小鼠肠道菌群失调与帕金森病发生发展的相关性研究  

Study on the correlation between gut dysbacteriosis and Parkinson's disease in mouse

作　　者：庞书涛 韩燕银 刘承伟[1] Pang Shutao;Han Yanyin;Liu Chengwei(Guilin Medical University,Guilin 541000;Linfen Vocational and Technical College,Linfen 041000,China)

机构地区：[1]桂林医学院,桂林541000 [2]临汾职业技术学院,临汾041000

出　　处：《神经解剖学杂志》2018年第4期433-440,共8页Chinese Journal of Neuroanatomy

基　　金：广西自然科学基金(2013GXNSFDA019018)

摘　　要：目的:探索肠道菌群失调与帕金森病的发生、发展的相关性,揭示帕金森病的发病起因。方法:雄性C57BL/6小鼠80只,随机分为A组(正常对照组:灌胃生理盐水和腹膜腔注射生理盐水)、B组(肠道菌群失调组:只灌胃头孢曲松CRO)、C组(仅腹膜腔注射MPTP)、D组(灌胃CRO+腹膜腔注射MPTP),每组20只。在肠道菌群失调基础上,长期小剂量给予MPTP制备帕金森病小鼠模型;进行爬杆、悬挂和滚轴实验通过组间对比评估运动行为学改变;通过分析盲肠指数、肠道菌落数和观察盲肠的组织学改变来确定肠道菌群失调;通过Western Blot及免疫荧光组织化学双染方法检测中脑黑质酪氨酸羟化酶(TH)、α-突触核蛋白的表达及寡聚化和纤维化程度。结果:悬挂实验、滚轴实验显示:C、D组在给药2周后,与A组相比均出现明显的运动行为学改变,随着给药时间的延长,这种趋势的改变更加明显,且具有统计学意义(P<0.01);其中D组比C组的改变更加明显,两者之间有显著性差异(P<0.01)。B组与正常对照A组相比,无显著性差异(P>0.05)。爬杆实验各组与A组相比均无明显差异(P﹥0.05)。Western Blot结果显示:C组、D组与同窝对照A组相比,TH的表达均出现了随给药时间推移而改变的明显降低趋势(P<0.01);且D组比C组的改变要早而显著(P<0.05);α-突触核蛋白未检测出。免疫荧光组织化学双标染色结果亦显示:肠道菌群失调的PD模型与同窝对照组相比,黑质TH阳性神经元的数目明显下降(P<0.01),但α-突触核蛋白未检测出。结论:肠道菌群失调可加剧MPTP诱导的帕金森病小鼠模型提前出现运动行为学改变,证明肠道菌群失调在帕金森病发生发展过程中扮演着重要角色。Objective：To reveal the correlation between the occurrence and development of Parkinson＇s disease（PD）and the intestinal dysbacteriosis and to explore the pathogenesis of PD.Methods：80 male C57 BL/6 mice were randomly divided into group A,group B,group C,group D and were 20 mice in each group.The A group mice of control group was received same of saline solution by the same way.The B group mice of intestinal dysbacteriosis group were only lavaged ceftriaxone（CRO）.The C group mice was received MPTP by the way of intraperitoneal injection.The D group mice were received by being both lavaged ceftriaxone and injected MPTP intraperitoneally.The PD model of intestinal flora imbalance was established by being given to ceftriaxone and MPTP by the way of long-term and lower doses.The behavior of motor function was assessed by the comparison of Pole test,Traction test and Rota-Rod test.The imbalance of intestinal flora was determined by analysis of cecal index and intestinal flora and observation of the histological changes of the cecum.The expression of tyrosine hydroxylase（TH） and alpha-synuclein and the order of oligomerization and fibrosiswere detected by the Western Blot（WB） and double dye immunofluorescence（DDI） method in the substantia nigra of mesencephalon.Results：In the Traction and Rota-Rod test,the C and D group mice was gradually showed significant movement impairment（P〈0.01） compared with control A group after 2 weeks.In particular,the D group was more significant difference than the C group（P〈0.01）.There was no significant difference between group A and B（P〈0.05）in the above tests.In the Pole test,there was no statistically significant difference between the mice in the experimental group and the control group（P〈0.05）.According to the result of WB,as time went on and the expression of TH from the C and D group mice was significantly lower in comparison with group A（P〈0.01）.Moreover,the change of group D was earlier than that of group C（P〈0.05）.Al

关 键 词：肠道菌群失调 帕金森病模型 酪氨酸羟化酶 Α-突触核蛋白 MPTP 小鼠 

分 类 号：R742.5[医药卫生—神经病学与精神病学]