摘要
目的探讨miR-133b在甲基苯丙胺(methamphetamine,MA)依赖大鼠脑内表达情况及其对神经元毒性损伤的调控作用。方法通过腹腔内连续注射MA(10mg/kg),建立条件性位置偏爱(conditionedplacepreference,CPP)大鼠模型,应用实时荧光定量PCR(Real—timePCR,RT—PCR)技术检测模型鼠脑皮层miR-133b的表达变化。体外培养PCI2细胞并给予800μmol/LMA处理,RT—PCR检测miR-133b在培养神经细胞中的表达,并分别转染miR。133b模拟物和抑制物,观察miR-133b干扰后MA对培养神经细胞线粒体膜电位(mitochondfialmembranepotential,MMP)的影响。结果连续14d腹腔内注射MA后,大鼠在伴药箱停留时间[(620.20±44.80)s]明显长于对照组[(341.80±25.12)s,P〈0.01],表明成功建立了MA依赖鼠模型。RT—PCR检测发现MA模型鼠脑皮层miR-133b表达(0.36±0.05)较对照组(0.99±0.08)明显降低(P〈0.01)。在体外神经元模型中,PCI2细胞经MA处理后,可见大部分神经细胞胞体变圆,神经突起退缩,RT—PCR结果显示miR.133b表达(0.74±0.05)较对照组(1.00±0.02)降低(P〈0.05)。JC-1检测显示MA组[(109.85±7.03)个]MMP较对照组[(36.49±3.89)个]明显下降(P〈0.01),miR-133b模拟物组[(58.97±6.56)个]MMP较模拟物对照组[(135.46±15.04)个]明显增加(P〈0.01),miR-133b抑制物组[(162.84±14.15)个]MMP较抑制物对照组[(139.81±12.26)个]下降(P〈0.05)。结论miR-133b在MA依赖大鼠脑皮层组织和MA损伤体外神经元模型中表达均明显下调,通过调控miR-133b的表达,能够改变MA处理培养神经细胞的MMP损伤,表明miR-133b不仅参与了MA导致的神经损伤,还有望成为-个干预的分子靶点。
Objective To investigate the expression of miR-133h in the brain of methamphetamine (MA) dependent rats and its regulatory effects on neuronal toxic injury. Methods Through continuous in- traperitoneal injection to rats with MA( 10 mg/kg), the conditioned place preference (CPP) rats model was established,and the expression of miR-133b in the cerebral cortex of model rats was detected by real-time fluorescence quantitative PCR (RT-PCR). PC12 cells were cultured in vitro and treated with MA (800 μmol/L) ,and then miR-133b expression in cultured neurons was detected.miR-133b mimics and in- hibitor were transfected to PC12 cells respectively to observe the effect of miR-133b on the mitochondrial membrane potential(MMP) of cultured neurons. Results After continuous intraperitoneal injection with MAfor 14 days,the residence time of rats in the box with medicine((620.20±44.80) s) was significantly longer compared with the control group((341.80±25.12) s,P〈0.01 ) ,which showed that MA dependent rats model was successfully established.The RT-PCR detection results showed that the expression of miR-133b in the cerebral cortex of model rats( 0.36±0.05 ) significantly decreased compared with the control group (0.99± 0.08 ,P〈0.01 ).In the in vitro model, most of the neuronal cell bodies became round and the neuorites were withdrawn after MA treatment.Compared with the control group ( 1.00± 0.02 ) , the RT-PCR detection results showed that the expression of miR-133b in MA group(0.74±0.05) deereased(P〈0.05).The JC-1 detection results showed that the MMP of the MA group(109.85±7.03) decreased significantly contrast to the control group(36.49±3.89,P〈0.01) ,the MMP of the miPt-133b mimics group(58.97±6.56) increased significantly contrast to the mimics control group( 135.46±15.04,P〈0.01 ) and the MMP of the miR-133b inhibitor group (162.84 ± 14.15 ) decreased contrast to the inhibitor control group (139. 81 ±12. 26, P〈 0.05 ). Conclusions The expression of miR-133b in the cerebral cortex of MA dependent rats and in vitro neuron model treated with MA are significantly downregulated. By regulating the expression of miR-133b,the MMP damage of cultured neurons treated with MA is changed,indicating that miR-133b is not only involved in the nerve injury induced by MA, but also possiblely as a molecular target for intervention.
出处
《中华行为医学与脑科学杂志》
CAS
CSCD
北大核心
2017年第8期689-694,共6页
Chinese Journal of Behavioral Medicine and Brain Science
基金
基金项目:山东省医药卫生科技发展计划(2013WS0008)
山东省重点研发计划(2016GSF201054)
关键词
甲基苯丙胺
NMP
miR-133b
神经毒性
脑皮层
大鼠
Methamphetamine
Mitoehondrial membrane potential(MMP)
miR-133b
Neu- rotoxieity
Cerebral cortex
Rat