“抗帕颗粒”对帕金森病模型鼠α-syn异常聚集的影响

摘要

目的探讨“抗帕颗粒”对1-甲基-4-苯基-1，2，3，6-四氢吡啶（MPTP）帕金森病（PD）模型小鼠黑质纹状体区α-突触共核蛋白（α-syn）异常聚集的影响。方法90只健康雄性 C57BL/6小鼠，鼠龄8~12w。随机分为3组，正常对照组30只、PD 模型对照组30只、PD 模型干预组30只。MPTP 腹腔注射40mg·Kg-1·d-1×7制备小鼠 PD 模型；正常对照组及 PD 模型对照组予生理盐水1ml·d-1灌胃；PD 模型干预组给予“抗帕颗粒”40mg·kg-1·d-1灌胃，连续喂养4个月。比较分析各组4月时黑质纹状体区α-syn聚集情况。结果①正常对照组30只（30/30只）最终均存活，PD 模型对照组4个月时存活27只（27/30只）， PD 模型干预组4个月时存活28只（28/30只）；② PD 模型对照组、PD 模型干预组小鼠每次注射 MPTP 后，先有短暂兴奋（持续7.61±2.17min），表现为四处窜跳；随即出现全身中重度震颤，皮毛及尾巴时有竖立，活动减少，持续24.23±3.89min 后震颤消失；随后出现活动减少；③黑质纹状体区α-syn 免疫荧光双染检测；③ PD 模型对照组较正常对照组明显增多，PD 模型干预组较 PD 模型对照组有所减少，但仍较正常对照组增多；④ Western blotting 检测α-syn 蛋白量的表达，正常对照组与 PD 模型对照组比较，P<0.001；与PD 模型干预组比较，P<0.001；PD 模型对照组与 PD 模型干预组比较，P<0.05。结论“抗帕颗粒”对 PD小鼠黑质纹状体区α-syn 异常聚集具有一定程度的抑制作用，阻断氧化应激反应可能是这种保护作用的关键机制，有望改善 PD 治疗现状并在临床进一步推广应用。%Objective To explore the effect of "anti-Parkinson granule" on synaptic nigrostriatal area á-synuclein(á-syn)anomaly aggregation in methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)Parkinson's disease(PD)model mice. Method Randomly divided 90 healthy male C57BL/6mice, with ages arrange 8-12 weeks, into 3 groups including of the normal control group, the PD model control group and the PD model intervention group, each have 30 mice. The PD model group were made through intraperitoneal injection with MPTP(40mg·kg-1·d-1×7). The normal control group and the PD model control group were pumped stomach with normal saline(1ml·d-1); The PD model intervention group were pumped stomach with anti-Parkinson granule solution(40mg·kg-1·d-1), and were fed continually for 4 months . At the end of the forth month, contrasted and analysised each group's status on accumulation of α-syn. Results ① 30 mice(30/30 mice)were survival in the normal control group; the PD model control group at end of 4 months survived 27 mice(27/30 mice); and the PD model intervention group at end of 4 months survived 28 mice(28/30 mice). ② After injection with MPTP, mice of the PD model control group and the PD model intervention group had short excitements(duration 7.61±2.17 min), behaved jumping around, followed with the whole body's moderate or severe tremors, fur and tails erected timely, less movement. Tremors ceased after 24.23±3.89 min, and then movement became less. ③ α-syn double immunofluorescence staining in Nigrostriatal area of PD model group increased significantly compared with that of normal control group, α-syn of the intervention group decreased compared with that of the PD model group, but still showed an increase compared with that of the control group. ④ Western blotting was used to detect the expression of α-syn protein. The results showed that there were significantly difference between the normal control group and PD model group(P<0.001), also with PD model intervention group P<0.001, there were significantly difference between PD model group and PD intervention model group(P<0.05). Conclusion " anti-Parkinson granule " could inhibit in certain degree the abnormal α-syn aggregation on PD mouse nigrostriatal area. The key mechanism of this protective effect may be through blocking the oxidative stress. It would be applicable to improve the situation of PD treatment in further clinical application.