Jiang Zequn, Ma Yanxia, Zhou Lihua, Jiang Haiying, Wang Mingyan, Zhan Xiuqin
Department of Preclinical Medicine, Nanjing University of Chinese Medicine, Nanjing, People's Republic China.
J Intercult Ethnopharmacol. 2014 Oct-Dec;3(4):201-5. doi: 10.5455/jice.20140916011549. Epub 2014 Sep 27.
The aim was to determine the action mode of cornel iridoid glycoside (CIG) from Fructus corni on hepatoprotective activities, the effects of CIG on human hepatocyte cell line (L02) injured by D-galactosamine (GalN) and tumor necrosis factor-α (TNF-α) were examined.
The percentage of cell viability was evaluated by cell counting kit-8 assay. Apoptosis was detected by flow cytometric analysis in human L02 hepatocytes. The expression levels of activating transcription factor-4 (ATF4), and C/EBP homologous protein (CHOP) were detected by western-blot analysis. In addition, the activity of caspase-3 was tested by enzyme-linked immunosorbent assay.
The results showed that CIG caused a significant increase in the viability of L02 cells injured by GalN/TNF-α, in accordance with a dose-dependent decrease of apoptotic cell death confirmed by flow cytometric analysis. Based on western blot and colorimetric assay, we found that GalN/TNF-α induced increased expression of ATF4, CHOP, and activation of caspase-3 while CIG pre-treatment had a dose-dependent suppression on them in this cell model.
Overall, these findings demonstrate that CIG can effectively protect L02 hepatocytes against apoptosis induced by GalN/TNF-α, suggesting that it is a possible candidate target for liver disease therapy.
本研究旨在确定山茱萸中的山茱萸环烯醚萜苷（CIG）的保肝作用模式，检测CIG对D-氨基半乳糖（GalN）和肿瘤坏死因子-α（TNF-α）损伤的人肝细胞系（L02）的影响。
采用细胞计数试剂盒-8法评估细胞活力百分比。通过流式细胞术分析检测人L02肝细胞中的细胞凋亡情况。采用蛋白质免疫印迹分析检测激活转录因子4（ATF4）和C/EBP同源蛋白（CHOP）的表达水平。此外，通过酶联免疫吸附测定法检测caspase-3的活性。
结果显示，CIG可显著提高GalN/TNF-α损伤的L02细胞的活力，流式细胞术分析证实其可剂量依赖性地减少凋亡细胞死亡。基于蛋白质免疫印迹和比色测定，我们发现GalN/TNF-α可诱导ATF4、CHOP表达增加以及caspase-3激活，而在该细胞模型中，CIG预处理对它们具有剂量依赖性抑制作用。
总体而言，这些研究结果表明，CIG可有效保护L02肝细胞免受GalN/TNF-α诱导的细胞凋亡，提示其可能是肝病治疗的候选靶点。
