ZMS regulation of M_2 muscarinic receptor stability mediated by de novo synthesis of protein / 上海交通大学学报（医学版）

ABSTRACT

Objective To explore the mechanism of ZMS regulation of M_2 muscarinic receptor mRNA expression. Methods In vitro cultured CHOm2 cells were divided into ZMS 1 group (treatment with 1 × 10~(-5) mol/L ZMS for 24 h), ZMS 2 group (treatment with 1 × 10~(-5) mol/L ZMS for 24 h and 1 μg/mL cycloheximide for 12 h) and ZMS 3 group (treatment with 1 μg/mL cycloheximide for 4 h and 1 × 10 ~(-5) mol/L ZMS for 24 h), and their corresponding control groups were also established (substitution of ZMS by DMSO). Actinomycin D was added to cultured CHOm2 cells of each group to inhibit the synthesis of mRNA. CHOm2 cell samples were taken at different time points, the relative expression of M_2 receptor mRNA was detected by Real-time PCR, and half life of M_2 receptor mRNA was calculated. Results Compared with corresponding control groups, the half life of M_2 receptor mRNA of CHOm2 cells in ZMS 1 group and ZMS 2 group was significantly prolonged [(4.75h± 0.54) h vs (2.13 ±0.23) h, P<0.05; (5.43 ±1.13) h vs (2.46 ±0.09) h, P<0.05]. There was no significant difference in half life of M_2 receptor mRNA of CHOm2 cells between ZMS 3 group and its corresponding control [ ( 3.06 ±0.23) h vs (3.00 ± 0.20) h, P > 0.05]. Conclusion De novo protein synthesis is required for the enhancement of M_2 receptor mRNA stability regulated by ZMS.