摘要
目的观察黄芩配方颗粒(SBFG)对人肝癌Hep G2细胞增殖及凋亡的影响,并探讨其作用机制。方法细胞分为正常对照组,0.05、0.1、0.2、0.4、0.8、1.6 g/L SBFG组,共7组,不同浓度SBFG作用于Hep G2细胞24 h,采用四甲基偶氮唑蓝(MTT)法,观察SBFG对Hep G2细胞增殖的影响;Hoechst 33342染色法观察细胞凋亡情况;高内涵分析(HCS)技术检测线粒体膜电位(MMP)、细胞色素C、膜通透性、核面积、核DNA含量、细胞数量的变化。结果 SBFG可明显抑制Hep G2细胞增殖(P<0.01),抑制率呈浓度依赖性,IC50值为(0.38±0.016)g/L;Hoechst 33342染色可见细胞核浓染、核质浓缩、核破碎等细胞凋亡的形态学变化;高内涵分析也发现,0.1 g/L SBFG组细胞MMP丢失以及细胞色素C释放量明显增加(P<0.01),0.4 g/L SBFG组进一步显著降低MMP、增加细胞色素C含量,并可使细胞膜通透性显著升高(P<0.01)、核固缩及核荧光强度显著增强(P<0.01),1.6 g/L SBFG组孵育细胞,除上述凋亡特征外,还伴随着细胞数量的直接下降(P<0.01)。结论 SBFG可抑制人肝癌Hep G2细胞增殖,促进Hep G2细胞凋亡,其机制可能与降低MMP、促进细胞色素C释放、增加细胞膜通透性等有关。
Objective To investigate the effect and machanism of Scutellaria baicalensis formula granule（ SBFG） on proliferation and apoptosis of Hep G2 human hepatoma cells. Methods Hep G2 cells were divided into normal control group and six SBFG groups at concentration of 0. 05,0. 1,0. 2,0. 4,0. 8 and1. 6 g / L. After Hep G2 cells were treated by SBFG for 24 hours,the proliferation was analyzed by using MTT method,apoptosis was observed by Hoechst 33342 staining method,and the changes of cell count,nuclear size,nuclear DNA content,permeability,mitochondria membrane potential（ MMP） and cytochrome C were tested by high content screening assays（ HCS）. Results The inhibitory effect of SBFG on proliferation of Hep G2 cells was significant（ P〈0. 01） with concentration-dependence,and IC50of（ 0. 38 ± 0. 016） g /L. By Hoechst 33342 staining,the morphological changes were observed including hyperchromatic nuclei,concentrated karyoplasms and karyorrhexis. Furthermore,HCS indicated that 0. 1g / L SBFG decreased MMP and increased cytochrome C release significantly（ P〈0. 01）; 0. 4 g / L SBFG further decreased MMP and increased cytochrome C release,and increased the membrane permeability significantly,pyknosis and nuclear fluorescence intensity（ P〈0. 01）; in addition to the effects above,1. 6 g / L SBFG also reduced the number of cells（ P〈0. 01）. Conclusion SBFG can significantly inhibit the proliferation of Hep G2 cells,and induce cell apoptosis. The mechanisms may be related to decreasing the MMP,promoting the release of cytochrome C and increasing membrane permeability.
作者
任艳青
田宇柔
牛丽颖
王鑫国
REN Yanqing TIAN Yurou NIU Liying WANG Xinguo(Pharmacological Lab of Traditional Chinese Medicine, Hebei University of Chinese Medicine, Hebei TCM Formula Granule Engineering ＆Technology Research Center, Hebei 050091)
出处
《北京中医药大学学报》
CAS
CSCD
北大核心
2016年第10期844-849,共6页
Journal of Beijing University of Traditional Chinese Medicine
基金
河北省高等学校科学技术研究项目(No.ZD2015001)
关键词
黄芩配方颗粒
人肝癌HEPG2细胞
细胞增殖
细胞凋亡
高内涵分析
Scutellaria baicalensis formula granule
human hepatoma cancer cell line HepG2
cell proli feration
apoptosis
high content screening assay
