摘要
目的 初步探讨苯丙胺（amphetamine,AMPH）模型大鼠海马的微小核糖核酸（microRNA）的表达情况.方法 应用microRNA微阵列芯片技术筛选AMPH模型大鼠（AMPH模型组,10只）和对照大鼠（对照组,10只）海马差异表达microRNA；选取有明显表达变化的microRNA,采用实时定量聚合酶链反应、免疫印迹法验证相关靶基因及蛋白在AMPH模型大鼠海马的表达变化.结果 （1）旷场实验：AMPH模型组水平运动格子数（28.21 ±2.22）个,垂直运动次数（82.33 ±4.61）次,均高于对照组[（17.10±1.94）个、（52.32±3.76）次],差异有统计学意义（t=11.92,P＜0.01;t=15.95,P＜0.01）.（2） AMPH模型组共有136个microRNA表达,其中,上调＞2倍的microRNA有14个,下调＞2倍的microRNA有6个；上调＞5倍的microRNA有3个,分别为microRNA-134、microRNA-143、microRNA-96,下调＞5倍的microRNA有2个,分别为microRNA-132、microRNA-210；实时定量聚合酶链反应验证了以上结果；通过microRNA特异性的靶标检测系统（miRanda）推测microRNA-134、microRNA-143、microRNA-96、microRNA-132、microRNA-210靶基因分别为谷氨酸受体1（GRM-1）、脑源性神经营养因子（BDNF）以及生长抑素（SSTR-1）.（3）免疫印迹法检测预测靶蛋白表达结果：AMPH模型组GRM-1为0.18±0.02、BDNF为0.21 ±0.02、SSTR-1为0.42 ±0.02,较对照组（分别为0.28±0.03、0.31±0.03、0.59±0.03）含量均下调,差异有统计学意义（t=8.77,P＜0.05；t=8.77,P＜0.05；t=14.91,P＜0.05）.结论 AMPH模型中海马microRNA呈差异表达.
Objective This study compared the difference of micro-ribonucleic acid （microRNA） expression in hippocampus of rat brain between an animal model of amphetamine（AMPH） and the normal by using the microRNA microaaray chip technology.Methods Twenty male SD rats were divided into control group（10） and AMPH group（10）.The differential expression of microRNA was screened by using the microRNA microaaray chip technology.The microRNAs and target proteins expression in the hippocampus in AMPH model were verified by real time PCR and western blot.Results （1） Numbers of crossings （28.21 ±2.22）and rearings（82.33 ±4.61）were significantly increased in AMPH groups compared with the control groups （crossings 17.10 ± 1.94 and rearings 52.32 ± 3.76） （t =11.92,P 〈 0.01 ; t =15.95,P 〈0.01）.（2） 136 microRNA were found in the hippocampus in AMPH group.Among those 14 raised up two times higher microRNAs and 6 lower two times microRNAs were found in the hippocampus in AMPH group.Three five times higher microRNAs were microRNA-134,microRNA-143 and microRNA-96,2 five times lower microRNAs were microRNA-132 and icroRNA-210.It was verified by real time PCR,the predicted target genes by miRanda algorithm of five times higher microRNAs were GRM-1,BDNF and SSTR-1.（3） The target genes were detected using Western blot,the relative contains of GRM-1 （0.18 ± 0.02）,BDNF （0.21 ± 0.02） and SSTR-1 （0.42 ± 0.02） in AMPH group were significantly lower than control group （0.28 ±-0.03,0.31 ±0.03,0.59 ±0.03）（t =8.77,P〈0.05; t =8.77,P〈0.05; t =14.91,P〈0.05）.Conclusion This findings indicate that there be microRNAs expression difference in AMPH model hippocampus.
出处
《中华精神科杂志》
CAS
CSCD
北大核心
2013年第5期301-305,共5页
Chinese Journal of Psychiatry
基金
广东省自然科学基金（S2011010001320）
广东省医学科研基金（B2010286）
深圳市科技计划项目（201102111）
深圳市心理健康重点实验室项目（201201）
