Esketamine promotes proliferation and differentiation of mouse osteoblasts through NMDA receptor
Objective To explore the effect and its mechanism of esketamine on the proliferation and differentiation of osteoblasts.Methods Osteoblasts were extracted and transformed from C57B 1/6 male mice.The toxicity of different concentrations(0,6.25,12.5,25,50,100,200 µmol/L)of N-methyl-D-aspartic acid(NMDA)for 24,48,72,96 h to osteoblasts was detected by cell counting kit-8(CCK-8)method to screen the appropriate concentration.The induced osteoblasts were divided into three groups:control group,esketamine(Ket)group and Ket+NMDA group.After 24,48,72 h intervention,the mRNA expression levels of Runt associated tran-scription factor 2(Runx2)and the osteoblast specific transcription factor Osterix in each group were detected by real-time fluorescence quantitative PCR(qPCR),and the expression levels of B-lymphoblastoma-2 gene(Bcl-2),Bcl-2-associated X gene(Bax)and brain-derived neurotrophic factor(BDNF)proteins in osteoblasts in each group were determined by enzyme-linked immunosorbent assay(ELISA).Results CCK-8 results showed that 0-200 µmol/L NMDA had no obvious toxicity to osteoblast proliferation at the same time point(P＞0.05)and there was no significant difference in the activity of osteoblasts after NMDA intervention for different time(P＞0.05).The qPCR results showed that the expression levels of Runx2 and Osterix in Ket group and Ket+NMDA group were higher than those in control group(P＜0.01)and the expression levels of Runx2 and Osterix in Ket+NMDA group were lower than those in Ket group(P＜0.01).ELISA results showed that the expression levels of Bcl-2 protein and BDNF protein in Ket group were higher than those in control group,while the expression level of Bax protein was lower(all P＜0.05).Compared with Ket group,the expression levels of Bcl-2 protein and BDNF protein were decreased and Bax protein was increased in Ket+NMDA group(all P＜0.05).Conclusion Esketamine may promote the proliferation and differentiation of mouse osteoblasts by blocking N-methyl-D-aspartate receptor(NMDAR),which increases BDNF expression,thus increasing the expression of anti-apoptotic proteins while inhibiting the expression of apoptotic proteins.
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