Gao Bo, Shi Hai-Lian, Li Xiang, Qiu Shui-Ping, Wu Hui, Zhang Bei-Bei, Wu Xiao-Jun, Wang Zheng-Tao
Department of Pharmacognosy, China Pharmaceutical University, Nanjing, China.
Shanghai Key Laboratory of Complex Prescription, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai, China.
Life Sci. 2014 Jul 17;108(2):63-70. doi: 10.1016/j.lfs.2014.05.010. Epub 2014 May 23.
This study aims to investigate the effect and the mechanisms of notoginsenoside Ft1, a natural compound exclusively found in P. notoginseng, on the proliferation and apoptosis of human neuroblastoma SH-SY5Y cells.
CCK-8 assay was used to assess the cell proliferation. Flow cytometry was performed to measure the cell cycle distribution and cell apoptosis. Hoechst 33258 staining was conducted to confirm the morphological changes of apoptotic cells. Protein expression was detected by western blot analysis and caspase 3 activity was measured by colorimetric assay kit.
Among the saponins examined, Ft1 showed the best inhibitory effect on cell proliferation of SH-SY5Y cells with IC50 of 45μM. Ft1 not only arrested the cell cycle at S, G2/M stages, but also promoted cell apoptosis, which was confirmed by Hoechst 33258 staining. Further studies demonstrated that Ft1 up-regulated the protein expressions of cleaved caspase 3, phospho-p53, p21, and cyclin B1, but down-regulated that of Bcl-2. Moreover, Ft1 enhanced the phosphorylation of ERK1/2, JNK and p38 MAPK. However, the phosphorylation of Jak2 and p85 PI3K was reduced by Ft1. Inhibitors of p38 MAPK and ERK1/2 but not JNK abrogated the up-regulated protein expressions of cleaved caspase 3, p21 and down-regulated protein expression of Bcl-2 as well as elevated caspase 3 activity induced by Ft1.
Ft1 arrested the proliferation and elicited the apoptosis of SH-SY5Y cells possibly via p38 MAPK and ERK1/2 pathways, which indicates the potential therapeutic effect of it on human neuroblastoma.
本研究旨在探讨三七中特有的天然化合物三七皂苷Ft1对人神经母细胞瘤SH-SY5Y细胞增殖和凋亡的影响及其机制。
采用CCK-8法评估细胞增殖。通过流式细胞术检测细胞周期分布和细胞凋亡。进行Hoechst 33258染色以确认凋亡细胞的形态变化。通过蛋白质印迹分析检测蛋白质表达，并使用比色测定试剂盒测量caspase 3活性。
在所检测的皂苷中，Ft1对SH-SY5Y细胞的增殖具有最佳抑制作用，IC50为45μM。Ft1不仅使细胞周期停滞在S期、G2/M期，还促进细胞凋亡，这通过Hoechst 33258染色得到证实。进一步研究表明，Ft1上调了裂解的caspase 3、磷酸化p53、p21和细胞周期蛋白B1的蛋白质表达，但下调了Bcl-2的表达。此外，Ft1增强了ERK1/2、JNK和p38 MAPK的磷酸化。然而，Ft1降低了Jak2和p85 PI3K的磷酸化。p38 MAPK和ERK1/2而非JNK的抑制剂消除了Ft1诱导的裂解caspase 3、p21上调的蛋白质表达、Bcl-2下调的蛋白质表达以及caspase 3活性升高。
Ft1可能通过p38 MAPK和ERK1/2途径抑制SH-SY5Y细胞的增殖并引发其凋亡，这表明它对人神经母细胞瘤具有潜在的治疗作用。
