Expression of p75 neurotrophin receptor in human neuroblastoma cells under condition of oxygen-glucose deprivation and its significance / 第二军医大学学报

ABSTRACT

Objective: To explore the expression and effect of p75 neurotrophin receptor (p75NTR) in human neuroblastoma cell line (SH-SY5Y cells) under the condition of oxygen-glucose deprivation (OGD). Methods: The OGD model of SH-SY5Y cells was established by glucose-free and serum-free culturing using tri-gas incubator, and then was assigned to 3 groups, including serum-free regular culturing group (control group), OGD group and OGD + LM11A-31 (a competitive blocker of p75NTR) group. After 12 h of culturing, the cell viability was measured by methyl thiazolyl tetrazolium (MTT) assay, lactate dehydrogenase (LDH) release activity was determined by LDH activity assay kit, cell apoptosis proportion was detected by flow cytometry, and p75NTR protein expression was detected by Western blotting. Results: The OGD model of SH-SY5Y cells was successfully established. Twelve hours after cell culture, the cell viability of the control, OGD and OGD + LM11A-31 groups was (94.80 ± 4.06)%, (50.34 ± 5.55)% and (64.68 ± 4.59)%, the LDH release activities were (46.93 ± 5.49) U/L, (353.09 ± 30.67) U/L and (282.20 ± 25.60) U/L, the proportions of apoptosis cells were (1.82 ± 0.45)%, (14.98 ± 2.59)% and (7.36 ± 1.98)%, and the relative expression levels of p75NTR were 0.06 ± 0.01, 0.41 ± 0.02 and 0.19 ± 0.03, respectively, and the differences were all significant (F=67.94, 142.10, 36.28 and 221.20, all P<0.05). Post hoc analysis showed that the cell viability of the OGD group was significantly lower than that of the control group, and the LDH release activity, the proportion of apoptosis cells and the relative protein expression level of p75NTR of the OGD group were significantly higher (Bonferroni test, all P' < 0.05). After treatment with LM11A-31, the cell viability of the OGD + LM11A-31 group was significantly higher than that of the OGD group, and the LDH release activity, the proportion of apoptosis cells and the relative protein expression level of p75NTR of the OGD LM11A-31 group were significantly lower (Bonferroni test, all P' < 0.05). Conclusion: The expression of p75NTR is increased in human neuroblastoma cell line SH-SY5Y cells under OGD condition, which may promotes neuronal injury and apoptosis.