Juglone In Vitro Induced By The Ls-174t Cell Apoptosis Preliminary Study

Juglone, one of important substances isolated from Juglandaceae walnut plant nuts (Jugland mandshurica Maxim)and walnut,has anti-tumour,anti-inflammatory, antibacterial and other pharmacological effects. With the continuous improvement of chemical analysis technology and the application of molecular biology technology in pharmacology, some progress has been made in the chemical and pharmacological research of Juglone. In this paper, the effect of Juglone on LS-174T cell growth and the mechanism of effects were investigated.The methods used include MTT method, fluorescence microscopical observation,Western blot and protein activity analyse.The results showed that juglone has a significantly cytotoxic activity on LS-174T cell.The studies demonstrate that juglone inhibited the proliferation of LS-174T cells in a time and dose-dependent manner. Fluorochrome stain assay certified that juglone can induce apoptosis of LS-174T cells.After 24h exposured to juglone (25μM and 50μM), the cells presented the typical morphological changes of apoptosis.The activity of Caspase rised to the highest at 24h then descended.Caspase-3 activity showed a dose dependent but Caspase-9 didn't. After cells were treated with juglone for 36h, the expression of Bcl-2 was down-regulated and the expression of Caspase active fragment was up-regulated compared to the control.The expression of p53 and p21 have no change.GSH and NAC can inhibit the apoptosis,p<0.001 compared with the control group. At the same time the expression of Bcl-2 was up-regulated and the expression of Caspase active fragment was down-regulated.In conclusionjuglone may induce apoptosis in LS-174 cells through a mitochondrial pathway.Caspase and the generation of ROS may participate the pathway.