雄黄纳米粒的制备及其靶向Survivin蛋白诱导A549细胞凋亡的研究  

Preparation of Realgar Nanoparticles and Apoptosis of A549 Cells Induced by Targeted Survivin Protein

作　　者：王胜玫 刘炫均 邓桂明[1] 何娅娜[1] WANG Sheng-mei;LIU Xuan-jun;DENG Gui-ming;HE Ya-na(The First Affiliated Hospital of Hunan University of Chinese Medicine,Changsha 410007,China;Xiangya School of Pharmaceutical Sciences,Central South University,Changsha 410013,China)

机构地区：[1]湖南中医药大学第一附属医院,长沙410007 [2]中南大学湘雅药学院,长沙410013

出　　处：《中国药学杂志》2021年第7期553-558,共6页Chinese Pharmaceutical Journal

基　　金：湖南省高层次卫生人才“225”工程项目资助(湘卫函[2019]196号);湖南中医药大学中医学国内一流建设学科项目资助(湘教通[2018]469号);湖南省中医药科研计划项目资助(2016130)。

摘　　要：目的制备雄黄纳米粒并考察其理化性质,研究雄黄纳米粒对人肺癌细胞A549增殖凋亡的影响。方法以聚乙烯吡咯烷酮K30(PVP)为载体,通过生物模板法制备雄黄纳米粒。采用透射电子显微镜(transmission electron microscope, TEM)、红外光谱分析、紫外光谱分析以及粒度分析等方法对所制备的纳米粒进行表征;原子吸收光谱法测定纳米粒中As_(4)S_(4)的载药量和包封率;噻唑蓝(MTT)法考察雄黄纳米粒对A549细胞的体外抑制活性,并计算半数抑制浓度(IC_(50)),Annexin V/PI双染色法检测A549细胞的凋亡,Western blot法检测survivin蛋白的表达。结果所制备的雄黄纳米粒为形态规则、分散均匀的球形粒子,纳米粒的粒径为50.7 nm左右,电位为-23.3 mV,冻干复溶后纳米粒粒径为68.1 nm左右,电位为-21.4 mV,As_(4)S_(4)@PVP NPs的包封率为25.9%,载药量为13.4%,紫外光谱显示As_(4)S_(4)@PVP NPs中出现PVP的特征吸收峰,红外光谱分析显示As_(4)S_(4)@PVP NPs与PVP的红外光谱峰位一致,As_(4)S_(4)@PVP NPs在H_2O、PBS、RPMI-1640完全培养基条件下的粒径48 h内无明显变化,体外结果显示,纳米雄黄可显著抑制A549细胞的增殖,与对照组相比较,As_(4)S_(4)@PVP NPs能有效诱导细胞凋亡,降低survivin蛋白的表达。结论成功制备雄黄纳米粒,所制纳米粒粒径小且稳定,可通过降低survivin的表达,有效诱导肺癌A549细胞发生凋亡。OBJECTIVE To prepare realgar nanoparticles and investigate their physicochemical properties,study the effect of realgar nanoparticles on the proliferation and apoptosis of human lung cancer cell A549.METHODS Using polyvinylpyrrolidone K30(PVP)as a carrier,realgar nanoparticles were prepared by the biological template method.The prepared nanoparticles were characterized by transmission electron microscope(transmission electron microscope,TEM),infrared spectroscopy,ultraviolet spectroscopy and particle size analysis,and the drug loading and entrapment efficiency of As4S4 in the nanoparticles were determined by atomic absorption spectrometry.The inhibitory activity of realgar nanoparticles on A549 cells in vitro was detected by thiazolyl(MTT)method,and the half inhibitory concentration(IC50)was calculated.The apoptosis of A549 cells was detected by Annexin V/PI double staining,and the expression of survivin protein was detected by Western blot.RESULTS The prepared realgar nanoparticles are spherical particles with regular morphology and uniform dispersion.The size of the nanoparticles is about 50.7 nm and the potential is-23.3 mV.After freeze-drying and reconstitution,the size of the nanoparticles is about 68.1 nm and the potential is-21.4 mV,the encapsulation rate of As4S4@PVP NPs is 25.9%,and the drug loading is 13.4%.The ultraviolet spectrum shows that the characteristic absorption peak of PVP appears in As4S4@PVP NPs,and the infrared spectrum analysis shows the infrared spectrum peaks of As4S4@PVP NPs and PVP.The position of As4S4@PVP NPs did not change significantly within 48 h under the conditions of H2O,PBS,and RPMI-1640 complete media.The in vitro results showed that nano realgar can significantly inhibit the proliferation of A549 cells.Compared with the control group,As4S4@PVP NPs can effectively induce cell apoptosis and reduce the expression of survivin protein.CONCLUSION Realgar nanoparticles are successfully prepared,and the size of the nanoparticles is small and stable,which could effectively induce ap

关 键 词：纳米雄黄 聚乙烯吡咯烷酮 细胞凋亡 SURVIVIN 

分 类 号：R944[医药卫生—药剂学] R965[医药卫生—药学]