Synphilin-1寡核苷酸对6-羟多巴胺诱导帕金森病大鼠黑质Caspase-3表达影响

THE INFLUENCE OF Synphilin-1 OLIGONUCLEOTIDES ON THE EXPRESSION OF Caspase-3 IN THE SUBSTANTIA NIGRA IN RAT MODEL WITH PARKINSON DISEASE INDUCED BY 6-HYDROXYDOPAMINE

目的观察Synphilin-1寡核苷酸对6-羟多巴注(6-OHDA)诱导的帕金森病(PD)大鼠黑质Caspase-3蛋白表达的影响,从而探讨Synphilin-1在帕金森病发病机制中的作用。方法Wistar大鼠60只随机分成5组,PD组、假手术组、反义组、正义组、错义组,每组12只大鼠。PD组立体定向将6-OHDA注入左侧黑质制作PD模型,反义组、正义组及错义组分别立体定向将反义、正义、错义寡核苷酸注入到各组鼠的左侧黑质,然后用与PD模型组相同的方法注入6-OHDA,假手术组注入生理盐水作为对照。术后4周取大鼠的术侧中脑黑质,应用免疫组化方法检测Synphilin-1及Caspase-3阳性细胞的表达。结果与假手术组比较,PD组Synphilin-1、Caspase-3阳性细胞表达显著增加(F=16.6、9.4,q=4.21、6.79,P<0.05、0.01);与PD组比较,反义组Synphilin-1、Caspase-3阳性细胞表达显著减少(q=5.43、4.92,P<0.01);正义和错义组Synphilin-1、Caspase-3阳性细胞的表达与PD组比较未见明显差异(q=3.31、3.52,P>0.05)。结论Synphilin-1反义寡核苷酸对6-OHDA诱导的PD大鼠旋转行为有明显的改善作用,提示Synphilin-1反义寡核苷酸对黑质神经元损伤具有保护作用,这种保护作用可能至少部分是通过调节Caspase-3的表达来实现的。

Objective To observe the effects of Synphilin-1 on the expression of Caspase 3 in substantia nigra in rat with 6 hydroxydopamine （60HDA）-indueed Parkinson disease （PD）, and explore the role of Synphilin-1 in its nosogenesis. Methods Sixty Wistar rats were evenly randomized to five groups： i.e. sham operation, PD, antisense, sense, and missense. For the PD group： 6-OHDA was stereotaxically injected into the left substantia nigra to construct a PD model. For antisense, sense, and mis sense groups; antisense , sense , and missense-oligonucleotides were stereotaxically injected into the left substantia nigra, respectively, and then 60HDA was stereotaxically injected into the left substantia nigra with the same method used in the PD group. Those in the sham-operation group were injected with same amount of normal saline as the control. Four weeks after the operation, the rat brains were removed and the expressions of Synphilin 1, Caspase-3 were detected by immunohistochemistry. Results Compared with sham operation group, the expressions of Synphilin-1 and Caspase-3 significantly increased in PD group （F= 16.6, 9.4;q=4. 21,6. 79;P〈0.05,0. 01）; compared with PD group, the expressions of Synphilin-1 and Caspase-3 significantly de creased in antisense group （q = 5.43,4.92 ; P〈0.01）. The expressions of Synphilin 1, Caspase-3 in the sense and missense groups had no significant difference compared with PD group （q=3.31,3.52;P〉0.05）. Conclusion Synphilin-1 antisense oligonucleotides obviously improve circling behavior caused by 60HDA, indicating Synphilin-1 antisense oligonucleotides have a neuroprotective effect against substantia nigra neurons damage. This protection is achieved, at least, by regulating the expression of Caspase-3.