摘要
目的探究MPTP诱导C57BL/6小鼠亚急性帕金森(PD)模型脑TrkA和TrkB酪氨酸激酶阳性表达的情况。方法将C57BL/6小鼠随机分为对照(CON)组和MPTP 5 d组,腹腔注射MPTP制备C57BL/6小鼠亚急性PD模型。通过爬杆实验观察分析各组小鼠的运动行为学改变。应用免疫荧光染色、Western blot测定PD模型前后各组小鼠中脑黑质TrkA和TrkB的阳性表达情况。结果爬杆实验结果显示,与CON组相比,MPTP 5 d组小鼠爬杆运动时间减少(P<0.001)。荧光染色结果显示,中脑黑质上神经胶质酸性蛋白(GFAP)主要在星形胶质细胞胞浆和突起上表达,MPTP 5 d组中脑黑质腹侧、大脑脚及其周围的GFAP阳性细胞数增加;DA神经元的特异性标记物TH主要在神经元胞浆和突起上表达,MPTP 5 d组黑质致密部TH阳性细胞数减少;TrkA在DA神经元或星形胶质细胞胞浆上表达,MPTP 5 d组中脑黑质致密部和网状部TH和TrkA共标记的阳性细胞数减少,但纹状体区、中脑黑质腹侧以及大脑脚周围TrkA阳性细胞数增加;TrkB主要在DA神经元上表达,MPTP 5 d组中脑黑质致密部及其周围TH和TrkB共标记的阳性细胞数减少,且黑质致密部周围TrkB阳性细胞数减少。Western blot结果显示,与CON组相比,MPTP 5 d组中脑黑质TrkA和TrkB蛋白表达量减少(P均<0.001)。结论亚急性PD模型小鼠运动学习、活动能力下降,中脑黑质星形胶质细胞异常增生,且DA神经元TH、TrkA和TrkB阳性细胞数明显减少,TrkA和TrkB蛋白表达水平明显下调。
Objective To explore the changes of tyrosine kinases TrkA and TrkB positive expression in MPTP-induced PD model of C57BL/6 mice.Methods The PD mouse model was successfully induced by intraperitoneal injection of MPTP and were randomly divided into a control(CON)group and an MPTP 5 d group.The behavioral changes in each group were analyzed by the pole test.The positive expression of TrkA and TrkB in the substantia nigra of each group was determined by immunofluorescence staining and western blot protein analysis.Results The pole test results showed that the moving time in MPTP group had a significantly reduced(P<0.001 vs CON).By fluorescence staining the results indicated GFAP was mainly expressed in the cytoplasm and processes of astrocytes in the substantia nigra of the midbrain.The number of GFAP-positive cells in the lateral substantia nigra,crus cerebri and their surroundings in the MPTP 5 d group significantly increased compared with that of CON group.TH(a specific marker of DA neurons)was mainly expressed in the cytoplasm and processes of neurons.Compared with the CON group,the number of TH-positive cells in the substantia nigra pars compacta in the MPTP 5d group significantly decreased.TrkA was expressed in the cytoplasm of neurons or astrocytes.Compared with the CON group,the number of TH and TrkA co-labeled positive cells in the substantia nigra pars compacta of the MPTP 5d group was significantly reduced,but the number of TrkA positive cells in the lateral substantia nigra,crus cerebri and their surroundings was significantly increased.TrkB was expressed in the cytoplasm of neurons or astrocytes.Compared with the CON group,the number of TH and TrkB co-labeled positive cells in the substantia nigra pars compacta of the MPTP 5d group was significantly reduced,and the number of TrkB positive cells around the substantia nigra pars compacta was also significantly decreased.By Western blot protein analysis the results showed that the expression levels of TrkA and TrkB proteins in the MPTP 5d group were decreased,and there was a statistically difference compared with the CON group(P all<0.001).Conclusion The MPTP-induced PD model in C57BL/6 mice may lead to a decline in motor learning and activity ability.Moreover,in the PD model there is an abnormal proliferation of astrocytes,a significant decrease in the number of TH,TrkA and TrkB positive cells in DA neurons,and a significant down-regulation of TrkA and TrkB protein expression in the substantia nigra of the midbrain.
作者
丁银秀
刘丝雨
严青
王梦
靳国华
焦旭文
DING Yinxiu;LIU Siyu;YAN Qing;WANG Meng;JIN Guohua;JIAO Xuwen(Department of Human Anatomy,School of Basic Medical Sciences,Ningxia Medical University,Yinchuan 750004,China)
出处
《宁夏医科大学学报》
2025年第9期883-888,共6页
Journal of Ningxia Medical University
基金
宁夏自然科学基金项目(2023AAC03229)。
关键词
酪氨酸激酶Trks信号通路
中脑黑质
星形胶质细胞异常增生
帕金森病
tyrosine kinase Trks signaling pathway
midbrain substantia nigra
abnormal proliferation of astrocytes
Parkinson’s disease
