Thrombin activity enhancement and its receptor protease‐activated receptor 1 (PAR‐1) activation play vital roles in neurologic deficits in the central nervous system. Our recent study showed that PAR‐1 upregulation stimulated by chronic high glucose (HG) caused central neuron injury through neuroinflammation; however, the molecular mechanisms are far from clear. In the present study, we found that HG resulted in neuronal injury of SH‐SY5Y cells as evidenced by decreased cell viability and increased lactate dehydrogenase release and elevated the mRNA level of PAR‐1. Moreover, we predicted and determined several potential microRNAs (miRs) combining with the 3′‐UTR of PAR‐1 mRNA, finding that miR‐20a‐5p, miR‐93‐5p, and miR‐190a‐5p were significantly decreased in HG‐cultured SH‐SY5Y cells compared with control. Further, SH‐SY5Y cells stably transfected with miR‐20a‐5p or miR‐190a‐5p mimic were established, and overexpression efficiency were confirmed. It was found that miR‐20a‐5p or miR‐190a‐5p overexpression markedly decreased PAR‐1 mRNA level and protein expression in SH‐SY5Y cells cultured with HG and normal glucose, indicating that miR‐20a or miR‐19a deficiency contributed to HG‐induced PAR‐1 upregulation. Together, our findings demonstrated that PAR‐1 upregulation mediated HG‐induced neuronal damage in central neurons, which was achieved through miR‐20a or miR‐190a deficiency.