[Effects of chloride channels on hemolysis induced by puerarin injection].

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 37C 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 the range of 1.5 mg/ml12 mg/ml, puerarin injection could induce hemolysis in a concentration-dependent manner (=3, <0.01). The chloride channel blockers tamoxifen (20 μmol/L) and ATP (10 mmol/L) significantly inhibited the hemolysis induced by puerarin injection (=35, <0.01). Application of low concentration ATP (50 μmol/L) to activate the chloride channel significantly increased puerarin injection induced hemolysis (=4, <0.01).

CONCLUSIONS

The hemolytic effect of puerarin injection is dose-dependent , and the activation of chloride channel is closely related to the hemolysis induced by puerarin injection.