柴胡皂苷d 对人肝细胞L-O2 体外毒性机制探讨

摘要/Abstract

摘要： 目的考察柴胡主要成分之一柴胡皂苷d(ssd)体外肝毒性以及主要机制。方法通过MTT 试验、细胞形态学改变、DNA ladder 、DAPI 荧光染色法、乳酸脱氢酶释放率、溶血试验等方法考察ssd 对人肝细胞L-O2 是否具有毒性作用, 进而阐明ssd 肝毒性产生的可能机制。结果 MTT 法测得ssd 的半抑制浓度(IC50)为2.44 μmol\L, 5 μmol/L 处理组细胞形态学发生明显改变, 乳酸脱氢酶释放率增高和溶血试验结果发现毒性作用呈剂量依赖性, 但DNAladder 和DAPI 荧光检测均未见明显细胞凋亡。结论 ssd 具有较强的体外肝毒性, 其机制可能是由于ssd 诱导细胞膜通透性增加从而导致细胞损伤或坏死, 而不是诱导细胞凋亡。

关键词: 柴胡皂苷d, L-O2 肝细胞, 体外肝毒性

Abstract: AIM: This study was to determine whether saikosaponin d (ssd)caused hepatotoxicity and to investigate the possible toxicity mechanism. METHODS: Experiment was carried out using the human live cell lines (L-O2)in vitro.MTT assay, change of cellular morphology, DNA fragmentation analysis, nuclear staining with 4, 6-diamidino-2-phenylindole (DAPI), hemolysis assay and membrane leakage of lactate dehydrogenase (LDH assay) were adopted to evaluate the hepatotoxicity of ssd. RESULTS: Results showed that L-O2 cells were inhibited significantly by above 2.5 μmol/L ssd, and IC50 = 2.44 μmol L.The change of celluar morphology, increase of LDH release and hemolytic ratio (%)could be detected significantly when treated with 5 μmol/L ssd. However, other results showed no obvious induction on the apoptosis from DNA ladder and DAPI. CONCLUSION: The toxicity mechanism of ssd on the L-O2 cells in vitro might be not through apoptosis, but related to the cytolysis.

Key words: saikosaponin d, huamn liver cell L-O2, hepatotoxicity in vitro

中图分类号:

R965.2
R966

李涛, 江振洲, 王涛, 张陆勇, 徐晓月, 贾晓明, 缪文英, 闵超. 柴胡皂苷d 对人肝细胞L-O2 体外毒性机制探讨[J]. 中国临床药理学与治疗学, 2007, 12(4): 396-400.

LI Tao, JIANG Zhen-zhou, WANG Tao, ZHANg/Lu-yong, XU Xiao-yue, JIA Xiao-ming, MIAO Wen-ying, MIN Chao. Hepatotoxicity and its mechanism of saikosaponin d on the human liver LO2cells in vitro[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2007, 12(4): 396-400.

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