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目的研究caspase-3反义寡核苷酸对6-OHDA诱导大鼠中脑多巴胺神经元凋亡的保护作用。方法向四组SD大鼠的中脑黑质部位注入反义、错义、正义寡核苷酸及NS,然后再注入6-OHDA,取中脑做连续切片,原位杂交及免疫组化检测黑质caspase-3的表达及TH的表达,原位末端标记法(Tunel)检测黑质细胞的凋亡。结果反义组Tunel阳性细胞数为82±8.6,方差分析显示反义组阳性细胞数显著性低于其他各组(p<0.05);反义组手术侧TH阳性细胞数为168.6±11.4,与对侧阳性细胞比值为63%±11.3%,显著性高于其余各组(P<0.05)。结论有效阻断caspase-3的表达可减轻6-OHDA诱导的多巴胺神经元凋亡,caspase-3可作为保护性治疗帕金森病的靶点。
Objective To study the protective effects of caspase-3 antisense oligonucleotide on apoptosis of dopaminergic neurons in rat midbrain induced by 6-OHDA. Methods The antisense, missense, sense oligonucleotide and NS were injected into the substantia nigra of middle cerebral artery of SD rats, and then injected into 6-OHDA. The midbrain was used for serial sections, in situ hybridization and immunohistochemistry The expression of substantia nigra caspase-3 and the expression of TH, the apoptosis of nigral cells were detected by Tunel. Results The number of Tunel positive cells in antisense group was 82 ± 8.6. ANOVA showed that the number of positive cells in antisense group was significantly lower than that in other groups (p <0.05). The number of TH positive cells in antisense group was 168.6 ± 11.4 The contralateral positive cells ratio was 63% ± 11.3%, significantly higher than the other groups (P <0.05). Conclusion The effective blocking of caspase-3 expression can reduce 6-OHDA-induced apoptosis of dopaminergic neurons and caspase-3 can be used as a protective target of Parkinson’s disease.