摘要
目的:建立使用微载体进行LO2肝细胞系高密度培养的方法,探讨雷公藤内酯醇的肝毒性作用及机制。方法:在限制贴壁的条件下,以Cytodex-3为材料进行LO2肝细胞的三维立体培养,在光镜下观察其生长情况,探究雷公藤内酯醇对肝细胞增殖能力的影响,并对细胞上清液的丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)、超氧化物歧化酶(SOD)和丙二醛(MDA)以及裂解细胞样品的谷胱甘肽过氧化物酶(GSH-Px)等指标进行检测。结果:雷公藤内酯醇100 g·L-1组可明显抑制肝细胞的增殖,降低肝细胞活性,使细胞ALT,AST,LDH的活性升高,MDA含量升高,SOD和GSH-Px活性降低。结论:雷公藤内酯醇对肝细胞具有明显的损伤作用且随剂量增加具有一定的剂量依赖性,作用机制与其所致细胞脂质过氧化有关。
Objective： To explore a method for high density cultivation of LO2 hepatocytes using microcarrier, and study the hepatotoxicity and its mechanism of triptolide＇s major toxic components. Method ： With the condition of the cells restricted to attach dish, three-dimensional cultivating of LO2 hepatocytes was carried out by Cytodex-3. The cell growth was observed on the light microscope and the effect of triptolide on liver cell proliferation was detected. Concentrations of alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, lactic dehydrogense （LDH）, superoxide dismutase （SOD） and malondialdehyde （MDA） in the supernatant were examined. Content of glutathione peroxidase （GSH-Px） was also examined in the cell lysate. Result： The proliferation of hepatocytes was considerably inhibited and the activity of hepatocytes was decreased by triptolide （100 μg. mL^-l）. On the other hand, the activity of ALT, AST, LDH was increased by triptolide （100 μg.mL^-1）. The content of MDA was increased, and the activities of SOD and GSH-Px were decreased by triptolide （100 μg.mL^-1）. Conclusion： The tiiptolide could do harm to hepatocytes, which may be caused by the lipid peroxidation of hepatocytes.
出处
《中国实验方剂学杂志》
CAS
北大核心
2012年第13期248-251,共4页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81072749)
教育部博士点基金项目(20103237110004)
江苏省中药药效与安全性评价重点实验室开放课题(T09004)
江苏省普通高校研究生科研创新计划
关键词
雷公藤
肝毒性
微载体黏附培养法
Tripteryginum wilfordii
hepatotoxicity
microcarrier culture
