Department of Neurology, The First Affiliated Hospital of Baotou Medical College, Inner Mongolia University of Science and Technology, 41 Linyin Road, Baotou, 014010, Inner Mongolia, China.
Neurochem Res. 2022 Oct;47(10):3137-3149. doi: 10.1007/s11064-022-03669-y. Epub 2022 Jul 9.
Miltirone is a phenanthrene-quinone derived from Salvia miltiorrhiza Bunge with anti-inflammatory and anti-oxidant effects. Our study aimed to explore the protective effect of miltirone on 1-methyl-4-phenylpyridinium (MPP)-induced cell model of Parkinson's disease (PD). PharmMapper database was employed to predict the targets of miltirone. PD-related genes were identified using GeneCards database. The overlapping genes between miltirone and PD were screened out using Venn diagram. KEGG analysis was performed using DAVID and KOBAS databases. Cell viability, reactive oxygen species (ROS) generation, apoptosis, and caspase-3 activity were detected by CCK-8 assay, a ROS assay kit, TUNEL, and caspase-3 activity assay, respectively. Effect of miltirone on the phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) pathway was explored by western blot analysis. A total of 214 targets of miltirone and 372 targets related to PD were attained, including 29 overlapping targets. KEGG analysis demonstrated that the 29 overlapping targets were both significantly enriched in the PI3K/Akt pathway. MPP stimulation reduced the cell viability in SH-SY5Y cells and neuronal primary cultures derived from human brain. Miltirone or N-acetylcysteine (NAC) attenuated MPP-induced reduction in cell viability, ROS production, SOD activity reduction, apoptosis, and increase of caspase-3 activity. Additionally, miltirone recuperated MPP-induced inactivation of the PI3K/Akt pathway. Moreover, treatment with LY294002, an inhibitor of the PI3K/Akt pathway, reversed the inhibitory effect of miltirone on MPP-induced ROS generation and apoptosis in SH-SY5Y cells and neuronal primary cultures. In conclusion, miltirone attenuated ROS-dependent apoptosis in MPP-induced cellular model of PD through activating the PI3K/Akt pathway.
丹参酮是一种来源于丹参的菲醌类化合物，具有抗炎和抗氧化作用。本研究旨在探讨丹参酮对 1-甲基-4-苯基吡啶离子（MPP）诱导的帕金森病（PD）细胞模型的保护作用。利用 PharmMapper 数据库预测丹参酮的作用靶点，利用 GeneCards 数据库鉴定 PD 相关基因，利用 Venn 图筛选出丹参酮与 PD 的重叠基因，利用 DAVID 和 KOBAS 数据库进行 KEGG 分析。通过 CCK-8 检测试剂盒、ROS 检测试剂盒、TUNEL 法和 caspase-3 活性检测试剂盒分别检测细胞活力、活性氧（ROS）生成、细胞凋亡和 caspase-3 活性。通过 Western blot 分析探讨丹参酮对磷脂酰肌醇 3-激酶（PI3K）/蛋白激酶 B（Akt）通路的影响。共获得丹参酮 214 个作用靶点和 372 个与 PD 相关的靶点，其中 29 个重叠靶点。KEGG 分析表明，这 29 个重叠靶点均显著富集在 PI3K/Akt 通路中。MPP 刺激降低了 SH-SY5Y 细胞和人脑中提取的原代神经元的细胞活力。丹参酮或 N-乙酰半胱氨酸（NAC）可减轻 MPP 诱导的细胞活力降低、ROS 生成增加、SOD 活性降低、细胞凋亡和 caspase-3 活性增加。此外，丹参酮恢复了 MPP 诱导的 PI3K/Akt 通路失活。此外，用 PI3K/Akt 通路抑制剂 LY294002 处理可逆转丹参酮对 SH-SY5Y 细胞和原代神经元中 MPP 诱导的 ROS 生成和凋亡的抑制作用。综上所述，丹参酮通过激活 PI3K/Akt 通路，减轻 MPP 诱导的 PD 细胞模型中 ROS 依赖性凋亡。
