Li Jie, Xu Sui
Department of Clinical Medicine, Suzhou Vocational Health College, Suzhou, Jiangsu 215009, P.R. China.
Department of Traditional Chinese Medicine, Huamu Community Health Service Center, Shanghai 200120, P.R. China.
Exp Ther Med. 2022 Apr;23(4):293. doi: 10.3892/etm.2022.11223. Epub 2022 Feb 17.
The flavonoid tilianin is derived from the leaves of L. amiales and has been proven to serve a neuroprotective role in cerebral ischemia. Therefore, the aim of the present study was to determine whether tilianin could prevent oxidative stress and the apoptosis of dopaminergic neurons in Parkinson's disease (PD). The dopaminergic neuron MES23.5 cell line was treated with 1-methyl-4-phenylpyridinium (MPP) to construct a PD cell model. Following pretreatment with tilianin, the Cell Counting Kit-8 assay was used to assess cell viability. The protein and mRNA expression levels of tyrosine hydroxylase were determined using immunofluorescence, reverse transcription-quantitative PCR (RT-qPCR) and western blotting. mRNA and protein expression levels of inflammatory cytokines IL-6, IL-1β and TNF-α and oxidative stress-related enzymes manganese superoxide dismutase and catalase were also quantified using RT-qPCR and western blotting, respectively. Cell apoptotic rate was analyzed using the TUNEL assay and the expressions of apoptosis-related proteins Bcl-2, Bax and cleaved caspase-3 were detected by western blotting. MAPK signaling pathway-related protein expression levels were assessed via western blotting in MPP-stimulated MES23.5 cells with or without tilianin pretreatment. Tilianin was demonstrated to exert no cytotoxic effects on MES23.5 cells and was able to prevent MPP-induced reductions in cell viability. Pretreatment with tilianin also inhibited MPP-induced inflammatory cytokine secretion, oxidative stress and apoptosis of MES23.5 cells. In addition, the protein expression levels of MAPK signaling pathway-related proteins were upregulated by MPP, whereas pretreatment with tilianin downregulated these in a dose-dependent manner. The results of the present study indicated that tilianin may exert anti-inflammatory and antioxidant effects and inhibit the MAPK signaling pathway, which may ameliorate injury to dopaminergic neurons induced by PD.
黄酮类化合物田七素源自荔枝草的叶子，并且已被证明在脑缺血中发挥神经保护作用。因此，本研究的目的是确定田七素是否可以预防帕金森病（PD）中多巴胺能神经元的氧化应激和凋亡。用1-甲基-4-苯基吡啶鎓（MPP）处理多巴胺能神经元MES23.5细胞系以构建PD细胞模型。用田七素预处理后，使用细胞计数试剂盒-8法评估细胞活力。使用免疫荧光、逆转录定量PCR（RT-qPCR）和蛋白质印迹法测定酪氨酸羟化酶的蛋白质和mRNA表达水平。还分别使用RT-qPCR和蛋白质印迹法定量炎性细胞因子IL-6、IL-1β和TNF-α以及氧化应激相关酶锰超氧化物歧化酶和过氧化氢酶的mRNA和蛋白质表达水平。使用TUNEL法分析细胞凋亡率，并通过蛋白质印迹法检测凋亡相关蛋白Bcl-2、Bax和裂解的caspase-3的表达。通过蛋白质印迹法评估在有或没有田七素预处理的MPP刺激的MES23.5细胞中MAPK信号通路相关蛋白的表达水平。结果表明，田七素对MES23.5细胞无细胞毒性作用，并且能够预防MPP诱导的细胞活力降低。田七素预处理还抑制了MPP诱导的MES23.5细胞炎性细胞因子分泌、氧化应激和凋亡。此外，MPP上调了MAPK信号通路相关蛋白的蛋白质表达水平，而田七素预处理以剂量依赖性方式下调了这些蛋白的表达水平。本研究结果表明，田七素可能发挥抗炎和抗氧化作用，并抑制MAPK信号通路，这可能改善PD诱导的多巴胺能神经元损伤。
