【摘要】 目的观察甲基苯丙胺(methamphetamine,METH)对大鼠海马神经元的损伤作用及炎性反应。方法原代培养大鼠胎鼠海马神经元,利用MTT和TUNEL实验技术观察METH对神经元的损伤;采用Affymetrix芯片结合实时荧光定量聚合酶链反应法(realtime-PCR)法,观察METH作用后海马神经元细胞炎性相关因子的表达变化。结果METH对海马神经元的损伤具有浓度依赖性。30μmol/L的METH即可引起神经元的活力降低。基因组芯片结果显示,与对照组相比,METH处理组与炎症信号相关,增加的有IL-1R、IL-1rap、IL-4、IL-10、IL-15、IL-24、Peli2、Peli3、NOS1、NOS2、Sig1-R、TNFα和TLR4;减少的有IL-1β、Peli1、Traf3、Traf6和NOS3。实时PCR结果显示,与对照组相比,METH使炎性因子IL-1R、IL-1rap、IL-4、IL-24、Peli1、Peli2、Peli3、Traf3、Traf6、NOS1、NOS2、NOS3、Sig1-R、TNFα和TLR4的mRNA表达增加,其中IL-1R、IL-1rap、IL-4、IL-24、Traf6和NOS3的表达增加有统计学意义(P<0.05);IL-1β、IL-6、IL-10、IL-15和Traf3的mRNA表达降低,其中IL-1β和Traf3的表达减少有统计学意义(P<0.05)。结论 METH可导致海马神经元损伤,影响炎性及相关因子的表达,可能会对学习记忆能力造成潜在影响。更多还原

【Abstract】 Objective To explore the adverse effects of methamphetamine( METH) on hippocampal neurons in rats and inflammatory reaction. Methods The neuronal damage induced by METH was detected by TUNEL and MTT assays after primary culture of rat fetal hippocampal neurons. Effects of METH on the expression of inflammatory cytokines in hippocampal neurons was detected by affymetrix chip combined with real time fluorescence quantitative polymerase chain reaction( realtime-PCR). Result METH obviously induced hippocampal neural damage in a dose-dependent manner. It decreased cell viability at a concentration of 30 μmol/L. Genome microarray result showed that compared with the control group,the METH treatment group displayed obvious changes in inflammatory signaling. The factors or corresponding proteins including IL-1 R,IL-1 rap,IL-4,IL-10,IL-15,IL-24,Peli2,Peli3,NOS1,NOS2,Sig1-R,TNF and TLR4 were significantly increased,while other inflammatory proteins such as IL-1,Peli1,Traf3,Traf6 and NOS3 were substantially decreased. Compared with the control group,realtime-PCR result showed that METH increased the mRNA expression of inflammatory factors such as IL-1 R,IL-1 rap,IL-4,IL-24,Peli1,Peli2,Peli3,Traf3,Traf6,NOS1,NOS2,NOS3,Sig1-R,TNF-α,Among them,the increase of IL-1 R,IL-1 rap,IL-4,IL-24,Traf6,NOS3 expression was statistically significant( P < 0. 05). Meanwhile,METH decreased the mRNA expression of inflammatory factors including IL-1β,IL-6,IL-10,IL-15 and Traf3 while the decrease of IL-1βand Traf3 expression was statistically significant( P < 0. 05). Conclusion METH can cause damage to the hippocampal neurons and affect the expression of inflammatory factors,which may affect the learning and memory ability.更多还原