The impact of oxygen-glucose deprivation/reperfusion on the Wnt5a/FZD2/Ca²⁺ pathway in neurons

Objective  To investigate the changes in the Wnt5a/FZD2/Ca²⁺pathway in neuronal cells after oxygen glucose deprivation reperfusion (OGD/Rep).

Methods  The OGD/Rep model was used to construct neuronal cell local ischemia/reperfusion injury, and cells were divided into normal group and reperfusion (0, 3, 6, 9, 12, 24 h) group.Cortical neuron cells was identified by immunofluorescence.The OGD time was determined by CCK-8 method.Western blotting was used to detect the changes of Wnt5a, FZD2, IP3-R, and p-CaMKⅡ/CaMKⅡ proteins in non-classical Wnt/Ca²⁺ signaling pathways.Immunofluorescence and enzyme-linked immunosorbent assay (ELISA) were used to detect the levels of intracellular Ca²⁺ and ROS.

Results  MAP-2 immunofluorescence identification showed that the purity of primary cortical neurons was relatively high.Compared with the normal group, the survival rate of neuronal cells after 3 hours of OGD was 55.46%.Compared with the normal group, the relative expression levels of non classical Wnt/Ca²⁺ signaling pathway related proteins Wnt5a (3, 6, 9, 12, 24 h), FZD2 (3, 6, 9 h), IP3-R (6, 9, 12, 24 h), and p-CaMKⅡ/CaMKⅡ (6 h) were upregulated after OGD/Rep (P < 0.05 to P < 0.01).Compared with the normal group, the expression levels of Ca²⁺ and ROS in nerve cells increased after OGD/Rep (P < 0.01).

Conclusions  OGD/Rep in neuronal cells can activate the Wnt5a/FZD2/Ca²⁺signaling pathway.