摘要
【目的】通过运动症状表现和脑组织病理变化指标评价皮下和肌内注射1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)构建的树鼩帕金森病(Parkinson’s disease,PD)模型,为PD研究提供理想的树鼩模型。【方法】通过皮下和肌内注射MPTP药液2 mg/kg诱导树鼩PD模型,参考非人灵长类和PD患者运动症状量化评估标准,从震颤、姿势、运动、步态及平衡5个方面建立树鼩PD模型行为评分表,观察树鼩PD运动症状;通过酪氨酸羟化酶(tyrosine hydroxylase,TH)和α-突触核蛋白(α-synuclein,α-syn)免疫组织化学染色观察树鼩中脑形态学的变化;通过qPCR检测分析树鼩PD模型脑组织中TH和α-syn的mRNA表达。【结果】2种注射方法均在第3次注射后出现典型的PD运动症状,第4次注射后PD运动症状最明显,第7次注射后PD运动症状明显减弱,第15次注射时基本消失;TH和α-syn免疫组织化学染色检测发现:皮下和肌内注射模型组黑质均有TH阳性神经纤维数量减少、α-syn异常增加的现象,均能模拟典型的PD病理学表现。皮下和肌内注射模型组脑组织TH mRNA表达量极显著降低,α-syn mRNA表达量极显著升高。【结论】重复皮下和肌内注射2 mg/kg MPTP药液均能诱导树鼩产生较一致的PD典型运动症状和病理变化,均是较好的树鼩PD模型造模方法。
[Purpose]To establish an ideal animal model of Parkinson’s disease(PD)in tree shrews,subcutaneous and intramuscular injections of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)were used to observe the motor symptoms and brain histopathological changes in the tree shrews.[Methods]The tree shrew PD model was induced by subcutaneous and intramuscular injections of 2 mg/kg MPTP,and the motor symptoms of PD were observed.A behavioral rating scale assessing tremor,posture,locomotion,gait,and balance was established for the tree shrew PD model with reference to the quantitative assessment criteria for Parkinson’s motor symptoms in non-human primates and patients;morphological changes in the midbrain of the tree shrews were observed by immunohistochemical staining of tyrosine hydroxylase(TH)andα-synuclein(α-syn).mRNA expression of TH andα-syn in the brain tissues of the tree shrew PD model was analyzed by qPCR.[Results]Typical motor symptoms of PD appeared after the 3rd injection in the two injection methods,and the most obvious PD motor symptoms were found after the 4th injection,which were obviously weakened after the 7th injection and basically disappeared in the 15th injection.Immunohistochemical staining of TH andα-syn revealed that there was a decrease in the number of TH-positive nerve fibers in the substantia nigra and an abnormal increase in the number ofα-syn in both the subcutaneous and intramuscular injections groups,demonstrating typical PD pathology.In the two injections groups,TH mRNA expression in the brain tissues was extremely significantly decreased andα-syn mRNA expression was extremely significantly increased.[Conclusion]Repeated subcutaneous and intramuscular injections of 2 mg/kg MPTP both can induce consistent motor symptoms and lesions typical of PD in tree shrews,and both can be used to model PD in tree shrews.
作者
李波
角建林
罗婷
冯所芹
孙怀昌
LI Bo;JIAO Jianlin;LUO Ting;FENG Suoqin;SUN Huaichang(Veterinary Medicine Department,Yangzhou University,Yangzhou 225009,China;Technology Transferring Center,Kunming Medical University,Kunming 650500,China)
出处
《云南农业大学学报（自然科学版）》
CAS
CSCD
北大核心
2024年第4期45-53,共9页
Journal of Yunnan Agricultural University:Natural Science
基金
云南省科技厅—昆明医科大学基础研究联合专项基金资助[2018FE001(-191),202401AY070001-064]。
作者简介
李波(1979-),女,云南保山人,在读博士,高级实验师,主要从事人类疾病动物模型构建与药物研究。E-mail:liboo@kmmu.edu.cn;通信作者:孙怀昌(1956-),男,江苏涟水人,博士,教授,主要从事兽医微生物学、免疫学和转基因制药研究。E-mail:sunh@yzu.edu.cn。
