"VINČA" Institute of Nuclear Sciences, National Institute of Thе Republic of Serbia, University of Belgrade, Mike Petrovića Alasa 12-14, 11351 Belgrade, Serbia.
"VINČA" Institute of Nuclear Sciences, National Institute of Thе Republic of Serbia, University of Belgrade, Mike Petrovića Alasa 12-14, 11351 Belgrade, Serbia.
Chem Biol Interact. 2024 Aug 1;398:111103. doi: 10.1016/j.cbi.2024.111103. Epub 2024 Jun 7.
Gentiopicroside (Gp) and swertiamarin (Sm), secoiridoid glycosides commonly found in plants of the Gentianaceae family, differ in one functional group. They exhibit promising cytotoxic effects in cancer cell lines and overall protective outcomes, marking them as promising molecules for developing novel pharmaceuticals. To investigate potential variations in cellular sensitivity to compounds of similar molecular structures, we analyzed the mode of Gp and Sm induced cell death in human peripheral blood mononuclear cells (PBMCs) after 48 h of treatment. The lowest tested concentration that significantly reduces cell viability, 50 μM, was applied. Oxidative stress parameters were estimated by measuring the levels of prooxidative/antioxidative balance, lipid peroxidation products, and 8-oxo-7,8-dihydro-2-deoxyguanosine, while gene expression of DNA repair enzymes was evaluated by employing quantitative real-time PCR. Cellular morphology was analyzed by fluorescent microscopy, and immunoblot analysis of apoptosis and necroptosis-related proteins was used to assess the type of cell death induced by the treatments. The discriminatory impact of Gp/Sm treatments on apoptosis and necroptosis-induced cell death was evaluated by monitoring the cell survival in co-treatment with specific cell death inhibitors. Obtained results show greater cytotoxicity of Gp than Sm suggesting that variations in the molecular structures of the tested compounds can substantially affect their biological effects. Gp/Sm co-treatment with apoptosis and necroptosis inhibitors revealed a distinct, albeit non-specific mechanism of PBMCs cell death. Although the therapeutic may not directly cause a specific type of cell death, its extent can be pivotal in assessing the safety of therapeutic application and developing phytopharmaceuticals with improved features. Since phytopharmaceuticals affect all exposed cells, identification of cytotoxic mechanisms on PBMCs after Gp and Sm treatment is important for addressing the formulation and dosage of potential phytopharmaceuticals.
龙胆苦苷（Gp）和獐牙菜苦苷（Sm）是存在于龙胆科植物中的两种环烯醚萜苷类化合物，它们仅在一个功能基团上有所不同。这两种化合物在癌细胞系中表现出有希望的细胞毒性作用和整体保护作用，这使它们成为开发新型药物的有前途的分子。为了研究具有相似分子结构的化合物对细胞敏感性的潜在差异，我们分析了在 48 小时的治疗后，Gp 和 Sm 诱导人外周血单个核细胞（PBMCs）细胞死亡的模式。应用了最低测试浓度（50 μM），该浓度显著降低细胞活力。通过测量促氧化剂/抗氧化剂平衡、脂质过氧化产物和 8-氧代-7,8-二氢-2-脱氧鸟苷的水平来估计氧化应激参数，同时通过定量实时 PCR 评估 DNA 修复酶的基因表达。通过荧光显微镜分析细胞形态，并用免疫印迹分析评估凋亡和坏死相关蛋白，以评估处理诱导的细胞死亡类型。通过监测与特定细胞死亡抑制剂共同处理时的细胞存活率，评估 Gp/Sm 处理对凋亡和坏死诱导的细胞死亡的区分作用。研究结果表明，Gp 的细胞毒性大于 Sm，这表明测试化合物的分子结构差异可以显著影响其生物学效应。Gp/Sm 与凋亡和坏死抑制剂共同处理显示 PBMCs 细胞死亡的一种独特但非特异性机制。尽管治疗剂可能不会直接导致特定类型的细胞死亡，但在评估治疗应用的安全性和开发具有改进特征的植物药方面，其程度可能至关重要。由于植物药会影响所有暴露的细胞，因此鉴定 Gp 和 Sm 处理后 PBMCs 的细胞毒性机制对于确定潜在植物药的配方和剂量非常重要。
