1) Objective
This study was done in order to investigate the antioxidative effects of SR(Scutellariae Radix) aqua-acupuncture extract solution on lipid peroxidation by free radicals
2) Material and Methods
At inervals during autoxidation of linoleic acid in the water-alcohol system, the degree of oxidation was measured by the TBA method. The reaction mixture contained 50㎕ of sample, 0.2㎖ of 8.1% sodium dodecylsulfate(SDS), 1.5㎖ of 20% acetic acid solution(pH 3.5), and 1.5㎖ of 0.8% aqueous solution of TBA. The pH of 20% acetic acid solution was adjsted with 10N NaOH. The mixture was finally made up to 4.0㎖ with distilled water, and placed at 5℃ for 60min, and then heated at 95℃ for 60min. After cooling with tap water, the absorbance was measured at 532㎚.
The effects of SR aqua-acupuncture extract solution on DPPH radical was determined according to the method of Hatano, SR aqua-acupuncture extract solution in 4㎖ of distilled water were added to a methanolic solution of DPPH(1mM, 1㎖). The mixture was shaken and left to stand at room temperature for 30min ; the absorbance of the resulting solution was measured spectrophotometrically at 517㎚. Each values are the mean of triplicate experiments. In addition, to evaluate the safety of SR aqua-acupuncture extract solution on normal rat liver cell(Ac2F), the cells(5×10⁴cells/well) were plated on 75-㎠ plastic flasks in 20㎖ of DMEM, 10% heat-inactivated fetal bovine serum. And cells were incubated under 5% CO₂ 95% air, 37℃ for 2hrs. After washing, various concentration of SR aqua-acupuncture extract solution were added, and incubated for 18∼20hus. Viable cells were detected by MTT assay. All data are the mean of triplicated determination And, to investigate the antioxidative activity of SR aqua-acupuncture extract solution on tert-butyl hydroperoxide (t-BHP) induced cultured normal rat liver cell death, cells(5×10⁴cells/well) were incubated under 5% CO₂, 95% air, at 37℃ for 2hrs. After washing, various concentration of SR aqua-acupuncture extract solution were added, and incubated for 18∼20hrs. After preincubation, t-BHP(final concentration 1mM) was added, and the reaction mixture was incubated for 2hrs. And then, viable cells were detected by MTT assay. All data are the mean of triplicated determination.
3) Conclusion
SR aqua-acupuncture extract solution exhibited markedly antioxidant activity, which inhibited linoleic acid peroxidation. And SR aqua-acupuncture extract solution showed 70.23% scavenging effect on DPPH radical. Inhibitery effect of SR aqua-acupuncture extract solution on the peroxidation of rat liver homogenate by hydroxyl radical(OH), derived from hydrogen peroxidase-Fe^(2+) system. SR aqua-acupuncture extract solution protected the cell death induced by t-BHP and significantly increased cell viability in normal rat liver cell(Ac2F). These results suggested that SR aqua-acupuncture extract solution might play a protective role in lipid peroxidation by free radical.
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