Role of hsa_circ_0025853 in hypothermia-induced reduction of oxygen-glucose deprivation and restoration injury to neuroblastoma cells / 中华麻醉学杂志

ABSTRACT

Objective:To evaluate the role of hsa_circ_0025853 in hypothermia-induced reduction of oxygen-glucose deprivation and restoration (OGD/R) injury to neuroblastoma cells (SK-N-SH cells). Methods:SK-N-SH cells were cultured in vitro to the logarithmic phase and divided into 4 groups ( n=20 each) using a random number table method: control group (group C), group OGD/R, hypothermia group (group H) and hsa_circ_0025853 small interfering RNA (siRNA) plus hypothermia group (group S+ H). The cells were cultured in normal culture atmosphere in group C. The cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply for 24 h in group OGD/R.The cells were subjected to OGD for 3 h followed by restoration of oxygen-glucose supply for 24 h at 32 ℃ in group H. The cells were transfected with siRNA at 48 h before establishing OGD/R model, and the other treatments were similar to those previously described in group H. At 24 h of restoration of oxygen-glucose, cell viability was measured by Cell Counting Kit-8, apoptosis rate were measured by flow cytometry.the expression of dynamin-related protein 1 (Drp1) mRNA and hsa_circ_0025853 was detected by quantitative real-time-polymerase chain reaction, and the expression of Drp1 protein was detected by Western blot. Results:Compared with group C, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of hsa_circ_0025853 was down-regulated, and the expression of Drp1 protein and mRNA was up-regulated in the other 3 groups ( P<0.05). Compared with group OGD/R, the cell viability was significantly increased, the apoptosis rate of cells was decreased, the expression of hsa_circ_0025853 was up-regulated, and the expression of Drp1 protein and mRNA was down-regulated in H and S+ H groups ( P<0.05). Compared with group H, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of hsa_circ_0025853 was down-regulated, and the expression of Drp1 protein and mRNA was up-regulated in group S+ H ( P<0.05). Conclusion:The mechanism by which hypothermia alleviates the OGD/R injury to SK-N-SH cells is related to the up-regulation of hsa_circ_0025853 expression, thus reducing the expression level of Drp1.