Inhibition of microRNA-375 ameliorated ketamine-induced neurotoxicity in human embryonic stem cell derived neurons

Excessive exposure to commonly used anesthetic agents, such as ketamine, may induce permanent damage to immature human brains. In this work, we used a human embryonic stem cell (hESC)-derived neuron model to assess the expression and function of human microRNA 735 (hsa-miR-375) in regulating ketamine-induced neural cell death and neural toxicity in vitro. In the in vitro culture, hESC-derived neurons were incubated with ketamine for 72 h. After that, cell viability, reactive oxygen species activity, neural apoptosis, neurite degeneration, and hsa-miR-375 gene expression were assessed, respectively. We found ketamine induced neural death, reactive oxygen species augmentation, neural apoptosis, neurite degeneration and hsa-miR-375 upregulation, in hESC-derived neurons. In addition, we discovered that, lentivirus-mediated mR-375 downregulation protected ketamine-induced neural cell death and neural toxicity. Also, human brain derived neurotrophic factor (BDNF) was found to be directly and reversely regulated by hsa-miR-375. Moreover, BDNF downregulation was shown to functionally reverse the protective effect of miR-375 downregulation on ketamine-induced neural cell death and neural toxicity. Overall, this work provided strong evidence showing hsa-miR-375 is an active regulator in anesthesia-induced neural cell death and neural toxicity, possibly via inverse regulation on BDNF gene.