Key Laboratory of Basic Pharmacology of Guizhou Province and School of Pharmacy, Zunyi Medical University, Zunyi 563000, China.
Key Laboratory of Basic Pharmacology of Ministry of Education and Joint International Research Laboratory of Ethnomedicine of Ministry of Education, Zunyi Medical University, Zunyi 563000, China.
Molecules. 2023 Apr 13;28(8):3423. doi: 10.3390/molecules28083423.
To identify potential drug candidates, secondary metabolites of were performed. As a result, two previously undescribed phenanthrene derivatives with a spirolactone ring ( and ), along with four known compounds, --cinnamoyltyramine (), ---coumaroyltyramine (), --feruloyltyramine (), and moscatilin (), were isolated from . The structures of the undescribed compounds were elucidated using NMR spectroscopy, electronic circular dichroism (ECD) calculations, and extensive spectroscopic data analysis. The cytotoxic effects of compounds on human tongue squamous cells OSC-19 were determined using MTT at concentrations of 2.5 μM, 5 μM, 10 μM, and 20 μM. Compound exhibited potent inhibitory activity against OSC-19 cells with an IC of 1.32 μM. Migration assays and western blot assays demonstrated that compound effectively inhibited migration by down-regulating MMP2 and MMP9 at concentrations of 0.5 μM and 1 μM. To investigate its effect on apoptosis, we performed AO/PI staining, flow cytometry, and WB experiments. The results showed that increasing concentrations led to increased red fluorescence, decreased green fluorescence, increased apoptosis rate, decreased expression of bcl-2, caspase 3, caspase 9, and parp proteins, and increased bax expression. Furthermore, the phosphorylation of JNK and P38 was activated, suggesting that compound may induce apoptosis via the MAPK pathway.
为了鉴定潜在的药物候选物，对 进行了次生代谢产物分析。结果，从 中分离得到了两个以前未描述的具有螺内酯环的菲衍生物（ 和 ），以及四个已知化合物——肉桂酰酪胺（）、——咖啡酰酪胺（）、——阿魏酰酪胺（）和 Moschatilin（）。使用 NMR 光谱、电子圆二色性（ECD）计算和广泛的光谱数据分析阐明了这两个未知化合物的结构。通过 MTT 在 2.5 μM、5 μM、10 μM 和 20 μM 的浓度下测定了化合物对人舌鳞癌细胞 OSC-19 的细胞毒性作用。化合物 对 OSC-19 细胞具有很强的抑制活性，IC 为 1.32 μM。迁移实验和 Western blot 分析表明，化合物 能有效抑制 MMP2 和 MMP9 的表达，从而抑制细胞迁移，在 0.5 μM 和 1 μM 的浓度下。为了研究其对细胞凋亡的影响，我们进行了 AO/PI 染色、流式细胞术和 WB 实验。结果表明，随着浓度的增加，红色荧光增加，绿色荧光减少，细胞凋亡率增加，bcl-2、caspase 3、caspase 9 和 parp 蛋白的表达减少，Bax 蛋白的表达增加。此外，JNK 和 P38 的磷酸化被激活，表明化合物 可能通过 MAPK 途径诱导细胞凋亡。
