Fig 1: Neutrophil infiltration depends on ICAM-1 in the cochlea, and ICAM-1 expression is lower in the stria vascularis than in the spiral ligament. The in vivo blockade of the ICAM-1 ligands (LFA-1 and Mac-1) decreases neutrophil infiltration in the cochlea after LPS inoculation. (A) Flow cytometry analysis showed a statistically significant difference in the number of neutrophils (Ly6G+, CD11b+) in the cochlea with LPS without blocking and LPS with anti-Mac-1 antibody treatment in vivo. (B) The fold change in mRNA expression at 1 day after LPS injection compared to basal conditions analyzed by qPCR. Notably, ICAM-1 mRNA expression was not increased in the stria vascularis, although IL-1ß mRNA expression was increased. (C) Western blot analysis using anti-ICAM-1 antibody. Beta-actin was blotted on the same membrane after stripping. An representative image of Western blot was shown from three independent experiments. Six cochleae from three mice were pooled for each group per experiment. (D) Western blot was quantified by the intensity of ICAM-1 band normalized by the beta-actin band. Relative expression was compared to 'SV Ctr' (untreated group) lane. SL, spiral ligament. SV, stria vascularis (** denotes P-value < 0.005, in post-hoc analysis after ANOVA).
Fig 2: IL-6 promotes VCAM-1, ICAM-1 and VEGF expression. (A) Reverse transcription-quantitative PCR and (B) ELISA demonstrated that IL-6 significantly promoted the expression of VCAM-1, ICAM-1, and VEGF in OLP-MFs and NFs. MRA significantly decreased the secretion of vascular-related growth factors in the OLP-MFs group compared with the control group. The expression of angiogenesis-associated cytokines in the OLP-MFs group were significantly higher compared with the NFs group. Control group, cells without treatment; IL-6 group, cells were treated with IL-6/sIL-6R; IL-6+MRA group, cells were treated with IL-6/sIL-6R and MRA. *P<0.05, **P<0.01 and ***P<0.001. IL-6, interleukin 6; VCAM-1, vascular cell adhesion molecule 1; ICAM-1, intercellular adhesion molecular 1; VEGF, vascular endothelial growth factor; OLP, oral lichen planus; MFs, myofibroblasts; NFs, normal fibroblasts; MRA, tocilizumab; ns, not significant.
Fig 3: Schematic figure of the working model diagram of the regulatory function of platelet-derived miR-223 in endothelial cells. In the case of KD, platelets are hyperactive and internalized into HCAECs. The horizontal transfer of platelet derives miR-223 to HCAECs, which suppress the expression of ICAM-1 in HCAECs and partially attenuates leukocyte adhesion, thereby reducing further endothelial damage in KD vasculitis. ICAM-1, intercellular cell adhesion molecule-1.
Fig 4: Validation of the mechanisms of GLXB treatment against AS in vivo. (A) The level of IL-6 in the serum of mice. (B) The level of IL-1ß in the serum of mice. (C) The level of TNF-a in the serum of mice. (D) The level of ALOX5 in the serum of mice. (E) The expression of the p-p38 protein in the aorta of mice. (F) The level of PTGS2 in the serum of mice. (G) The expression of the p-AKT protein in the aorta of mice. (H) The level of VEGFA in the serum of mice. (I) The level of eNOS in the serum of mice. Data were expressed as mean ± SEM. * p < 0.05, ** p < 0.01. (J) The expressions of VCAM-1 and ICAM-1 proteins in the aorta of mice. Data were expressed as mean ± SEM. ** p < 0.01 vs. control group, ?? p < 0.01 vs. GLXB group, # p < 0.05, ## p < 0.01 vs. model group. (K) Coimmunofluorescence staining of aortic tissue sections with antibodies to CD68 and a-SMA. Bar = 25 µm. (L) The level of TC in the serum of mice. (M) The level of TG in the serum of mice. (N) The level of LDL-C in the serum of mice. (O) The level of HDL-C in the serum of mice. Data were expressed as mean ± SEM (n = 6). * p < 0.05, ** p < 0.01. GLXB: Gualou–Xiebai herb pair (6 g/kg); GL: Gualou (4 g/kg); XB: Xiebai (2 g/kg); ATO: atorvastatin (10 mg/kg).
Fig 5: In LCWE-induced KD murine model, deficiency of platelet-miR-223 exacerbated the medial thickening, increased ICAM-1 expression with concomitant CD45+ inflammatory cells infiltration in the endothelium of abdominal aorta. LCWE or PBS was administrated i.p. for two weeks. The abdominal aorta tissues were collected, and continuous cross-sections were performed for H&E staining and immunofluorescence analysis. (A) Representative H & E-stained sections of PBS or LCWE injected mice were shown. n = 4, Scale bar: 50μm. (B) Quantification of the media layer areas of abdominal aorta in each group was shown. n = 4, Data are presented as mean ± SD, One-way ANOVA and Tukey’s multiple comparisons test. (C) Representative immunofluorescence images of sections from PBS or LCWE-injected mice. CD31 was stained as red, ICAM-1 as green, and the nucleus visualized as blue (DAPI). n = 4, Scale bars: 20μm. (D) Quantification of ICAM-1 expression in abdominal aorta tissues was shown (n = 4). Data are presented as mean ± SD, One-way ANOVA and Tukey’s multiple comparisons test. (E) Representative immunofluorescence images of sections from PBS or LCWE-injected mice. CD31 was stained as red, CD45 as green, and the nucleus visualized as blue (DAPI). n = 4, Scale bars: 20μm. (F) Quantification of CD45 expression in abdominal aorta tissues was shown (n = 4). Data are presented as mean ± SD, One-way ANOVA and Tukey’s multiple comparisons test. L, lumen; A, adventitia; Flox ctrl: miR-223 flox/flox mice.PF4-miR-223 KO, PF4-cre: miR-223 flox/flox mice. * P < 0.05, *** P < 0.001, **** P < 0.0001.
Supplier Page from Abcam for Anti-ICAM1 antibody [EPR22161-284]