论文部分内容阅读
目的探讨TRPC离子通道通过p53通路参与一氧化氮供体硝普钠(SNP)诱导的原代培养海马神经元凋亡。方法原代培养的海马神经元,经RT-PCR检测TRPC1-6的表达;给予SNP处理后,通过Real-time RT-PCR检测p53下游靶基因Bax、Apaf-1、PUMA和Survivin的表达;给予TRPC通道阻断剂和激活剂,检测其是否影响SNP诱导的p53下游靶基因表达。结果 TRPC在原代培养海马神经元上广泛表达。SNP处理神经元4、8、24h后,p53下游靶基因Bax、Apaf-1和Survivin表达无明显变化(P>0.05)。而PUMA则在3个时间点分别增加至1.94、1.86和1.90倍,与对照组比较差异有统计学意义(P<0.01)。TRPC通道阻断剂2-APB,可降低SNP诱导的PUMA上调(1.91vs1.12)(P<0.01)。结论 TRPC通道可以通过调节PUMA的表达参与SNP诱导的原代培养海马神经元凋亡。
Objective To investigate the role of TRPC channels in the apoptosis of primary cultured hippocampal neurons induced by nitric oxide donor sodium nitroprusside (SNP) via the p53 pathway. Methods The primary cultured hippocampal neurons were detected by RT-PCR. The expression of TRPC1-6 was detected by Real-time RT-PCR and the expression of Bax, Apaf-1, PUMA and Survivin were detected by Real-time RT- TRPC channel blockers and activators to determine whether it affects SNP-induced p53 downstream target gene expression. Results TRPC was widely expressed in primary cultured hippocampal neurons. After SNP treatment for 4, 8 and 24 hours, the expressions of Bax, Apaf-1 and Survivin, a target gene of p53, had no significant changes (P> 0.05). While PUMA increased to 1.94, 1.86 and 1.90 times respectively at 3 time points, which was significantly different from the control group (P <0.01). TRPC channel blocker 2-APB reduced SNP-induced PUMA up-regulation (1.91 vs 1.12) (P <0.01). Conclusion TRPC channels can participate in SNP-induced apoptosis of primary cultured hippocampal neurons by regulating PUMA expression.