VANGL2 inhibits NLRP3 inflammasome activation and DSS-induced colitis progression through the MARCH8 E3 enzyme activity-dependent pathway.

(A) Alcian blue staining was used to detect the intestinal characteristics of WT mice, <i>Vangl2</i>^ΔM/ΔM mice, and MCC950 treated <i>Vangl2</i>^ΔM/ΔM mice after DSS treatment. Scale bar = 100 μm. (B) Inflammasome-related plasmids (NLRP3, ASC, pro-Casp1, and IL-1β), HA-VANGL2, and GFP-MARCH8 (WT, W114A) plasmids were transfected into HEK293T cells (<i>MARCH8</i> KO) for 24 h, and the expression of IL-1β was detected by ELISA. (C–I) The construction of March8-WT, March8-W110A, or Vangl2 overexpression lentiviral plasmids and virus packaging were used to infect <i>March8</i> KO bone marrow (BM) cells according to the following 4 groups: March8-WT, March8-WT+Vangl2, March8-W110A, March8-W110A+Vangl2. Recipient mice aged 6–8 weeks were irradiated with 4.5 Gy, then irradiated again with 4 Gy 4 h later, and randomly divided into 4 groups to receive lentivirus-treated BM cells, respectively. Chimeric mice were kept in SPF condition and used for the construction of DSS-mediated colitis after 6–8 weeks. The survival rate (C), weight change (D), disease activity index (E), colon length (F and G), HE staining of colon (H), and colon IL-1β (I) were measured. Data are expressed as means ± SD. *<i>P</i> < 0.05, ***<i>P</i> < 0.01, ***<i>P</i> < 0.001, NS means not significant. (PDF)