Fig 2: TSP1 promoted the differentiation of cmMφ, and enhanced migration. (A-C) In vitro, TSP1 protein promoted the differentiation of LPMC to CD11c+ LPMP and inhibited the cell proliferation of LPMC and CD11c+ LPMP (IF, 200×). (D) The number of CD11c+ LPMP in LPMC treated with MLS. (E) In vitro, TSP1 protein inhibited the cell proliferation of LPMC and CD11c+ LPMP. (F) The migration of LPMC and CD11c+ LPMP induced by IL-1β. MLS, mucosal lysis supernatant; LPMC, lamina propria mononuclear cells. (*, P<0.05, **, P<0.001, #, P<0.05, ##, P<0.001, vs. Con). MLS, mucosal lysis supernatant; IL, interleukin; anti-TSP1, anti-TSP1 antibody; Con, Control group; Mod, Model group; LSKL, an inhibitor of Thrombospondin (TSP-1); LPMC, Lamina propria mononuclear cells; LPMP, Lamina propria macrophages; Mφ, macrophage.

Fig 3: Increased thomsbospondin-1 (TSP1) expression in mice after IH exposure.(A) C57BL/6 mice were housed under normoxia or IH with or without infusion of Ang II for 28 days. Plasma concentration of TSP1 in mice detected by ELISA. (B) Thbs1 mRNA level in left ventricle of mice exposed to IH with or without infusion of Ang II for 14 days quantified by RT-PCR. (C) C57BL/6 mice were housed under normoxia or IH with or without infusion of Ang II for 28 days. The protein levels of TSP1, phosphorylated Smad2/3 (p-Smad2/3), and total Smad2/3 (t-Smad2/3) in left ventricle of mice detected by western blot analysis. (D) Quantification of TSP1 and p-Smad2/3 in (C). (E) Representative confocal microscopy images of immunofluorescence staining for TSP1 and DAPI. Scale bar, 20 μm. (F) Quantification of TSP1 fluorescent intensity in (E). Data are mean ± SEM, n = 6 mice per group, *p<0.05, 2-way ANOVA with Bonferroni post-test. (G) Quantification of number of cells negative for CD45, CD31, CD11b, and Ter119 and positive for Thy1 from left ventricle tissue digestion, that stained positive for intracellular TSP1 by flow cytometry. Data are mean ± SEM, n = 6, *p<0.05, unpaired 2-tail t test. (H) Plasma concentration of TSP1 in healthy individuals and patients with obstructive sleep apnea (OSA) detected by ELISA. Data are mean ± SEM, n = 21, *p<0.05, unpaired 2-tail t test. Data used for quantitative analyses as well as the numerical data that are represented in graphs are available in Figure 2—source data 1.Figure 2—source data 1.Intermittent hypoxia (IH) increased thomsbospondin-1 (TSP1) expression.

Fig 4: Downregulation of miR-205 expression in hypertrophic scars. (A) Different expression of miRNAs in adjacent skin and hypertrophic scars was accessed by miRNA microarray analysis. (B, C) miR-205 expression in hypertrophic scars tissues and fibroblasts was examined. **P < 0.05. (D) The schematic of binding sites between miR-205 and THBS1 3’-UTR. (E, F) THBS1 expression in hypertrophic scars tissues and fibroblasts was examined. **P < 0.05. (G) The correlation between miR-205 and THBS1 expression was estimated. P < 0.05 was considered statistically significant. (H) The relative dual-luciferase activity of THBS1 in cells expressing miR-205 was determined by dual-luciferase assay. **P < 0.05, vs. control.

Fig 5: The PRKCQ/NF-κB signaling pathway played an important role in regulating the effect of TSP1 protein. (A-C) The expression of CD36, PRKCQ, phospho-PRKCQ (Ser-676), phospho-PRKCQ (Thr-538), phospho-PRKCQ (Ser-695), NF-κB, and phospho-65 NF-κB in LPMC and CD11c+ LPMP. (D-F) The expression of CD36, PRKCQ, phospho-PRKCQ (Ser-676), phospho-PRKCQ (Thr-538), phospho-PRKCQ (Ser-695), NF-κB, and phospho-65 NF-κB in group stimulated with CD36over plasmid, CD36− plasmid, and TSP1 protein. (**, P<0.001, #, P<0.05, ##, P<0.001, ^, P<0.05, ^^, P<0.001, vs. Con). Con, Control group; TSP1, group treated with TSP1 protein; LPMC, Lamina propria mononuclear cells; LPMP, Lamina propria macrophages; CD36over, CD36-overexpression plasmid; CD36−, CD36 silencing plasmid.