石杉碱甲对血管性痴呆小鼠海马神经细胞[Ca~(2+)]_i及钙调蛋白、蛋白激酶Ⅱ信使核糖核酸表达的影响

目的 :观察石杉碱甲对血管性痴呆 (VD)小鼠海马神经细胞 Ca2 + ] i 和钙调蛋白 (CaM) ,Ca2 +钙调蛋白依赖性蛋白激酶Ⅱ (CaMPKⅡ )mRNA表达的影响。方法 :采用双侧颈总动脉线结法 ,制作小鼠VD模型 ,并设假手术对照组 ,石杉碱甲为治疗组 ;术后d 2 9,30测试学习、记忆成绩。利用激光共焦显微镜检测各组海马神经细胞 Ca2 + ] i;用RT PCR技术检测CaM ,CaMPKⅡmRNA。结果 :模型组Ca2 + ] i 荧光强度 (44±s 3)显著高于假手术组(2 6± 4) (P <0 .0 1 )和石杉碱甲组 (2 8.5± 2 .5) (P<0 .0 1 ) ;模型组CaMmRNA含量 (0 .76± 0 .2 1 )显著低于假手术组 (1 .1 3± 0 .2 3) (P <0 .0 1 )和石杉碱甲组 (0 .97± 0 .1 9) (P <0 .0 5) ,CaMPKⅡmRNA含量(0 .43± 0 .0 7)显著低于假手术组 (0 .67± 0 .1 0 )(P <0 .0 1 )和石杉碱甲 (0 .61± 0 .0 8) (P <0 .0 1 )。结论 :石杉碱甲可降低VD小鼠海马神经细胞Ca2 + ] i,提高CaM ,CaMPKⅡmRNA表达水平 ,而改善VD临床症状

Effects of huperzine A on [Ca2+]i level and expression of CaM,CaMPKⅡ mRNA in hippocampal neurons of mice with vascular dementia

AIM: To investigate the effects of huperzine A on the Ca 2+ ] i level and exp ression of calmodulin ( CaM),calmodulin-dependent protein kinase Ⅱ(CaMPKⅡ)mRNA in hippocampal neuro n s of the mice with vascular dementia (VD). METHODS: The mice were subjected to repeated ischemia-reperfusion on bilateral common carotid arteries by knots to establish the VD models. Those animals with the shamed-operation were taken as control group. In treating group, huperzine A was administrated to the mice with VD. The changes of behavior were observed t hrough the step-down avoidance test and water maze test on the d 29, 30 after operations. The Ca 2+ ] i level of hippocampal neurons was evaluated by laser scan ning confocal microscopy. RT-PCR technique was used to measure the expression o f CaM,CaMPKⅡmRNA in hippocampal neurons. RESULTS: The Ca 2+ ] i level was apparently higher in model group(44±s 3) than that in the shamed-oper ation group (26±4) (P<0.01) and treating group(28.5±2.5)(P< 0.01) , and revealed no significant difference between shamed group and treating group . The expression of CaM mRNA (0.76±0.21) in VD group was statistically lower t han that in control group(1.13±0.23)(P<0.01) and treating group (0.97±0 .19)( P<0.05), CaMPKⅡmRNA in VD group(0.43±0.07) was statistically lower than that in control group(0.67±0.10)(P<0.01)and treating group (0.61±0.08) (P <0.01); there was no significant difference between control group and treating group. CONCLUSION: Huperzine A can redu ce Ca 2+ ] i level, increase the expression of CaM,CaMPKⅡmRNA , and furthermore, im prove the clinical symptom of VD.