【摘要】 目的研究S100B在帕金森病发病机制中的作用并探讨脑组织特异表达hS100B转基因小鼠作为帕金森病模型小鼠的可能性。方法构建hS100B转基因小鼠。小鼠分为S100B转基因组、基因敲除组和阴性对照组。爬杆法和转棒法检测运动协调能力;逆转录-PCR、Western blot对脑组织中S100B、多巴胺D1、D2受体,G蛋白偶联受体激酶2、5及酪氨酸羟化酶的表达进行测定;高效液相色谱-荧光法检测脑组织中酪氨酸、左旋多巴、多巴胺及高香草酸的含量。结果与阴性对照组相比,转基因组小鼠脑组织中S100B蛋白显著升高(P<0.05),3月龄时可表现出运动协调能力下降,且随着月龄增加,运动协调能力下降呈明显的进行性降低(P<0.05);脑组织中多巴胺D2受体、G蛋白偶联受体激酶2mRNA和蛋白表达降低(P<0.05);左旋多巴、多巴胺、高香草酸生成增多,高香草酸/多巴胺比值升高(P<0.05);酪氨酸羟化酶的表达虽呈下降趋势,但差异无统计学意义(P>0.05)。S100B基因敲除小鼠脑组织中无S100B蛋白表达,与阴性对照组相比,各检测指标无异常(P>0.05)。结论 S100B很可能是参与帕金森病行为学异常的重要蛋白,脑组织特异表达hS100B转基因小鼠的建立为研究该基因在帕金森病中的作用提供了工具。更多还原

【Abstract】 Objective To investigate the role of S100B in the development of Parkinson ’s disease( PD) and explore the possibility of brain-specific S100B transgenic mice as PD animal model. Methods The hS100B transgenic mice were established. The mice were divided into S100B transgenic group( TG),S100B knockout group( KG),and the non-transgenic control group( CG). Motor coordination ability of mice was measured by the rota-rod and pole-climbing test. The expressions of S100 B,dopamine D1 receptor,dopamine D2 receptor,G protein-coupled receptor kinase( GRK) 2,GRK5,and tyrosine hydroxylase in brain tissue were detected by reverse transcription-polymerase chain reaction and Western blot. The levels of tyrosine,levodopa,dopamine,and homovanillic acid in brain tissue were measured by high-performance liquid chromatography coupled with fluorescence detection. Results Compared with CG,the S100B protein expression in brain tissue significantly increased in TG( P < 0. 05); the motor coordination ability of mice showed progressive decline( P < 0. 05); the mRNA and protein expressions of dopamine D2 receptor and GRK2 significantly decreased( P < 0. 05); the levels of levodopa,dopamine,and homovanillic acid were significantly elevated( P < 0. 05);the expression of tyrosine hydroxylase was also down-regulated,although there was no significant difference( P >0. 05). Compared with CG,there was no obvious change of the above indicators in KG( all P > 0. 05).Conclusions S100B plays an important role in the motor coordination abnormity of PD. The brain-specific S100B transgenic mice can be used in research on the role of S100B gene in the development of PD.更多还原