Du Kechen, Su Ying, Song Qiong, Chen Shuai, Wu Ribao, Teng Xiahong, Huang Renbin, Wang Lihui, Zou Chunlin
Key Laboratory of Longevity and Aging-Related Disease of Chinese Ministry of Education, Center for Translational Medicine, School of Basic Medical Sciences, Guangxi Medical University, Nanning, Guangxi, China.
Center for Translational Medicine, School of Basic Medical Sciences, Guangxi Medical University, Nanning, Guangxi, 530021, China.
Metab Brain Dis. 2025 Jan 29;40(1):113. doi: 10.1007/s11011-025-01544-7.
2-dodecyl-6-methoxycyclohexa-2,5-diene-1,4-dione (DMDD) is a cyclohexanedione compound extracted from the roots of Averrhoa carambola L. Several studies have documented its beneficial effects on diabetes, Alzheimer's disease, and cancer. However, its potential neuroprotective effects on Parkinson's disease (PD) have not yet been explored. The present study aimed to investigate the protective effects and underlying mechanisms of DMDD in a cellular model of PD. In this study, SH-SY5Y cells were incubated with or without DMDD following intoxication with the parkinsonian neurotoxin 1-methyl-4-phenylpyridine (MPP). Cell viability and apoptosis were evaluated using 3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2 H-tetrazolium (MTS) assay and Hoechst 33,342 staining, respectively. The mitochondrial membrane potential (Δψm) was assessed through the JC-10 assay. The activities of superoxide dismutase (SOD) and the levels of reactive oxygen species (ROS) were measured using WST-8 and DCFH-DA assays. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed to explore significant biological processes and pathways influenced by DMDD. Molecular docking was employed to predict the domains of potential protein targets interacting with DMDD. Western blotting was subsequently conducted to determine the protein expression levels of TH, Nrf2, Bax, Bcl-2, Caspase-3, Beclin-1, PARP, LC3-II, LC3-I, p-PI3K, PI3K, p-mTOR and mTOR. Our study showed that DMDD treatment significantly increased cell viability and reduced apoptosis in MPP-treated SH-SY5Y cells. In addition, DMDD treatment reversed the loss of TH expression and Δψm in MPP-exposed SH-SY5Y cells. Moreover, DMDD treatment reduced MPP+-induced ROS production by promoting SOD activity. Additionally, compared with those in the MPP group, the protein expression levels of Beclin-1, Caspase-3, and PARP and the LC3II/I ratio were significantly decreased, whereas the protein expression levels of Nrf2 and the Bcl-2/Bax, p-PI3K/PI3K, and p-mTOR/mTOR ratios were significantly increased in the DMDD-treated group. In conclusion, DMDD protects against MPP-induced cytotoxicity by mitigating oxidative stress, apoptosis, and autophagy. PI3K/mTOR signaling at least partly mediates the cytoprotective effect of DMDD.
2-十二烷基-6-甲氧基环己-2,5-二烯-1,4-二酮（DMDD）是从杨桃根中提取的一种环己二酮化合物。多项研究记录了其对糖尿病、阿尔茨海默病和癌症的有益作用。然而，其对帕金森病（PD）的潜在神经保护作用尚未得到探索。本研究旨在探讨DMDD在PD细胞模型中的保护作用及潜在机制。在本研究中，用帕金森神经毒素1-甲基-4-苯基吡啶（MPP）处理SH-SY5Y细胞后，分别在有或无DMDD的情况下进行孵育。分别使用3-(4,5-二甲基噻唑-2-基)-5-(3-羧基甲氧基苯基)-2-(4-磺基苯基)-2H-四唑（MTS）法和Hoechst 33342染色评估细胞活力和凋亡。通过JC-10法评估线粒体膜电位（Δψm）。使用WST-8和DCFH-DA法测量超氧化物歧化酶（SOD）的活性和活性氧（ROS）水平。进行基因本体论（GO）和京都基因与基因组百科全书（KEGG）富集分析，以探索受DMDD影响的重要生物学过程和途径。采用分子对接预测与DMDD相互作用的潜在蛋白质靶点结构域。随后进行蛋白质免疫印迹法以确定TH、Nrf2、Bax、Bcl-2、Caspase-3、Beclin-1、PARP、LC3-II、LC3-I、p-PI3K、PI3K、p-mTOR和mTOR的蛋白质表达水平。我们的研究表明，DMDD处理显著提高了MPP处理的SH-SY5Y细胞活力并减少了凋亡。此外，DMDD处理逆转了MPP处理的SH-SY5Y细胞中TH表达的丧失和Δψm。此外，DMDD处理通过促进SOD活性降低了MPP+诱导的ROS产生。此外，与MPP组相比，DMDD处理组中Beclin-1、Caspase-3和PARP的蛋白质表达水平以及LC3II/I比值显著降低，而Nrf2的蛋白质表达水平以及Bcl-2/Bax、p-PI3K/PI3K和p-mTOR/mTOR比值显著升高。总之，DMDD通过减轻氧化应激、凋亡和自噬来保护细胞免受MPP诱导的细胞毒性。PI3K/mTOR信号通路至少部分介导了DMDD的细胞保护作用。
