Department of Pharmacy, Medical College, Jianghan University, Wuhan, 430056, China.
Hubei Key Laboratory of Resources and Chemistry of Chinese Medicine, Hubei University of Chinese Medicine, Wuhan, 430065, China.
Curr Med Sci. 2020 Feb;40(1):138-144. doi: 10.1007/s11596-020-2157-5. Epub 2020 Mar 13.
This study examined anti-cancer compounds present in the chloroform extract of the Chinese medicine formula Shenqi San (CE-SS). Silica gel column chromatography, Sephadex LH-20, octadecylsilyl (ODS) column chromatography, and high performance liquid chromatography (HPLC) were used to separate the compounds from CE-SS. The structural formulas of the separated compounds were determined using 1D H and C experiments as well as high resolution electrospray ionization mass spectroscopy (HRESIMS). The corresponding results were compared with the reported literature data. A total of six compounds were separated and their structures were identified on the basis of corresponding spectroscopic and physico-chemical properties. They were Saikogenin F (I), Prosaikogenin D (II), Prosaikogenin F (III), β-sitosterol (IV), 3β,16β,23-trihydroxy-13,28-epoxyurs-11-ene-3-O-β-D-glucopyranoside (V), and methyl ursolic acid (VI). The separated compounds were evaluated in vitro for their inhibitory ability against the proliferation of A549 cells via MTT assay. Apoptosis was investigated using Annexin V-FITC/propidium iodide (PI) by flow cytometry. Apoptosis-associated proteins were examined by Western blotting. All the compounds were observed to have inhibitory activities against the proliferation of A549 cells to different degrees. Flow cytometry showed that compound V increased the proportion of apoptotic A549 cells in a dose-dependent manner. Western blotting showed that compound V increased the expression of Bax, cleaved-caspase-3, cleaved-caspase-9 and cleaved-poly ADP-ribose polymerase (PARP), and decreased the expression of Bcl-2. These results indicated that compound V featured a significant inhibitory effect on A549 cells when compared with other compounds, and it may be considered a potential drug against cancers.
本研究考察了中药参芪三（CE-SS）氯仿提取物中的抗癌化合物。采用硅胶柱层析、葡聚糖 LH-20 凝胶层析、十八烷基硅烷（ODS）柱层析和高效液相色谱（HPLC）对 CE-SS 中的化合物进行分离。通过 1D H 和 C 实验以及高分辨电喷雾电离质谱（HRESIMS）确定分离化合物的结构公式。相应的结果与文献报道的数据进行了比较。总共分离出六种化合物，并根据相应的光谱和物理化学性质确定了它们的结构。它们分别为沙克苷元 F（I）、原沙克苷元 D（II）、原沙克苷元 F（III）、β-谷甾醇（IV）、3β,16β,23-三羟基-13,28-环氧乌苏-11-烯-3-O-β-D-吡喃葡萄糖苷（V）和甲基乌苏酸（VI）。通过 MTT 法测定分离出的化合物对 A549 细胞增殖的抑制能力，评估其体外活性。通过流式细胞术用 Annexin V-FITC/碘化丙啶（PI）检测细胞凋亡。通过 Western blot 检测凋亡相关蛋白。所有化合物均观察到对 A549 细胞增殖有不同程度的抑制活性。流式细胞术结果表明，化合物 V 呈剂量依赖性地增加凋亡 A549 细胞的比例。Western blot 结果表明，化合物 V 增加了 Bax、cleaved-caspase-3、cleaved-caspase-9 和 cleaved-poly ADP-ribose polymerase（PARP）的表达，降低了 Bcl-2 的表达。这些结果表明，与其他化合物相比，化合物 V 对 A549 细胞具有显著的抑制作用，可能是一种有潜力的抗癌药物。
