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目的检测Sonic Hedgehog蛋白(Shh)在帕金森病(Parkinson’s disease,PD)大鼠中脑黑质的表达。方法将健康SD大鼠(雌雄不拘)随机分成3组:正常组大鼠不实施任何处理;应用脑立体定位技术,在大鼠右侧纹状体两针道内4点定位注射含0.2g/L抗坏血酸的生理盐水4μL(假手术组)或6-羟基多巴(6-OHDA)12μg/4μL(模型组)。术后1、2、4、6周以阿朴吗啡检测其异常旋转行为,左侧旋转的圈数大于7r/min者为合格的帕金森病模型大鼠。6周后在激光共聚焦显微镜下观察黑质Shh及酪氨酸羟化酶(TH)的免疫荧光双重染色结果,用Leica Confocal Software对Shh的荧光强度进行定量分析。结果Shh蛋白在正常对照组和假手术组大鼠中脑黑质中无明显表达,而在模型组表达显著。结论PD模型大鼠黑质中Shh蛋白的明显表达,提示Shh在PD模型大鼠中脑多巴胺能神经元(DA神经元)修复过程中可能发挥着一定的作用。
Objective To detect the expression of Sonic Hedgehog protein (Shh) in substantia nigra of Parkinson’s disease (PD) rats. Methods Healthy SD rats were randomly divided into three groups: normal rats were not treated with any treatment; brain stereotactic technique was used in the right striatum in the two-track intranasal injection containing 0.2g / L Ascorbate normal saline 4 μL (sham operation group) or 6-OHDA (12 μg / 4 μL model group). At 1, 2, 4, and 6 weeks after surgery, apomorphine was used to detect the abnormal rotation behavior, and the number of rotations on the left side was more than 7r / min. Six weeks later, immunofluorescence double staining of Shh and tyrosine hydroxylase (TH) of substantia nigra was observed under confocal laser scanning microscope, and the fluorescence intensity of Shh was quantitatively analyzed by Leica Confocal Software. Results Shh protein was not expressed in midbrain substantia nigra of rats in normal control group and sham operation group, but significant in model group. Conclusions The Shh protein expression in the substantia nigra of PD model rats suggests that Shh may play a role in the repair of dopaminergic neurons (DA neurons) in PD model rats.