摘要
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 determine whether canonical transient receptor potential channel (TRPC) is involved in SNP- induced apoptosis of the primary cultured rat hippocampal neurons via PUMA. Methods The cultured rat hippocampal neurons were treated with SNP. The expression of TRPCI-6 in hippocampal neurons were detected using RT-PCR. The expression level of Bax, Apaf-1, survivin and PUMA mRAN in hippocampal neurons were detected using Real-time RT-PCR. Results TRPCs were widely expressed in hippocampal neurons. The hippocampal neurons were treated with SNP for 4 h, 8 h and 24 h, and SNP had no effect on the expression of Bax, Apaf-1 and survivin mRNA (P〉0.05) ; However, the expression level of PUMA was significantly increased to 1.94 (4 h), 1.86 (8 h) and 1.9-fold (24 h) (P〈0.01). TRPC blocker (2-APB) was found to lower the elevated expression of PUMA (P〈0.01). Conclusion TRPC channels are involved in SNP-induced apoptosis of cultured rat hippocampal neurons via PUMA.
出处
《热带医学杂志》
CAS
2012年第7期833-836,共4页
Journal of Tropical Medicine