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目的:太白银莲花皂苷B(Anemone taipaiensis saponin B)是第一次从太白银莲花中经过系统化学分析和分离鉴定的皂苷之一,所以它的生物学效应目前仍然不清楚。在本研究中,我们首次体外研究太白银莲花皂苷B对胶质瘤细胞系的生物学效应,观察它对胶质瘤细胞增殖的的抑制作用。方法:采用四甲基偶氮唑蓝(MTT)法测定太白银莲花皂苷B对胶质瘤细胞生长曲线的影响,Hoechst 33342细胞核染色后荧光显微镜观察,采用光学显微镜观察细胞的形态学变化。结果:MTT实验结果显示太白银莲花皂苷B对胶质瘤细胞U87MG和U251MG有强烈的生长抑制作用,且具有剂量依赖性,应用SPSS18.0统计软件得出太白银莲花皂苷B对U87MG细胞72 h的抑制浓度为IC25=5.2μmol/L,IC50=6.7μmol/L and IC75=8.7μmol/L,U251细胞的抑制浓度为IC25=6.2μmol/L,IC50=7.9μmol/L and IC75=10.5μmol/L。Hoechst 33342细胞核染色荧光显微镜观察以及光学显微镜下细胞形态观察显示出典型的凋亡细胞形态学特征,经过皂苷B处理后,细胞皱缩成圆球形,细胞核碎裂或者致密浓染,向核膜边缘聚集,染色质浓缩为半月状、车轮状或者马蹄状,凋亡小体出现。这些特征在24 h时更明显。结论:体外实验初步显示,太白银莲花皂苷B对U87MG和U251MG细胞具有明显的增殖抑制作用,并具有促凋亡作用。
OBJECTIVE: Anemone taipaiensis saponin B is one of the first saponins that has been systematically analyzed and isolated from the root of Chinese white lotus so its biological effect remains unclear. In this study, for the first time in vitro, we investigated the biological effects of Taibai saponin B on glioma cell lines and observed its inhibitory effect on the proliferation of glioma cells. Methods: MTT assay was used to determine the effect of saponin B on the growth of glioma cells. Hoechst 33342 cells were stained with fluorescence microscope and the morphological changes were observed by light microscope. Results: MTT assay showed that the growth of U87MG and U251MG cells was inhibited by Taibai saponin B in a dose-dependent manner. SPSS18.0 software was used to determine the effect of saponin B on U87MG cells 72 h The inhibitory concentrations of U251 cells were IC25 = 6.2μmol / L, IC50 = 7.9μmol / L and IC75 = 10.5μmol / L for IC25 = 5.2μmol / L, IC50 = 6.7μmol / L and IC75 = 8.7μmol / L. Hoechst 33342 staining by fluorescence microscopy and morphological observation of cells under light microscope showed the morphological characteristics of apoptotic cells. After saponin B treatment, the cells collapsed into a spherical, nuclear fragmentation or dense stained, to the edge of the nuclear membrane Aggregation, chromatin condensation half moon-like, wheel-shaped or horseshoe-shaped, apoptotic bodies appear. These features are more pronounced at 24 h. Conclusion: Preliminary in vitro experiments show that Taibai Yinhua saponin B has a significant inhibition of proliferation of U87MG and U251MG cells, and has a pro-apoptotic effect.