葛根素PEG-PE纳米胶束的制备及降低红细胞溶血的初步研究

Preparation of Puerarin-loaded PEG-PE Nano-micelle and Its Reducing Hemolysis Effect

关键词:  葛根素PEG-PE纳米胶束  红细胞溶血  红细胞脆性  红细胞膜流动性

Key words:  puerarin-loaded PEG-PE nano-micelle  hemolysis  erythrocyte osmotic fragility  membrane fluidity of akaryocyte

摘要:

目的 采集葡萄糖-6-磷酸脱氢酶(glucose-6-phosphate ehydrogenase deficiency，G6PD)缺陷的动物模型血液，评价葛根素PEG-PE纳米胶束降低红细胞溶血的作用。方法 1%乙酰苯肼皮下注射大鼠制备G6PD缺陷的动物模型，取其红细胞制成混悬液，比较评价葛根素和葛根素PEG-PE纳米胶束的溶血作用、红细胞脆性及红细胞膜流动性。结果 葛根素出现了明显的红细胞溶血，且随着浓度变化和孵育时间延长具有增加的趋势，溶血率高达38%，而葛根素PEG-PE纳米胶束未见明显溶血；另外，红细胞脆性实验结果表明，葛根素PEG-PE纳米胶束对红细胞膜的作用非常轻微，H_50%为(42.1±1.1)%，显著小于葛根素的该数值(53.1±1.2)%，红细胞膜流动性结果表明，葛根素PEG-PE纳米胶束对红细胞膜的流动性影响非常轻微。结论 葛根素PEG-PE纳米胶束良好的血液相容性，为该药物的体内研究和进一步新型药物制剂开发奠定基础。

Abstract:

OBJECTIVE To explore and analyze the reducing hemolytic effects of puerarin-loaded PEG-PE nano-micelle via the glucose-6-phosphate dehydrogenase (G6PD)-deficient rats blood. METHODS The rat model with G6PD-deficiency was established via injecting 1% acetylphenyl-hydrazine. Then the G6PD-deficient erythrocyte suspension obtained from this rat model was used to evaluate the hemolytic effects of PUE and puerarin-loaded PEG-PE nano-micelle via hemolytic activity, erythrocyte osmotic fragility assay and membrane fluidity of erythrocytes. RESULTS PUE could cause a serious hemolysis to the erythrocyte suspension with the increase of drug concentration and the prolongation of drug incubation time, the hemolytic rate of PUE was up to 38%, while PEG-PEN revealed no significant hemolysis. Additionally, the result of erythrocyte osmotic fragility indicated that puerarin-loaded PEG-PE nano-micelle exerted a slight effect on the erythrocyte membranes, its H_50% was (42.1±1.1)%, much less than that (53.1±1.2)% of PUE. Erythrocyte membrane fluidity indicated that the effect of puerarin-loaded PEG-PE nano-micelle on erythrocyte membrane fluidity was very slight. CONCLUSION The good blood compatibility of puerarin-loaded PEG-PE nano-micelle makes it a favorable candidate for in vivo tests and puerarin-loaded PEG-PE nano-micelle can also provide the useful insight for the further formulation development as an innovative drug.