背景:灵芝,一种著名的补品中药,在舒缓神经和滋养大脑方面的特殊活动得到了广泛认可。它已被广泛用于缓解各种神经系统疾病,尤其是帕金森病(PD)。
目的:评价GAA的抗帕金森病作用,灵芝的主要生物活性成分,并通过铁细胞自噬介导多巴胺能神经元铁凋亡的角度阐明其分子机制。
方法:使用1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)和1-甲基-4-苯基吡啶(MPP)建立PD小鼠和细胞模型,分别。细胞活力,进行行为测试和免疫荧光分析以评估神经毒性,运动功能障碍和多巴胺能丧失,分别。生化检测试剂盒用于测定铁的水平,脂质活性氧(ROS),丙二醛(MDA),总ROS和谷胱甘肽(GSH)。免疫印迹法和免疫荧光法检测核受体共激活因子4(NCOA4)的表达,铁蛋白重链1(FTH1),P62和LC3B。此外,构建过表达NCOA4的质粒载体,以验证GAA对PD模型中神经毒性和铁凋亡相关参数的抑制作用。
结果:GAA显著减轻MPP+/MPTP诱导的神经毒性,运动功能障碍和多巴胺能神经元丢失(p<0.01或p<0.05)。与MPP+/MPTP治疗相反,GAA治疗降低了铁的水平,MDA,脂质和总ROS,同时提高GSH水平。GAA还降低了NCOA4和LC3B的水平,并增强PD模型中FTH1和p62的表达(p<0.01或p<0.05)。然而,GAA对神经毒性的保护作用,NCOA4的过表达消除了PD模型中NCOA4介导的铁吞噬和铁凋亡(p<0.01)。
结论:GAA对PD有保护作用,这种作用是通过抑制NCOA4介导的铁细胞吞噬来抑制多巴胺能神经元铁凋亡来实现的。
BACKGROUND: Ganoderma lucidum, a renowned tonic traditional Chinese medicine, is widely recognized for the exceptional activity in soothing nerves and nourishing the brain. It has been extensively employed to alleviate various neurological disorders, notably Parkinson\'s disease (PD).
OBJECTIVE: To appraise the antiparkinsonian effect of GAA, the main bioactive constituent of G. lucidum, and clarify the molecular mechanism through the perspective of ferritinophagy-mediated dopaminergic neuron ferroptosis.
METHODS: PD mouse and cell models were established using 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) and 1-methyl-4-phenylpyridinium (MPP+), respectively. Cell viability, behavioral tests and immunofluorescence analysis were performed to evaluate the neurotoxicity, motor dysfunction and dopaminergic loss, respectively. Biochemical assay kits were used to determine the levels of iron, lipid reactive oxygen species (ROS), malondialdehyde (MDA), total ROS and glutathione (GSH). Western blot and immunofluorescence were applied to detect the expressions of nuclear receptor co-activator 4 (NCOA4), ferritin heavy chain 1 (FTH1), p62 and LC3B. Additionally, NCOA4-overexpressing plasmid vector was constructed to verify the inhibitory effect of GAA on the neurotoxicity and ferroptosis-related parameters in PD models.
RESULTS: GAA significantly mitigated MPP+/MPTP-induced neurotoxicity, motor dysfunction and dopaminergic neuron loss (p<0.01 or p<0.05). In contrast to MPP+/MPTP treatment, GAA treatment decreased the levels of iron, MDA, lipid and total ROS, while increasing the GSH level. GAA also reduced the levels of NCOA4 and LC3B, and enhanced the expressions of FTH1 and p62 in PD models (p<0.01 or p<0.05). However, the protective effect of GAA against the neurotoxicity, NCOA4-mediated ferritinophagy and ferroptosis in PD model was abolished by the overexpression of NCOA4 (p<0.01).
CONCLUSIONS: GAA exerted a protective effect on PD, and this effect was achieved by suppressing dopaminergic neuron ferroptosis through the inhibition of NCOA4-mediated ferritinophagy.