柴胡皂苷d对人肝细胞L-O2体外毒性机制探讨

Hepatotoxicity and its mechanism of saikosaponin d on the human liver L-O2 cells in vitro

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、DNAladder、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2.44μmol/L,5μmol/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。

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.