小檗碱衍生物9-OH小檗碱的抗帕金森病活性和作用机制研究

Study on the activity and mechanism of berberine derivative 9-OH berberine against Parkinson’s disease

目的:探究小檗碱衍生物9-OH小檗碱对1-甲基-4-苯基-1,2,3,6-四氢吡啶盐酸盐(1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride,MPTP)诱导的斑马鱼帕金森病(Parkinson’s disease,PD)症状是否具有缓解作用。方法:受精后1 d(day post fertilization,dpf)的斑马鱼胚胎,设置空白对照组,50μmol·L^(-1)MPTP造模组,50μmol·L^(-1)MPTP与不同质量浓度(100,300,500μmol·L^(-1))9-OH小檗碱共处理组。4 dpf时,分别观察并记录各组斑马鱼多巴胺神经元及脑部血管的发育情况;5 dpf时,监测行为学,记录各组行为学数据并分析差异,收集后检测PD相关基因表达变化,包括编码α-突触核蛋白(α-synuclein,α-Syn)的α-syn、编码E3泛素连接酶(E3 ubiquitin protein ligase,Parkin)的parkin、编码自噬相关基因5(autophagy related gene 5,Atg5)的atg5、编码微管相关蛋白1轻链3B(microtubule-associated protein 1 light chain 3B,Lc3b)的lc3b。结果:MPTP造模组的斑马鱼多巴胺神经元和脑部血管出现不同程度的损伤,且出现运动迟缓、焦虑等PD状行为,与PD相关的基因表达也出现异常;而MPTP与9-OH小檗碱共处理组的斑马鱼多巴胺神经元和脑部血管的损伤得到改善,行为学能力得到恢复,且PD相关基因的表达趋于正常。结论:小檗碱衍生物9-OH小檗碱对MPTP诱导的斑马鱼帕金森病症状有缓解作用。

OBJECTIVE To investigate whether berberine derivative 9-OH berberine can relieve symptoms of Parkinson’s disease(PD)induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine hydrochloride(MPTP)in zebrafish.METHODS Zebrafish embryos with one day post fertilization(dpf)were demembraned with release agent,and then divided into different groups including control group,50μmol·L^(-1)MPTP model group,and 50μmol·L^(-1)MPTP with different concentrations(100,300,500μmol·L^(-1))of 9-OH berberine groups.At 4 dpf,the development of dopamine neurons and cerebral blood vessels in each group was observed and recorded.At 5 dpf,behavioral data were monitored,behavioral data of each group were recorded and the differences were analyzed,and PD-related gene expression changes were detected after collectio,and the genes includedα-synuclein(α-syn),E3 ubiquitin protein ligase(parkin),autophagy related gene 5(atg5)and microtubule-associated protein 1 light chain 3 B(lc3 b).RESULTS In the MPTP model group,dopamine neurons and brain blood vessels were damaged to varying degrees,and PD-like behaviors such as bradykinesia and anxiety were found,and PD-related gene expression was also abnormal.However,the damage of dopamine neurons and cerebral blood vessels in zebrafish co-treated with MPTP and 9-OH berberine was improved,the behavioral ability was recovered,and the expression of PD-related genes tended to be normal.CONCLUSION Berberine derivative 9-OH berberine shows ameliorative effect on MPTP-induced Parkinson’s symptoms in zebrafish.