Effect of Hydroxysafflor Yellow A on the Oxidation of Intracellular Glutathione Induced by Hydrogen Peroxide in L02 Cells

The effect of hydroxysafflor A(HSYA) on the oxidation of intracellular reduced glutathione(GSH) induced by hydrogen peroxide(H 2 O 2) was investigated in L02 cells. 5,5 '-Dithiobis(2-nitrobenzoic acid)(DTNB) was used as the derivatizing agent and the reduced glutathione was measured by HPLC. The total glutathione was measured by the same method after reduced by dithiothreitol(DTT). Oxidative glutathione can be calculated through reduced glutathione and total glutathione. The L02 cells were pretreated with HSYA for 24 hours, and then exposed to H 2 O 2 for 1 hour. The intracellular GSH, GSSG contents and GSSG/GSH were analyzed by HPLC. The results showed that the standard curve was linear over the range of 0.1-2 mmol/L. The minimum quantification limit was 0.1 mmol/L. The precision discrepancy of inter-batch and intra-batch assays were both less than 10%. The recovery rates at low, medium and high spiked concentrations were in the range of 95%-105%. The developed method proved to accurate, quick, precise and suitable for the measurement of glutathione in cells. HSYA can significantly inhibit the increase of GSSG and GSSG/GSH, thus relieving the oxidative stress induced by H 2 O 2.