利用七氯对斑马鱼多巴胺神经元毒性影响构建帕金森病模型  

Establishment of Parkinson’s disease model by the toxicity effect of heptachlor on dopamine neuron in zebrafish

作　　者：成慧灵 周美 许茜[2,3] 蔡晶 Cheng Huiling;Zhou Mei;Xu Qian;Cai Jing(Academy of Integrative Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;College of Integrated Chinese and Western Medicine,Fujian University of Traditional Chinese Medicine,Fuzhou 350122,China;Key Laboratory of Developmental and Neurobiology of Fujian Universities,School of Life Sciences,Fujian Normal University,Fuzhou 350117,China)

机构地区：[1]福建中医药大学中西医结合研究院,福州350122 [2]福建中医药大学中西医结合学院,福州350122 [3]福建师范大学生命科学学院发育与神经生物学福建省高校重点实验室,福州350117

出　　处：《广西医科大学学报》2022年第7期1047-1053,共7页Journal of Guangxi Medical University

基　　金：国家自然科学基金青年项目(No.81904263);2021-2024年度福建省中医药科研项目计划(No.2021zyjc02)。

摘　　要：目的:利用七氯对斑马鱼多巴胺神经元的影响,建立斑马鱼帕金森病(PD)模型。方法:将斑马鱼鱼卵分为正常对照组(不予干预),750μmol/L 6-羟基多巴胺(6-OHDA)组和不同浓度七氯组(1 ng/mL、10 ng/mL、100 ng/mL、1000 ng/mL、10000 ng/mL),分别恒温孵育,每24 h观察胚胎发育情况并记录直到受精后120 hpf(h post fertilization,hpf),在120 hpf做行为学测定,然后实时荧光定量PCR(RT-qPCR)测定多巴胺通路酪氨酸羟化酶(TH2)的相对表达量。结果:与正常对照组比较,6-OHDA组幼体出现发育迟缓、脊柱弯曲,运动距离缩短(P<0.05),TH2基因表达降低(P<0.05);与正常对照组比较,1~100 ng/mL七氯组幼体发育无明显变化,但运动距离缩短(P<0.05),TH2基因表达降低(P<0.05),1000~10000 ng/mL七氯组出现色素沉着分布和脊柱弯曲,而且运动距离更短(P<0.05)、TH2基因表达更低(P<0.05),变化具有浓度依赖性;与6-OHDA组比较,1000~10000 ng/mL七氯组运动距离更短,TH2基因表达更低,其中10000 ng/mL七氯组运动距离缩短和TH2基因表达降低有统计学差异(P<0.05)。结论:传统PD模型造模药物6-OHDA和1000~10000 ng/mL七氯都可通过影响多巴胺神经元功能成功建立斑马鱼PD模型,其中10000 ng/mL七氯的造模效果比6-OHDA更显著。Objective:To establish the model of Parkinson’s disease(PD)by using the effect of heptachlor on dopamine neuronal toxicity in zebrafish.Methods:Zebrafish eggs were divided into normal control group(without intervention),750μmol/L 6-hydroxydopamine(6-OHDA)group and different concentrations of heptachlor group(1 ng/mL,10 ng/mL,100 ng/mL,1,000 ng/mL,10,000 ng/mL).The embryonic development was observed every 24 h and recorded until 120 h post fertilization(hpf)after fertilization.Then the relative expression of tyrosine hydroxylase(TH2)in dopamine pathway was measured by real-time quantitative PCR.Results:Compared with the normal control group,the larvae of 6-OHDA group showed developmental retardation,rachiocamposis,shorter distance of movement(P<0.05)and lower expression of TH2 gene(P<0.05).Compared with the normal control group,the larval development of 1-100 ng/mL heptachlor groups had no significant change,but the movement distance was shorter(P<0.05)and the expression of TH2 gene was decreased(P<0.05).1,000 and 10,000 ng/mL heptachlor groups showed pigmentation distribution and rachiocamposis,and the movement distance was shorter(P<0.05)and the expression of TH2 gene was lower(P<0.05),showing concentration dependent.Compared with 6-OHDA group,1,000 and 10,000 ng/mL heptachlor group had shorter exercise distance and lower TH2 gene expression.Among them,10,000 ng/mL heptachlor groups had shorter exercise distance and significant lower TH2 gene expression(P<0.05).Conclusions:Both traditional PD modeling drugs 6-OHDA,1,000 and 10,000 ng/mL heptachlor can successfully establish zebrafish PD model by affecting the function of dopamine neurons,and the modeling effect of 10,000 ng/mL heptachlor is more significant than that of 6-OHDA.

关 键 词：七氯 多巴胺神经元 斑马鱼 帕金森病 模型 

分 类 号：R742.5[医药卫生—神经病学与精神病学]