脑组织特异表达hS100B转基因小鼠帕金森样运动协调能力改变的初步分析  

Preliminary Analysis of Parkinson-like Motor Coordination Abnormity in Brain-specific hS100B Transgenic Mice

作　　者：刘嘉琳[1] 郑芳[2] 龙彦[2] 佟柳 郑媛[2] 刘小青[2] 秦川[3] 

机构地区：[1]河北大学医学部病理教研室,河北保定071000 [2]保定市第一中心医院老年病科,河北保定071000 [3]中国医学科学院医学实验动物研究所北京协和医学院比较医学中心,北京100021

出　　处：《中国医学科学院学报》2017年第2期240-246,共7页Acta Academiae Medicinae Sinicae

基　　金：保定市科学技术研究与发展指导计划(14ZF003);河北大学医学学科建设基金(2014A1002);河北大学博士科研启动基金(一省一校专项经费)~~

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。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.

关 键 词：S100B 帕金森病 多巴胺受体 G蛋白偶联受体激酶 多巴胺 

分 类 号：R741[医药卫生—神经病学与精神病学]