Hisbiscus sabdariffa Linn or Roselle is traditional herbs are widely used to treat various diseases. Roselle is rich with natural antioxidant agents such as anthocyanins, vitamin C, protocatechuic acid (PCA) and others. High oxygen reactive species (ROS) such as hidrogen peroxide (H2O2) cause oxidative stress on rat red blood cell (RBC) membrane and lead to hemolysis. The purpose of this in vitro study is to evaluate the effect of aqueous extract calyx Roselle pre-treated on rats RBC membrane towards oxidative stress induced by H2O2. Aqueous extract of calyx Roselle are used on three difference doses of concentration of 0.5, 1.0 and 2.0 mg/ml. The result showed that aqueous extract of calyx Roselle at 0.5 mg/ml concentration as a screening dose was the safest dose based on the reduction about 1.21% of hemolysis. The value was almost near to 1.19% of 0.9% NaCl group (negative control). Furthermore, at dose of 0.5 mg/ml aqueous extract of calyx Roselle studied revealed about 53% reduction via RBC percentage hemolysis when induced by 10 mM H2O2. The study was also measured the concentration of malondialdehyde (MDA), glutathione (GSH) and superoxide dismutase (SOD) activity towards 10 mM H2O2 induction. MDA concentration and SOD activity shows significantly increase as well as the decrease of GSH activity at p value <0.001 compare to 0.9% NaCl control group. Meanwhile, the study on pre-treated RBC with 0.5 mg/ml aqueous extract of calyx Roselle indicated reduction of MDA concentration and SOD activity significantly reduced (p<0.05) whereas GSH activity also significantly increase (p<0.05) compare with control group. In conclusion, this study found that concentration of dose 0.5 mg/ml aqueous extract of calyx Roselle provide a protective effect against oxidative stress induced damage on RBC membrane by 10 mM H2O2 induction.
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