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目的探讨西洛他唑(CLZ)对乳鼠心室肌细胞(NRVMs)瞬时外向钾电流(Ito1)通道Kv4.3基因mRNA水平表达的影响及其治疗Brugada综合征(BrS)的分子机制。方法将培养的SD乳鼠心室肌细胞随机分为对照组及10、20、40、60、80、100μmol/LCLZ干预组,分别培养至24、48、72h,应用半定量逆转录-聚合酶链反应(RT-PCR)技术检测Ito1通道Kv4.3基因mRNA表达的变化。结果RT-PCR检测Kv4.3基因mRNA表达,不同浓度CLZ干预至24h,除10μmol/L组外,各组均较对照组显著降低(P<0.05);至48h,各干预组均较对照组呈浓度依赖性显著降低(P<0.05);而至72h,各干预组均较对照组显著增加(P<0.05)。结论CLZ与心室肌细胞共孵育,Kv4.3基因mRNA表达呈先下调后上调改变,提示CLZ可能适用于BrS的短期药物治疗,而非长期维持治疗。
Objective To investigate the effect of cilostazol (CLZ) on the expression of Kv4.3 mRNA in the transient outward potassium current (Ito1) channel of neonatal rat ventricular myocytes (NRVMs) and the molecular mechanism of treatment of Brugada syndrome (BrS). Methods SD rat neonatal rat ventricular myocytes were randomly divided into control group and 10, 20, 40, 60, 80, 100μmol / L LCZZ intervention groups and cultured for 24,48,72h respectively. Semi-quantitative RT- (RT-PCR) technique was used to detect the mRNA expression of Kv4.3 gene in Ito1 channel. Results The expression of Kv4.3 mRNA was detected by RT-PCR. The CLZ concentrations were significantly lower than those of the control group (P <0.05) except for 10μmol / L group (P <0.05). However, at 72h, the intervention groups were significantly increased compared with the control group (P <0.05). Conclusions CLZ co-incubated with ventricular myocytes, Kv4.3 mRNA expression was down-regulated after the first increase, suggesting that CLZ may be suitable for short-term drug treatment of BrS, rather than long-term maintenance treatment.