Abstract: Objective: To observe the effects of blocking and activating chloride channels on hemolysis induced by puerarin injection in rabbits and to investigate the roles of chloride channels in hemolytic reaction induced by puerarin injection. Methods: Rabbit erythrocyte suspension was incubated with different concentrations of puerarin injection(0.75, 1.5, 3, 6, 12 mg/ml) at 37^oC for 6 hours. The cell imaging system was employed to observe whether puerarin injection induced hemolysis. The hemolysis rate was detected by microplate reader and flow cytometry. Effects of activating and closing chloride channels on the hemolysis induced by puerarin injection were explored. Results: Puerarin injection could induce the hemolysis of rabbit erythrocytes in vitro. In the range of 1.5 mg/ml~12 mg/ml, puerarin injection could induce hemolysis in a concentration-dependent manner (n=3, P<0.01). The chloride channel blockers tamoxifen (20 μmol/L) and ATP (10 mmol/L) significantly inhibited the hemolysis induced by puerarin injection (n=3~5, P<0.01). Application of low concentration ATP (50 μmol/L) to activate the chloride channel significantly increased puerarin injection induced hemolysis (n=4, P<0.01). Conclusion: The hemolytic effect of puerarin injection is dose-dependent in vitro, and the activation of chloride channel is closely related to the hemolysis induced by puerarin injection.

Key words: chloride channel, puerarin injection, erythrocyte, hemolysis, rabbit