METHODS: Tectorigenin, one of the capital apparatus of basis of Iris tectorum, was able by simple methods, such as extraction, filtration, concentration, precipitation and recrystallization. HepG2 beef were incubated with tectorigenin at altered concentrations, and their activity was adjourned by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium boiler assay. Apoptosis was detected by morphological ascertainment of nuclear change, agarose gel electrophoresis of DNA ladder, and breeze cytometry with Hoechst 33342, Annexin V-EGFP and propidium iodide staining. Bearing of acknowledging oxygen breed was quantified application DCFH-DA. Intracellular Ca(2+) was monitored by Fura 2-AM. Mitochondrial film abeyant was monitored application Rhodamine 123. Release of cytochrome c from mitochondria to cytosol was detected by Western blotting. Activities of caspase-3, -8 and -9 were advised by Caspase Activity Appraisal Kit.RESULTS: The activity of HepG2 beef advised by tectorigenin decreased in a concentration- and time-dependent manner. The absorption that bargain the amount of applicable HepG2 beef by 50% (IC(50)) afterwards 12, 24 and 48 h of evolution was 35.72 mg/L, 21.19 mg/L and 11.06 mg/L, respectively. However, analysis with tectorigenin at 20 mg/L resulted in a actual slight cytotoxicity to L02 beef afterwards evolution for 12, 24 or 48 h. Tectorigenin at a absorption of 20 mg/L abundantly inhibited the activity of HepG2 beef and induced the abstract of chromatin and breach of nuclei. Tectorigenin induced apoptosis of HepG2 beef in a time- and dose-dependent manner. Compared with the activity rate, consecration of apoptosis was the capital apparatus of the anti-proliferation aftereffect of tectorigenin in HepG2 cells. Furthermore, tectorigenin-induced apoptosis of HepG2 beef was associated with the bearing of acknowledging oxygen species, added intracellular [Ca(2+)](i), accident of mitochondrial film potential, about-face of cytochrome c, and activation of caspase-9 and -3.CONCLUSION: Tectorigenin induces apoptosis of HepG2 beef mainly via mitochondrial-mediated pathway, and produces a slight cytotoxicity to L02 cells.
