apoptosis
To explore whether C1QTNF6 in leukocytes affects hypoxia-induced BBB
disruption through an inflammatory response, we established an in vitro
model in which OGD-exposed hCMEC/D3 cells were co-cultured with HL-60
cells. HL-60 cells in the different groups were treated as
aforementioned. Flow cytometry analysis revealed a decreased apoptosis
ratio in hCMEC/D3 cells in the OGD+H19 siRNA-HL-60 group than in the
OGD+control-HL-60 group (Figure 6A, B, P < 0.05). However,
there was an increased apoptosis ratio in hCMEC/D3 cells in the
OGD+miR-29b antagomir-HL-60 group than in the OGD+control-HL-60 group
(Figure 6A, B, P < 0.05). However, the increased hCMEC/D3 cell
apoptosis induced by OGD+miR-29b antagomir-HL-60 was reversed by H19
siRNA treatment (Figure 6A, B, P < 0.05).
Additionally, we measured C1QTNF6, IL-1β, and TNF-α expression in HL-60
cells and ZO-1 and occludin expression in hCMEC/D3 cells. Treatment with
miR-29b antagomir significantly increased C1QTNF6, IL-1β, and TNF-α
expression in HL-60 cells (Figure 6C, P < 0.05). There
was a significant decrease in C1QTNF6, IL-1β, and TNF-α expression in
HL-60 cells treated with H19 siRNA than in OGD HL-60 cells (Figure 6C,P < 0.05). Furthermore, we observed changes in ZO-1 and
occludin levels in hCMEC/D3 cells, as well changes in C1QTNF6, IL-1β,
and TNF-α expression in HL-60 cells. (Figure 6C). These findings suggest
that H19 in leukocytes aggravates hypoxia-induced hCMEC/D3 apoptosis by
targeting miR-29b/C1QTNF6.