DNMT3a-Mediated Enterocyte Barrier Dysfunction Contributes to Ulcerative Colitis via Facilitating the Interaction of Enterocytes and B Cells

Figure 5

DNMT3a regulates IEC-induced B cell differentiation through TNFSF13-mediated DNA methylation. (a) The si-DNMT3a or si-NC was transfected into IECs and then treated with LPS (1 μg/mL) for 48 h. Cells were grouped into 4 groups: IECs, IECs+LPS, IECs-si-NC+LPS, and IEC-si-DNMT3a+LPS groups. The methylation level and mRNA level in IECs and protein level of TNFSF13 in the supernatants were measured. (b) The si-DNMT3a and si-TNFSF13 were cotransfected into IECs and then treated with LPS (1 μg/mL) for 48 h. IECs were grouped as follows: IECs+LPS, IEC-si-DNMT3a+LPS, and IEC-si-DNMT3a-si-TNFSF13+LPS groups. The methylation level and mRNA level in IECs and protein level of TNFSF13 in the supernatants were measured. (c) IECs were cotransfected with si-DNMT3a and si-TNFSF13 and then cocultured with B cells. IECs were treated with LPS (1 μg/mL) for 48 h. The cocultural cells were assigned into 3 groups: IECs+B+LPS, IEC-si-DNMT3a+B+LPS, and IEC-si-DNMT3a-si-TNFSF13+B+LPS. Flow cytometry assay was conducted to detect the numbers of CD1d+ B cells and CD138+ B cells. (d) The inflammatory cytokines in cell supernatants were measured by ELISA. , vs. Blank or LPS group. Blank: IECs without any treatment; ^##, vs. LPS+si-NC or LPS+si-DNMT3a group.

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