title: Activation of apoptosis by ethyl acetate fraction of ethanol extract of Dianthus superbus in HepG2 cell line.
abstract: Dianthus superbus L. is commonly used as a traditional Chinese medicine. We recently showed that ethyl acetate fraction (EE-DS) from ethanol extract of D. superbus exhibited the strongest antioxidant and cytotoxic activities. In this study, we examined apoptosis of HepG2 cells induced by EE-DS, and the mechanism underlying apoptosis was also investigated. Treatment of HepG2 cells with EE-DS (20-80 μg/ml) for 48 h led to a significant dose-dependent increase in the percentage of cells in sub-G1 phase by analysis of the content of DNA in cells, and a large number of apoptotic bodies containing nuclear fragments were observed in cells treated with 80 μg/ml of EE-DS for 24 h by using Hoechst 33258 staining. These data show that EE-DS can induce apoptosis of HepG2 cells. Immunoblot analysis showed that EE-DS significantly suppressed the expressions of Bcl-2 and NF-κB. Treatment of cells with EE-DS (80 μg/ml) for 48 h resulted in significant increase of cytochrome c in the cytosol, which indicated cytochrome c release from mitochondria. Activation of caspase-9 and -3 were also determined when the cells treated with EE-DS. The results suggest that apoptosis of HepG2 cells induced by EE-DS could be through the mitochondrial intrinsic pathway. High performance liquid chromatography (HPLC) data showed that the composition of EE-DS is complicated. Further studies are needed to find the effective constituents of EE-DS.
authors.affiliation: Key Laboratory of Combinatorial Biosynthesis and Drug Discovery (Wuhan University), Ministry of Education, and Institute of TCM & Natural Products, Wuhan University School of Pharmaceutical Sciences, Wuhan 430071, PR China. jqyu@whu.edu.cn
authors.forename: Jian-Qing
authors.initials: JQ
authors.lastname: Yu
authors.forename: Yan
authors.initials: Y
authors.lastname: Yin
authors.forename: Jia-Chuan
authors.initials: JC
authors.lastname: Lei
authors.forename: Xiu-Qiao
authors.initials: XQ
authors.lastname: Zhang
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Cheng-Li
authors.initials: CL
authors.lastname: Ding
authors.forename: Shan
authors.initials: S
authors.lastname: Wu
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: He
authors.forename: Yan-Wen
authors.initials: YW
authors.lastname: Liu
authors.forename: Guo-Lin
authors.initials: GL
authors.lastname: Zou
mesh_terms: D000085:Acetates / Q000737:chemistry; D017209:Apoptosis / Q000187:drug effects*; D045744:Cell Line, Tumor; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D029749:Dianthus / Q000737:chemistry*; D000431:Ethanol / Q000737:chemistry; D056945:Hep G2 Cells; D006801:Humans; D010936:Plant Extracts / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: D000085:Acetates; D010936:Plant Extracts; D000431:Ethanol; C007650:ethyl acetate