基于铁死亡探究Nrf2/HO-1信号通路对帕金森病模型小鼠行为学障碍的调控作用

Regulating effects of Nrf2/HO-1 signal pathway on ferroptosis-induced behavioral dysfunction in mouse model of Parkinson's disease

目的探究核因子E2相关因子2/血红素加氧酶-1(Nuclear factorNF-E2-related factor 2/heme oxygenase,Nrf2/HO-1)信号通路对帕金森病(Parkinson's disease,PD)模型小鼠行为学障碍的改善作用及其调控机制。方法制备小鼠1-甲基-4-苯基-1,2,3,6-四氢吡啶(1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine,MPTP)模型,并给予Nrf2抑制剂ML385或铁死亡抑制剂Ferrostatin-1;实验结束后评估各组小鼠行为学表型、多巴胺能神经元损伤程度、氧化应激水平、Nrf2/HO-1信号通路和铁死亡指标表达水平。结果在MPTP小鼠模型中ML385加剧MPTP组小鼠行为学障碍及酪氨酸羟化酶(Tyrosine hydroxylase,TH)阳性神经元丢失,进一步诱导Fe^(2+)沉积、活性氧(Reactive oxygen species,ROS)合成及丙二醛(Malondialdehyde,MDA)分泌,降低超氧化物歧化酶(Superoxide dismutase,SOD)和谷胱甘肽(Glutathione,GSH)的水平,增加酰基辅酶A合成酶长链家族成员4(Acyl coenzyme a synthetase long chain family member 4,ACSL4)的蛋白表达,并抑制溶质载体家族7成员11(Solute carrier family 7 member 11,SLC7A11)和谷胱甘肽过氧化酶4(Glutathione peroxidase 4,GPX4)的蛋白表达;给予Ferrostatin-1后可取消上述ML385介导的神经损伤作用,同时不影响Nrf2和HO-1的蛋白表达水平。结论Nrf2/HO-1信号通路能够改善PD模型小鼠行为学障碍,其机制可能与抑制神经元铁死亡有关。

Objective The purpose of this study is to investigate the ameliorative effects and the regulatory mechanisms of Nrf2/HO-1 signal pathway on behavioral dysfunction in mouse model of Parkinson's disease(PD).Methods Made preparation of MPTP mice model.In the MPTP mice model,ML385(Nrf2 inhibitor)or Ferrostatin-1(ferroptosis inhibitor)was administered.Behavioral analysis,dopaminergic neuronal losses,oxidative stress,and the expression of the proteins involved in the Nrf2/HO-1 signaling pathway and ferroptosis were evaluated after the experimental manipulation in mice.Results ML385 exacerbated behavioral deficits,tyrosine hydroxylase(TH)-positive neuronal losses,also further led to the overloaded iron deposition,ROS accumulation and Malondialdehyde(MDA)excretion.Meanwhile,ML385 decreased the level of SOD and glutathione(GSH),increased the protein expression of ACSL4 and the suppressed the protein expression of SLC7A11 and GPX4 in mouse model of PD.By contrast,administration of Ferrostatin-1 abolished the above ML385-mediated neurological damage effects without affecting the protein expression levels of Nrf2 and HO-1.Conclusion Nrf2/HO-1 signal pathway can improve behavioral impairment in MPTP-induced mice,and perhaps its mechanism is related to inhibiting ferroptosis of dopaminergic neuron.