氯通道在葛根素注射液致溶血反应中的作用

目的：观察阻断和激活氯通道对葛根素注射液致家兔溶血反应的影响，探讨氯通道在葛根素注射液溶血反应中的作用。方法：将不同浓度的葛根素注射液（0.75、1.5、3、6、12 mg/ml）与家兔红细胞悬液共孵育6 h，使用细胞图像记录系统观测葛根素注射液是否诱导红细胞溶血，酶标仪、流式细胞仪检测红细胞溶血率，并观测激活和阻断氯通道对葛根素注射液溶血作用的影响。结果：葛根素注射液可引起家兔红细胞体外溶血，在所观察的1.5 mg/ml~12 mg/ml范围内，溶血效应呈浓度依赖性增强（n=3，P<0.01）。氯通道阻断剂Tamoxifen （20 μmol/L）、ATP （10 mmol/L）可有效抑制葛根素注射液的溶血作用（n=3~5，P<0.01）；而使用低浓度ATP （50 μmol/L）激活氯通道，则显著增强葛根素注射液的溶血作用（n=4，P<0.01）。结论：葛根素注射液的体外溶血效应呈浓度依赖性，氯通道激活与葛根素注射液诱导的溶血反应密切相关。

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 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.