CXCL16 protects against oxygen and glucose deprivation-induced injury in human microvascular endothelial cells-1: Potential role in ischemic stroke.

AIM

To explore the protective effect of chemokine ligand 16 (CXCL16) against cell damage induced by oxygen-glucose deprivation (OGD) in human microvascular endothelial cells-1 (HMEC-1) and its possible mechanism.

METHODS

Cell Counting Kit-8 (CCK-8) assay and flow cytometry were performed to determine cell viability and apoptosis of HMEC-1, respectively. qRT-PCR analysis was applied to display the expression of CXCL16 and miR-424. Western blot analysis was used to detect the expression of apoptosis-related proteins, CXCL16, cAMP/PKA/CREB, and PI3K-AKT-GSK3β pathway-related proteins.

RESULTS

OGD significantly inhibited cell viability and promoted apoptosis. CXCL16 overexpression decreased the proliferation inhibition and apoptosis of HMEC-1 induced by OGD. Furthermore, we found that CXCL16 was a target of miR-424 and was downregulated by miR-424. The further study showed that overexpression of miR-424 significantly increased proliferation inhibition and apoptosis of HMEC-1 induced by OGD. In addition, we also found that miR-424 was downregulated by PMS2L2. In the subsequence experiment, overexpression of PMS2L2 significantly decreased the proliferation inhibition and apoptosis of HMEC-1 induced by OGD, which suggested that PMS2L2 decreased cell damage of HMEC-1 induced by OGD. Simultaneously, CXCL16 treatment markedly increased the phosphorylation of PKA/CREB and PI3K-AKT-GSK3β and these signal pathways were blocked by signal inhibitors.

CONCLUSION

Our study first demonstrates that oxygen-glucose deprivation (OGD)-induced human microvascular endothelial cells-1 (HMEC-1) cell injury was alleviated by CXCL16 targeted by miR-424 which further targeted by PMS2L2. This process might also be regulated by activating PKA/CREB and PI3K-AKT-GSK3β pathways.