Table 1.
The primer sequences for relative mRNA used in this study.
Fig 1.
THY1 promotes dextran sulfate sodium (DSS)-induced colitis in mice.
(A, B) The knockdown and overexpression of THY1 expression levels in NCM460 cells and DSS-induced colitis mouse colon tissue samples were determined by quantitative real-time PCR and western blot analysis, respectively. (C) Body weight loss and (D) Appearance of colon length of DSS-induced colitis mouse in different treatment groups including the Sham (blank control) group, the IBD model group, the IBD + OE-THY1 group, the IBD + OE-NC group. the IBD + KD-THY1 group, and the IBD + sh-NC group. (E) Representative H&E staining images of DSS-induced colitis mouse colon samples in different treatment groups. The corresponding image below is a high-magnification view of the red square position in the image above. A minimum of three separate experiments were carried out and the data presented are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 2.
THY1 promotes the pathological process of DSS-induced colitis.
(A, B) The expression levels of inflammatory factors (IL-1β, TNF-α and TGF-β), oxidative stress-related molecules (ROS, MDA and GSH) and angiogenesis-related stimulatory factors (HIF-1α and VEGF) in mouse colon tissue samples and NCM460 cell homogenate of co-culture system were detected by enzyme-linked immunosorbent assay (ELISA). A minimum of three separate experiments were carried out and the data presented are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 3.
THY1 inhibits M2 macrophage polarization.
(A, B) Macrophage immunophenotyping (M1 polarization and M2 polarization) were evaluated by flow cytometry in co-culture system of control group (Upper layer: THP-1 cells; Lower layer: normal NCM460 cells), sh-THY1 group (Upper layer: THP-1 cells; Lower layer: sh-THY1 treated NCM460 cells), sh-NC group (Upper layer: THP-1 cells; Lower layer: sh-NC treated NCM460 cells), OE-THY1 group (Upper layer: THP-1 cells; Lower layer: OE-THY1 treated NCM460 cells), and OE-NC group (Upper layer: THP-1 cells; Lower layer: OE-NC treated NCM460 cells). A minimum of three separate experiments were carried out and the data presented are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 4.
THY1 promotes apoptosis in DSS-induced colitis through the Bax/cleaved caspase3/Bcl-2 pathway.
(A) Representative TUNEL staining images of DSS-induced colitis mouse in the Sham (blank control) group, the IBD model group, the IBD + OE-THY1 group, the IBD + OE-NC group. the IBD + KD-THY1 group, and the IBD + sh-NC group. (B) The expression levels of apoptosis-related proteins (Bax, Bcl-2 and cleaved caspase3) in DSS-induced colitis mouse colon tissue samples of the Sham (blank control) group, the IBD model group, the IBD + OE-THY1 group, the IBD + OE-NC group. the IBD + KD-THY1 group, and the IBD + sh-NC group. A minimum of three separate experiments were carried out and the data presented are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 5.
THY1 promotes apoptosis in intestinal epithelial cells through the Bax/cleaved caspase3/Bcl-2 pathway.
(A) Apoptosis level and (B) apoptosis-related proteins (Bax, Bcl-2 and cleaved caspase3) expression levels of NCM460 cells in co-culture system of different treatment groups were determined by flow cytometry and western blot analysis, respectively. A minimum of three separate experiments were carried out and the data presented are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.
Fig 6.
THY1 promotes angiogenesis via upregulation of HIF-1α and VEGF expression in IBD.
(A) Representative H&E staining images of DSS-induced colitis mouse colon samples revealed the growth and thickness of the blood vessel wall in different treatment groups. The red square position in the image revealed the blood vessel. (B) The mRNA expression levels and (C) protein expression levels of HIF-1α and VEGF in DSS-induced colitis mouse colon tissue samples determined by quantitative real-time PCR and western blot analysis, respectively. (D) The in vitro angiogenic capacity of HUVECs stimulated with the cell culture supernatants collected from different treatment groups. (E) The protein expression levels of HIF-1α and VEGF in NCM460 cells in co-culture system of different treatment groups were determined by western blot analysis. A minimum of three separate experiments were carried out and the data presented are expressed as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001.