de Beer A D, Rudolph W, Maharaj V, Steenkamp V, Balmith M, Cordier W
Department of Pharmacology, Faculty of Health Sciences, University of Pretoria, South Africa.
Biodiscovery Centre, Department of Chemistry, Faculty of Natural and Agricultural Sciences, University of Pretoria, South Africa.
J Ethnopharmacol. 2026 May 10;362:121222. doi: 10.1016/j.jep.2026.121222. Epub 2026 Feb 7.
Ferroptosis contributes to Parkinson's disease progression given dysregulation of iron homeostasis and redox status. Polygala virgata is used ethnomedicinally for memory enhancement. This study assessed the cytoprotective and antioxidant properties of crude extracts and fractions of P. virgata using a 6-hydroxydopamine-induced (6-OHDA) SH-SY5Y neuroblastoma cytotoxicity model.
Dried roots of P. virgata (14% w/v) were sequentially extracted using dichloromethane/methanol (1:1) and methanol, which was combined to give a crude extract. The crude extract was separated into seven fractions using different ratios of water, acetonitrile and methanol on solid phase extraction (SPE). Inherent cytotoxicity of the samples (10 μg/mL), as well as their ability to reduce 6-OHDA-induced cytotoxicity (35 μM), was determined using the sulforhodamine B (SRB) assay after 48-h (h) exposure. The active fractions' cytoprotective effect in relation to reactive oxygen species (ROS), glutathione levels (GSH), lipid peroxidation, and mitochondrial integrity was determined fluorometrically. Cytoprotective fractions' phytochemical constituency was elucidated using liquid chromatography high resolution mass-spectrometry (UPLC-HRMS).
Fractions 3 to 7 increased cell density after exposure to 6-OHDA by 31.14%, 28.08%, 30.72%, 40.58% (p < 0.01) and 28.86%, respectively, with no inherent cytotoxicity observed. Fraction 4 reduced 6-OHDA-induced ROS generation (2.09-fold) and lipid peroxidation (0.28-fold). Non-significant increases in GSH were noted (1.34 to 19.25%), while all fractions hyperpolarised the mitochondrial membrane. Multi-hydroxylated xanthones, flavones and flavans were tentatively identified using UPLC-HRMS.
P. virgata fractions reduced 6-OHDA-induced cytotoxicity via decreased oxidative stress and hyperpolarisation of the mitochondrial membrane, most likely ascribed to the identified xanthones, flavones and flavans. Isolation and purification of these compounds are warranted as potential antioxidant scaffolds.
鉴于铁稳态和氧化还原状态的失调，铁死亡促进帕金森病的进展。黄花远志在民族医学中用于增强记忆力。本研究使用6-羟基多巴胺诱导（6-OHDA）的SH-SY5Y神经母细胞瘤细胞毒性模型，评估了黄花远志粗提物及其各部位的细胞保护和抗氧化特性。
将黄花远志干燥根（14% w/v）依次用二氯甲烷/甲醇（1:1）和甲醇提取，合并后得到粗提物。粗提物在固相萃取（SPE）上使用不同比例的水、乙腈和甲醇分离成七个部位。在暴露48小时后，使用磺酰罗丹明B（SRB）测定法测定样品（10μg/mL）的固有细胞毒性及其降低6-OHDA诱导的细胞毒性（35μM）的能力。通过荧光法测定活性部位对活性氧（ROS）、谷胱甘肽水平（GSH）、脂质过氧化和线粒体完整性的细胞保护作用。使用液相色谱高分辨率质谱（UPLC-HRMS）阐明细胞保护部位的植物化学成分。
部位3至7在暴露于6-OHDA后细胞密度分别增加了31.14%、28.08%、30.72%、40.58%（p<0.01）和28.86%，未观察到固有细胞毒性。部位4减少了6-OHDA诱导的ROS生成（2.09倍）和脂质过氧化（0.28倍）。观察到GSH有不显著增加（1.34%至19.25%），而所有部位均使线粒体膜超极化。使用UPLC-HRMS初步鉴定出多羟基呫吨酮、黄酮和黄烷。
黄花远志部位通过降低氧化应激和线粒体膜超极化减少6-OHDA诱导的细胞毒性，这很可能归因于所鉴定的呫吨酮、黄酮和黄烷。作为潜在的抗氧化支架，对这些化合物进行分离和纯化是有必要的。
