Aim To explore the effect and molecular mechanism of procaine on hypoxia induced injury of N9 and PC12 cells. Methods N9 cells and PC12 cells were divided into control group, hypoxia group, low, medium and high dose (hypoxia＋2,6, 18 mg/L procaine) procaine group, anti-miR-con group, anti-miR-369-3p group, miR-con＋high dose procaine group (transfection miR-con＋hypoxia＋18 mg/L procaine), miR-369-3p＋high dose procaine group (transfection miR-369-3p＋hypoxia＋18 mg/L procaine). Flow cytometry was used to detect apoptosis, reagent kit was used to detect the content of reactive oxygen species (ROS) and malondialdehyde (MDA) and the activity of superoxide dismutase (SOD), ELISA was used to detect the levels of inflammatory factors such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin-6 (IL-6). RT-qPCR was used to detect the expression of miR-369-3p. Results Compared with the control group, the apoptosis rate of N9 cells and PC12 cells in the hypoxia group increased, the content of MDA and ROS increased, the activity of SOD decreased, the levels of TNF-α, IL-1β and IL-6 increased, the expression of miR-369-3p increased (P＜0.05). Compared with the hypoxia group, the apoptosis rate of N9 cells and PC12 cells in the low, medium and high dose procaine groups decreased, the content of MDA and ROS decreased, the activity of SOD increased, the levels of TNF-α, IL-1β and IL-6 decreased, the expression of miR-369-3p decreased in a concentration dependent manner (P＜0.05). Compared with the anti-miR-con group, the apoptosis rate of N9 cells and PC12 cells in the anti-miR-369-3p group decreased, the content of ROS and MDA decreased, the activity of SOD increased, the levels of TNF-α, IL-1β and IL-6 decreased (P＜0.05). Compared with miR-con＋high dose procaine group, the apoptosis rate of N9 cells and PC12 cells in miR-369-3p＋high dose procaine group increased, the expression of cleaved Caspase-3 protein increased, the content of MDA and ROS increased, the activity of SOD decreased, the levels of TNF-α, IL-1β and IL-6 increased (P＜0.05). Conclusion Procaine can reduce hypoxia induced injury to N9 and PC12 cells, and its mechanism may be related to inhibiting the expression of miR-369-3p.