) and Th17 (C) cell subsets in the BS+Ab and BS experimental groups were measured by flow cytometry at different time-points. Serum levels of IFN-γ (D), IL-4 (E), IL-17A (F) and IL-6 (H) at different time-points were detected by cytometric bead array, and IL-18 levels (G) were measured by ELISA. (I) Representative H&E staining of the liver and small intestine tissues of the mice in the BS+Ab and BS groups and the normal control group (untreated animals) on day 14 P.T. Magnification, x400. (J) Histological score was measured on day 14 P.T. n=6 in each group, *P<0.05. GVHD, graft-versus-host disease; aGVHD, acute GVHD; mAb, monoclonal antibody; Th, T helper; IL-18, interleukin-18; ELISA, enzyme-linked immunosorbent assay; H&E, hematoxylin and eosin; P.T., post‑transplantation.

Article Snippet: Recipient mice in the BS+Ab group also received 10 μg/mouse intraperitoneal injection of neutralizing mAb against murine IL-18Rα (catalog no. MAB12161; R&D Systems, Minneapolis, MN, uSA) every 2 days.

Techniques: Flow Cytometry, Enzyme-linked Immunosorbent Assay, Staining, Control