牛磺酸对百草枯诱导的帕金森病样小鼠神经元及小胶质细胞的保护作用  

Protective effects of taurine on neurons and microglia in Parkinson's disease-like mouse model induced by paraquat

作　　者：田甜 张冰洋 王楷栋 张宝馥 黄敏 Tian Tian;Zhang Bingyang;Wang Kaidong;Zhang Baofu;Huang Min(Department of Occupational Health and Environmental Hygiene,School of Public Health and Management,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学公共卫生与管理学院职业卫生与环境卫生学系,银川750004

出　　处：《中华劳动卫生职业病杂志》2020年第11期801-808,共8页Chinese Journal of Industrial Hygiene and Occupational Diseases

基　　金：国家自然科学基金项目(81560538)。

摘　　要：目的探讨牛磺酸(Tau)对百草枯(PQ)诱导的帕金森病(PD)样小鼠海马、黑质区神经元及小胶质细胞的保护作用。方法于2019年4月,将36只SPF级C57BL/6小鼠按随机数字表法分为对照组(NaCl)、Tau对照组(150 mg/kg)、PQ染毒组(10 mg/kg PQ组、15 mg/kg PQ组)、Tau干预组(Tau+10 mg/kg PQ组、Tau+15 mg/kg PQ组),每组6只小鼠。腹腔注射PQ构建PD样小鼠模型并于PQ给药前1 h腹腔注射150mg/kg Tau进行干预,每周2次,连续6周。观察各组小鼠的一般和神经行为学(悬挂实验、旷场实验、游泳实验、悬尾实验、高架十字迷宫实验及新物体识别实验)变化,评估小鼠的运动能力和认知功能。染毒和干预结束后取小鼠脑组织进行尼氏染色检测海马、黑质区神经元尼氏体数量及形态变化;免疫组织化学染色(IHC)检测黑质区神经元标记物神经元核抗原(NeuN)、多巴胺(DA)能神经元标记物酪氨酸羟化酶(TH)、α-突触核蛋白(α-syn)、小胶质细胞标记物离子钙接头蛋白分子-1(Iba-1)、炎性因子诱导型一氧化氮合酶(iNOS)和白细胞介素-1β(IL-1β)的表达量;免疫荧光双标染色检测黑质区TH与Iba-1、TH与α-syn的共表达情况。结果 PQ染毒组小鼠出现反应迟钝、行动减少等一般行为学改变。与对照组比较,PQ染毒组小鼠悬挂实验得分、新物体识别时间比减少,游泳实验和悬尾实验不动时间增多(P<0.05);与对照组比较,15 mg/kg PQ组旷场实验水平爬格数、直立次数和高架十字迷宫实验开放臂停留时间比减少(P<0.05);与PQ染毒组比较,相同剂量Tau干预组小鼠悬挂得分增多,游泳和悬尾不动时间减少(P<0.05);与15 mg/kg PQ组比较,Tau+15 mg/kg PQ组旷场实验水平爬格数、新物体识别时间比增多(P<0.05)。尼氏染色和IHC结果显示,与对照组比较,PQ染毒组海马和黑质区的尼氏体数目减少(P<0.05),黑质区神经元NeuN、TH阳性细胞数目减少(P<0.05),α-syn和小胶质细胞Iba-1阳性表达量�Objective To investigate the protective effect of taurine(Tau)on hippocampus,substantia nigra neurons and microglia in paraquat(PQ)-induced Pakinson's disease-like mice.Methods In April 2019,the specific pathogen free(SPF)C57BL/6 mice(n=36)were randomly divided into control group(NaCl),Tau control group(150 mg/kg),PQ exposure group(10 mg/kg PQ group,15 mg/kg PQ group),Tau intervention group(Tau+10 mg/kg PQ group,Tau+15 mg/kg PQ group),respectively.Tau was used in 1 h before PQ administration for consecutive 6 weeks(twice per week).General and neurobehavioral tests(Traction test,Open field test,Forced Swimming test,Tail suspension test,High plus maze and Object recognition test)were performed to test motor and cognitive function.After neuroethology detection,mice were euthanized and brains were collected.Nissl staining was used to detect the changes of the number and morphology of Nissl bodies in hippocampus and substantia nigra neurons of mice.Immunohistochemistry(IHC)was used to test the levels of neuron marker neuronal nuclei antigen(NeuN),substantia nigra dopaminergic neuron marker tyrosine hydroxylase(TH),α-synuclein(α-syn),microglia markers ionized calcium bindingadaptor molecule-1(Iba-1),inducible nitric oxide synthase(iNOS)and interleukin-1β(IL-1β)in mice substantia nigra.The coexpression of Iba-1 and TH double-labeling,α-syn and TH double-labeling in mice substantia nigra were measured by immunofluorescence double staining.Results General behavioral changes such as slow reaction and reduced action occurred in mice of PQ group.Compared with the control group,the scores of Traction test,and the time ratio of new object recognition in the PQ group decreased(P<0.05),the fixed time of Swimming test and Tail suspension test increased(P<0.05),the horizontal crawl number and vertical times of Open field test and the ratio of open arm residence time of High plus maze in the 15 mg/kg PQ group decreased(P<0.05).Compared with the PQ group,the same dose of Tau+PQ group showed increased scores in Traction test(P<0

关 键 词：牛磺酸 百草枯 帕金森病 神经元 小胶质 小神经胶质细胞 小鼠 

分 类 号：R742.5[医药卫生—神经病学与精神病学]