Effect of quercetin 7-rhamnoside on glycochenodeoxycholic acid-induced L-02 human normal liver cell apoptosis

摘要

Quercetin 7-rhamnoside (Q7R) is one of the main flavonoid components of Hypericum japonicum. However, whether Q7R is one of the active ingredients responsible for the hepatopreventive effects of Hypericum japonicum has not yet been ascertained. Thus, the aim of the present study was to elucidate whether Q7R attenuates apoptosis induced by glycochenodeoxycholic acid (GCDC) in vitro, and to elucidate the mechanisms involved. L-02 human normal liver cells were pre-incubated with 0, 50, 100 and 200 mu M Q7R for 30 min and then exposed to 100 mu M GCDC for the indicated periods of time. Methylthiazolyldiphenyl-tetrazolium bromide (MTT) was performed to examine cell viability. Apoptosis was evaluated by Hoechst 33258 staining and Annexin V-FITC/PI double staining. Intracellular reactive oxygen species (ROS) were detected by flow cytometry using the oxidation-sensitive fluorescent probe, DCFH-DA. The assay for glutathione (GSH) was performed using a GSH detection kit. Intracellular Ca2+ concentration was evaluated using a confocal laser scanning microscope with Fluo-3 as the Ca2+ probe and mitochondrial membrane potential (Delta psi m) was measured by rhodamine 123 (Rh123) fluorescence. Q7R attenuated the GCDC-induced reduction in cell viability and the high apoptotic rate. Moreover, Q7R protected the L-02 cells from ROS overproduction, GSH depletion, intracellular Ca2+ accumulation and Delta psi m decrease induced by GCDC. These results suggest that Q7R attenuates L-02 cell injury induced by GCDC, possibly by inhibiting the overproduction of ROS, GSH depletion, intracellular Ca2+ accumulation and Delta psi m decrease, thereby minimizing L-02 cell apoptosis.

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