摘要
目的:探讨狭基线纹香茶菜水溶性总黄酮(WSTF)对氧化应激损伤LO_2细胞的保护作用。方法:通过细胞毒性实验确定给药范围,建立过氧化氢(H2O_2)致LO_2细胞损伤模型,用含不同浓度WSTF的培养液与急性损伤肝细胞共孵育不同时间,采用MTT比色法测定细胞活力,检测谷氨酸氨基转移酶(ALT)、天冬氨酸氨基转移酶(AST)和丙二醛(MDA)的水平,评价WSTF对肝细胞损伤的保护作用。检测细胞线粒体膜电位和细胞内活性氧簇(ROS)水平,分析WSTF抑制H2O_2诱导LO_2细胞凋亡情况。结果:0.3 mmol·L^(-1)H2O_2处理LO_2细胞4 h构建H2O_2损伤LO_2细胞模型,确定0.031 2~0.125 mg·ml^(-1)WSTF为保护LO_2细胞损伤的给药浓度范围。WSTF可抑制H2O_2对肝细胞的损伤,并明显降低H2O_2致急性损伤肝细胞的ALT、AST释放量和MDA水平,逆转H2O_2诱导LO_2肝细胞线粒体膜电位去极化和细胞内ROS升高。结论:WSTF体外给药对肝细胞损伤有保护作用。
Objective： To explore the protective effect of the water-soluble total flavonoids from Isodon lophanthoides var. gerardia- nus （Benth.） H. Hara on LO2 cells damage. Methods： The cytotoxicity was evaluated by MTI＂ cell viability determination to confirm the concentration range. Hepatocyte damage model was established by H2O2 treatment. After the oxidative stress hepatocyte was coin- cubated with WSTF at different concentrations for various times, the protective effect of WSTF on H2O2 -induced hepatocyte damage was evaluated by MTT cell viability determination and the content determination of ALT, AST and MDA in cell supernatant. The inhibition of WSTF against H2O2 -induced LO2 ceils apoptosis was evaluated by the quantitative determination of Rhodamine 123 fluorescence and intracellular ROS. Results： The LO2 cells injured by 0.3 mmol L-l H2O2 treatment for 4 h were used as the hepatocyte damage model. The concentration range of WSTF was 0.031 2-0. 125 mg m1-1. WSTF could inhibit H2O2- induced injury in LO2 cells and obviously reduce ALT, AST and MDA. Moreover, WSTF could reverse mitochondrial membrane potential depolarization and decrease the amount of intracellular ROS. Conclusion： WSTF exhibits notable protective and curative effects on hepatocyte damage in vitro.
出处
《中国药师》
CAS
2018年第1期53-57,共5页
China Pharmacist
基金
2016年湖南省中医药科研计划重点项目(编号:201629)
