活性氧响应负载环孢菌素A纳米粒对缺氧/复氧损伤心肌细胞的保护作用

Protective effect of reactive oxygen species responsive cyclosporin A nanoparticles on hypoxia/reoxygenation injured cardiomyocytes

目的制备负载环孢菌素A(CsA)的聚5,5-二甲基-4,6-二硫基-壬二酸丙二醇酯(PTK)纳米粒(CsA@PTK),研究其活性氧(ROS)特性及对H9c2心肌细胞缺氧/复氧损伤的保护作用。方法采用乳化溶剂挥发法制备CsA@PTK,单因素实验考察超声功率、油水相比例、聚乙烯醇(PVA)质量浓度及CsA投药量对CsA@PTK粒径的影响;检测CsA@PTK的粒径和Zeta电位,用透射电镜(TEM)观察其形态;用透析法评价CsA@PTK的体外释药特性;用MTT法测定CsA@PTK对H9c2心肌细胞缺氧/复氧损伤的保护作用。结果CsA@PTK的平均粒径为200±3 nm,多分散系数(PDI)为0.096±0.022,Zeta电位为-13.41±1.55 mV,载药量为5.16%±0.19%,TEM证实CsA@PTK的形态为球形;CsA@PTK体外释药速率呈ROS浓度依赖性;MTT结果证实,CsA@PTK能提高缺氧/复氧损伤H9c2心肌细胞的存活率。结论CsA@PTK的体外释药性能良好,具有ROS响应特性,对H9c2心肌细胞缺氧/复氧损伤具有保护作用。

Objective To prepare cyclosporin A(CsA)nanoparticles by using poly 5,5-dimethyl-4,6-dithio-propylene azelate(PTK)(CsA@PTK)and to evaluate their response to reactive oxygen species(ROS)and protective effect on hypoxia/reoxygenation injury of H9c2 cardiomyocytes in vitro.Methods CsA@PTK was prepared by emulsification-solvent evaporation method.The effects of ultrasound power,volume ratio of aqueous phase to oil phase,concentration of polyvinyl alcohol(PVA)and dosage of CsA on particle size were evaluated by single factor experiments.The particle size and Zeta potential of nanoparticles were measured by laser particle size analyzer,the morphology was observed by transmission electron microscopy(TEM).The drug release property of CsA@PTK was evaluated by dialysis method.And the protective effect of CsA@PTK on hypoxia/reoxygenation injury of H9c2 cardiomyocytes was determined by MTT method.Results The average size of CsA@PTK was 200±3 nm,PDI was 0.096±0.022,Zeta potential was-13.41±1.55 mV,and drug loading was 5.16%±0.19%.TEM confirmed that CsA@PTK was spherical.The In vitro release profile of CsA@PTK was ROS concentration dependent.MTT results confirmed that CsA@PTK increased the viability of hypoxia/reoxygenation injured H9c2 cardiomyocytes.Conclusion The release of CsA from CsA@PTK in vitro showed an ROS dependent manner,and can protect H9c2 cardiomyocytes from hypoxia/reoxygenation injury.