Silenced Notch1 attenuates autophagy in PC12 cells following oxygen-glucose deprivation-induced injury

Ischemic cerebrovascular disease (ICD) or ischemic stroke is one of the most severe and common neurological diseases in the elderly population. Autophagy is a highly conserved process in eukaryotes and a key decisionmaker contributing to neuronal fate. Until now, reports on the role of the Notch pathway in autophagy of ICD are limited. In this study, PC12 cells were screened for neuron-like cells that were positive for MAP2. The silenced Notch1 transfected neuron-like cells were used for the establishment of an oxygen glucose deprivation (OGD) model which was verified with cell viability by MTT assay and apoptosis examination by flow cytometry. After establishing the OGD model, the mRNA and protein expression of Notch1, Beclin1, and LC3-I/LC3-II were analyzed by RT-qPCR and Western blot. In the established OGD model, the cell viability was decreased significantly with 72.98% of apoptosis. Following the introduction of silenced Notch1, the mRNA and protein expression of Notch1, Beclin1 and LC3 were upregulated remarkably in neuron-like PC12 cells exposed to OGD compared with the control, increased markedly in Notch1 siRNA transfected neuron-like cells post-exposure to OGD compared with Notch1 siRNA transfected neuronlike cells without the treatment of OGD, and down-regulated distinctly in Notch1 siRNA transfected neuron-like cells post-exposure of OGD compared with those without transfection. We also observed that the levels of LC3-II rose greatly in OGD cells without Notch1 siRNA transfection compared with the control, but declined in OGD cells with Notch1 siRNA transfection compared with those without transfection. The Notch pathway might be partly associated with autophagy in neurons post-exposure to OGD highlighting it as a potential target for ICD treatment.