Effect of Ro20-1724 on ketamine-induced apoptosis in hippocampal neurons of neonatal rats / 中华麻醉学杂志

ABSTRACT

Objective To valuate the effect of Ro20-1724 on ketamine-induced apoptosis in hippocampal neurons of neonatal rats.Methods Hippocampal neurons from newborn Sprague-Dawley rats were obtained and cultured in vitro.The primary hippocampal neurons were randomly divided into 4 groups (n =24 each) using a random number tablecontrol group (group C),ketamine group (group K),solvent control group (group E),and Ro20-1724 group (R group).The neurons were incubated for 72 h in the normal culture medium in group C.The neurons were incubated for 72 h in the culture medium containing ketamine 150 μmol/L in group K.In E and R groups,after the neurons were incubated for 30 min in the culture medium containing ketamine 150 μmol/L,the culture medium was then replaced,0.01％ ethanol (final concentration) and 1 × 10-3 μmol/L Ro20-1724 (final concentration) were added to the culture medium,respectively,and the neurons were then incubated for 72 h.After 72 h incubation,the cell viability was detected by MTT assay,the cell apoptosis was detected by flow cytometry,the expression of Bcl-2 mRNA and Bax mRNA was determined by RT-PCR,and synaptophysin Ⅰ expression was detected by Western blot.The apoptosis rate was calculated.Results Compared with group C,the survival rate was significantly decreased,the apoptosis rate was increased,the expression of Bcl-2 mRNA and synaptophysin Ⅰ was down-regnlated,and Bax mRNA expression was up-regulated in K and E groups (P ＜ 0.05 or 0.01).Compared with group K,the survival rate was significantly increased,the apoptosis rate was decreased,the expression of Bcl-2 mRNA and synaptophysin Ⅰ was up-regulated,and Bax mRNA expression was downregulated in R group (P ＜ 0.05 or 0.01).Conclusion Ro20-1724 can inhibit ketamine-induced apoptosis in hippocampal neurons of neonatal rats and correction of Bcl-2/Bax imbalance is involved in the mechanism.