不同部位注射6-羟基多巴胺建立帕金森病大鼠模型比较分析

Comparative Analysis on Rat Models of Parkinson's Disease Established by Injecting 6-hydroxy Dopamine with Different Sites

目的探索6-羟基多巴胺(6-OHDA)脑内不同部位注射建立稳定、有效的帕金森病(PD)大鼠模型的方法。方法 SD大鼠随机分为中脑单点组、中脑双点组、纹状体单点双注组及对照组,将6-OHDA按照分组方案注入大鼠脑内,分别在注射后1周、2周、4周、8周观察大鼠的行为学变化;8周取大鼠脑组织进行病理学检测并对络氨酸羟化酶(TH)免疫组织化学阳性细胞进行分析。结果 (1)中脑单点组造模成功率为72.7%,中脑双点组为60.0%,纹状体单点双注组为47.6%,对照组为0%(P<0.05)。(2)中脑单点组第1周时PD模型大鼠平均旋转圈数为(7.3±0.2)r/min,2周时较前增加(P<0.05),4周时较前增加(P<0.05),8周时有所增加但无差别(P>0.05);中脑双点组和纹状体单点双注组平均旋转圈数亦有相似变化趋势,但相同时间点各组间平均旋转圈数无差异(P>0.05)。(3)中脑单点组、中脑双点组及纹状体单点双注组PD大鼠损毁侧TH阳性细胞平均积分光密度(IOD)明显比对照组小(P<0.01);中脑单点组PD大鼠损毁侧TH阳性细胞IOD比中脑双点组及纹状体单点双注组小(P<0.05)。结论 6-OHDA中脑黑质致密区单点注射法建立的PD大鼠模型成功率高,模型稳定,死亡率低。

Objective To explore the way of establishment of stable and effective Parkinson＇s Disease（PD） rats model with 6-hydroxyl dopamine（6-OHDA）. Methods Firstly, SD rats were randomly divided into four groups, the one spot via midbrain group, the double spots via midbrain group, the one spot-twice injections via striatum group and control group. And 6-OHDA was injected into the brain of rats respectively according to different groups. Then observed the behavioral changes of rats respectively in the first week, second week, fourth week and eighth week, and tissue of brain was obtained after observation of behavioral changes in the eighth week, and then pathological examination was performed by HE staining and immunohistochemistry. Finally, tyrosine hydroxylase（TH） positive cells were analyzed with biosens digital imaging system. Results（1） The success rate of PD rats of the one spot via midbrain group was 72.7%, the double spots via midbrain group was 60%, the one spot-twice injections via striatum group was 47.6%, and control group was 0%. The success rates of different groups were statistically significant（P〈0.05）.（2）The average revolving number of successful PD model of the one spot via midbrain group in the first week was 7.3 ± 0.2 r/min, and increased than the first week in the second week（P〈0.05）, increased than the second week in the fourth week（P〈0.05）,and increased than the fourth week in the eighth week, but there was no statistically significant（P〈0.05）. There was the similar behavioral change trend in the double spots via midbrain group and the one spottwice injections via striatum group, but there was no statistical significance at the same time among different groups（P〉0.05）.（3） After analysis of TH positive cells of the damaged side of PD rats with biosens digital imaging system, the average integral optical density（IOD） could be obtained, and the IOD of the one spot via midbrain group, the double spots via midbrain group and the one spot-twice injections via striatum group was significantly smaller than that of the control group（P〈0.01）, and the IOD of the one spot via midbrain group was smaller than that of the double spots via midbrain group and the one spot-twice injections via striatum group（P〈0.05）. Conclusions The results suggest that after injection of 6-OHDA into the substantia nigra pars compact singly, the success rate of PD rats is higher, the PD rats is more stable and the mortality rate is lower.