酸性鞘磷脂酶活化在微波辐照致神经细胞凋亡中的作用研究

Roles of acid sphingomyelinase activation in neuronal cells apoptosis induced by microwave irradiation

为阐明微波辐照致神经细胞酸性鞘磷脂酶(Acid Sphingomyelinase,ASM)活化的规律,揭示ASM活化在微波辐照致神经细胞凋亡中的作用,以原代培养神经元细胞,采用30W/cm2微波辐照10min,采用Hoechst 33342荧光染色观察细胞凋亡,以ASM活性测定试剂盒测定ASM活性,RT-PCR、western blot分别检测ASM基因和蛋白表达的变化。用ASM活性抑制剂米帕明预处理,观察微波辐照后神经细胞凋亡情况。研究发现,微波辐照引起海马神经元细胞发生凋亡,神经元ASM mRNA和蛋白表达上调,ASM酶活性增高,给予ASM活性抑制剂米帕明预处理后,可部分拮抗微波辐照引起的神经细胞凋亡率增加。结果提示,微波辐照引起神经细胞ASM活化及表达上升,ASM活化在微波辐照所致的神经细胞凋亡中起重要作用。

The present study is to examine the effect of microwave on acid sphingomyelinase （ASM） activity and expression, and to explore the role of ASM activation in neuronal ceils apoptosis induced by microwave irradiation. Primary cultured hippocampal neurons were irradiated by 30 W/cm2 microwave for 10 min, and ASM activity assay was used to investigate ASM activity alteration. RT-PCR and western blot were used to detect ASM mRNA and protein expression respectively. Apoptosis was observed by Hoechst 33342 fluorescence staining. ASM specific in- hibitor imipramine was applied to inhibit ASM activation. It has been found that apoptosis rate of primary cultured hippocampal neurons increased significantly after microwave irradiation. ASM was activated while ASM mRNA and protein expression were up-regulated in neurons after microwave irradiation. Pretreatment with imipramine could reverse neuronal apoptosis induced by microwave irradiation. Results show that microwave irradiation causes increment of ASM activation and expression and ASM activation is involved in microwave induced neuronal apoptosis.