SHH信号通路的激活对帕金森病模型小鼠的保护及其机制  

The protective effects and its mechnisms of purmorphamine on mice models of Parkinson's disease

作　　者：洪乐鹏[1] 汪光亮[1] 王迪[2] 王智明[1] 邵帅[1] 

机构地区：[1]广州医科大学人体解剖教研室,广州511436 [2]广州医科大学第二附属医院康复科,广州511436

出　　处：《神经解剖学杂志》2017年第5期586-592,共7页Chinese Journal of Neuroanatomy

基　　金：广东省医学科研基金项目(A2013239);广东省科技计划项目(201301)

摘　　要：目的:为探讨Sonic Hedgehog(Shh)信号通路激活剂,2,6,9-三取代嘌呤化合物(purmorphamine,PM)对帕金森病(Parkinson’s disease,PD)模型动物中小鼠身体的协调能力、多巴胺能神经元及小胶质细胞细胞活性的影响及可能机制。方法:将47只C57BL/6J雄性8周小鼠随机分为control组、PM组、MPTP组、PM+MPTP组,使用腹腔注射1-methyl-4-phenyl 1,2,3,6-tet-rahydropyridine(MPTP)制作小鼠的帕金森病模型。应用悬杆实验、免疫组织化学和实时荧光定量PCR(real-time quantitative PCR,Q-PCR)技术分别检测小鼠的运动协调能力,酪氨酸羟化酶(tyrosine hydroxylase,TH)阳性的多巴胺(DA)能神经元的形态、数目,Ionized calcium binding adapter molecule 1(Iba1)阳性的小胶质细胞的形态以及炎症因子TNF-α的表达情况。结果:(1)悬杆实验显示:在预处理PM 24 h后MPTP诱导的PD模型小鼠中,与MPTP组相比,PM+MPTP组小鼠后爪抓到横杆的时间明显缩短(P<0.01);而与control组相比,MPTP组中小鼠后爪抓到横杆的时间显著延长(P<0.01)。(2)TH免疫组织化学结果显示:control组TH+DA能神经元染色很深,胞体密集,突起明显;与control组相比,MPTP组TH+DA能神经元的数量明显减少,细胞质着色变浅,突起变短或者消失(P<0.01);与MPTP组相比,PM+MPTP组TH+DA能神经元不仅使损失细胞的数目明显减少,而且细胞质染色变深,突起有所增多(P<0.01)。(3)Iba1免疫组织化学染色显示:与control相比,MPTP组小胶质细胞变大、形态不规则、突起变粗;与MPTP组相比,PM+MPTP组小胶质细的胞体形态较小,突起呈现细长较多。(4)Q-PCR检测结果显示:与control组相比,MPTP组TNF-α和Iba1mRNA表达明显增加(P<0.01);与MPTP组对比,PM+MPTP组表达的TNF-α和Iba1 mRNA水平明显降低(P<0.01)。结论:PM可改善PD模型小鼠的运动协调能力,保护多巴胺能神经元,其机制可能与降低小胶质细胞的炎症水平有关。Objective： To testify the effect of Sonic Hedgehog （Shh） signaling pathway activator Purmorphamine （PM） on mice model of Parkinson＇s disease（PD） and the changes of coordination, dopamine neurons and microglia cell activity. Methods： Forty-seven 8-month old C57BL/6J male mice were randomly divided into 4 groups, i. e. control group, PM group, MPTP group, PM ＋ MPTP group. The animal models of Parkinson＇s disease was built by intraperitone- al injection of 1 - methyl 4-phenyl 1,2,3,6 - tet-rahydropyridine （MPTP）. Bar test, immunohistochemistry and Real- time quantitative PCR （Q-PCR） technology were used to detect the motor coordination ability of mice, the shape and number of tyrosine hydroxylase（TH）-positive dopaminergic（DA） neurons, the shape of Ionized calcium binding adapter molecule 1 （ Ibal ） -positive microglia and the expression of inflammatory factor TNF-α . Results： （ 1 ） Bar test show that after pretreatment with PM for 24 h in the MPTP induced PD model mice, there was a significantly decreased crawl time in the PM ＋ MPTP group, compared with the MPTP group （P 〈 0.01 ） ; however, there was a significantly increased crawl time in the MPTP group vs the control group （P 〈0.01 ）. （2） TH immunohistochemistry staining showed： the TH＋ DA neurons in the control group were intensely stained, with compressed cell body and the processes were clear to see; com- pared with the control group, the neurons in MPTP group had a significantly decreased number, shallow stained, the processes were shorter or disappeared; there were a significantly increased number of TH＋ DA neurons in PM ＋ MPTP group, intensely stained and the processes were more, compared with the MPTP group（P 〈 0.01 ）. （3） Ibal immunohis- tochemistry staining showed ： the microglia in MPTP group became large, the shape of the cell body was irregular and the processes were thicker, compared with control group; the microglia in PM ＋ MPTP group was smaller, the processes w

关 键 词：Purmorphamine 帕金森病 炎症 小胶质细胞 小鼠 

分 类 号：R-332[医药卫生] R742.5