6-羟基多巴定向注射建立帕金森病大鼠模型的实验研究

Experimental study on establishment of PD's rats model by 6-OHDA directional injection

目的探讨立体定向间隔注射6-羟基多巴(6-OHDA)毁损黑质致密部(SNc)和中脑腹侧被盖(VTA)建立类似于人类帕金森病(PD)中晚期的PD大鼠模型方法。方法近交系Wistar大鼠50只,脑立体定向将6-OHDA注入大鼠右侧VTA及SNc,间隔两周,对阿朴吗啡(Apo)诱发旋转后旋转不明显或无稳定左侧旋转模型再次制模,并观察大鼠行为学改变,免疫组化检测黑质多巴胺(DA)能神经元的数量以及高效液相-荧光法检测黑质纹状体中DA含量的变化。结果(1)50只大鼠中有41只经APO诱导表现为恒定左侧旋转且结果稳定,旋转圈数>210r/30min,视为成功PD大鼠模型,部分大鼠伴有震颤、活动迟缓、嗅探、觅食、竖尾等异常行为改变,并且可持续存在16周;(2)免疫组化结果:PD大鼠模型毁损侧黑质区多巴胺能神经元较对侧及对照组减少大于90%;(3)PD大鼠右侧黑质纹状体中DA含量较左侧及对照组减少90%以上。结论6-OHDA毁损SNc及VTA间隔注射法可有效建立模拟人类PD中晚期的大鼠PD模型。

Objective To study the methods of establishing rat subjects with Parkinson＇s disease （PD） by injecting 6-hydrodopamine （6-OHDA） occasionally and steric directionally into SNc and VTA, which, is similar to human PD of middle-latter period. Methods Subject of ambiguousness of rotating after being induced to rotate or unstable left rotating subject was made again two weeks after cerebral steric directional injection of 6-OHDA into VTA in the right and SNC or 50 Wistar rats of inbred line. The changes of behavior, neurons＇ quantity if DA tested by immunohistochemistry and the level of dopamine in nigrostriatal system were observed. Results （1） 41 of rats showed steadily apormorphine-induced rotation（82%） and they could last for 16 weeks. The rotation rate was above 210 r/30 min, which were regard as success PD rats, parts of which displayed abnormal behaviors, such as moving stiffly, sniffing, seeking for food, railerection, et al. （2） The result of immunohistochemistry： neuron in damaged side of DA was reduced by more than 90% compared with that in the opposite side and control group. （3） The level of DA in PD rats＇ right nigrostriatal system was obviously reduced compared to that of the control and PD rats＇ left side. Conclusion Unilateral interval injection of 6-OHDA into the SNc and VTA can establish PD rat models steadily and effectively, which is similar to human PD of middle-latter period.