共载阿霉素与小干扰RNA的脂质介孔硅纳米粒的制备表征及抗多药耐药肿瘤细胞研究  

Preparation and characterization of doxorubicin and siRNA co-loaded CLMSNs and study on antimultidrug-resistant tumor cells

作　　者：张梦玮 杨硕晔[1,2] 杨亚南 王振威 屈凌波 ZHANG Mengwei;YANG Shuoye;YANG Yanan;WANG Zhenwei;QU Lingbo(College of Biological Engineering,Henan University of Technology,Zhengzhou 450001,China;Zhengzhou Key Lab of Biomedical Functional Molecules,Zhengzhou 450001,China;College of Chemistry and Molecular Engineering,Zhengzhou University,Zhengzhou 450001,China)

机构地区：[1]河南工业大学生物工程学院,郑州450001 [2]郑州市生物医药功能分子重点实验室,郑州450001 [3]郑州大学化学与分子工程学院,郑州450001

出　　处：《中国药房》2022年第23期2880-2885,共6页China Pharmacy

基　　金：河南省高等学校青年骨干教师培养计划(No.2020GGJS090);河南工业大学青年骨干教师培育计划(No.21420075)。

摘　　要：目的制备共载阿霉素(DOX)与小干扰RNA(siRNA)的脂质介孔硅纳米粒(CLMSNs-SS-NH_(2)@DOX/siRNA),并对其进行表征及抗多药耐药肿瘤细胞研究。方法先以介孔硅纳米粒(MSNs)为基础制备MSNs-SS-NH_(2)@DOX,并以阳离子脂质体(CLs)对其包覆制得CLMSNs-SS-NH_(2)@DOX,进一步负载siRNA即得CLMSNs-SS-NH_(2)@DOX/siRNA。测定该制剂粒径及Zeta电位,观察其微观形态并进行差示扫描量热分析、X射线衍射分析、红外光谱分析、物理吸附分析;测定该制剂在不同pH条件下(pH5.0、pH7.4)及不同谷胱甘肽浓度下(0、2、5、10 mmol/L)DOX的体外释放度;考察该制剂对耐DOX型乳腺癌细胞MCF-7/ADR摄取、迁移、凋亡、周期及P-糖蛋白(P-gp)表达的影响。结果CLMSNs-SS-NH_(2)@DOX/siRNA结构清晰,表面脂膜层明显,粒径为(197.63±3.75)nm,Zeta电位为(20.64±0.98)m V,理化性质良好。体外释放结果显示,CLMSNs-SS-NH_(2)@DOX/siRNA具有良好的p H/还原双重响应释药特性。细胞实验结果显示,经CLMSNs-SS-NH_(2)@DOX/siRNA干预后,MCF-7/ADR细胞的药物摄取显著增强,迁移率和P-gp表达水平均显著降低,总凋亡比例和G_(0)/G_(1)期所占比例均显著升高(P<0.05)。结论本研究成功制备同时负载DOX和siRNA的CLMSNs-SS-NH_(2)@DOX/siRNA;该制剂理化性质良好,具有pH/还原双重响应释药特性,且可通过下调P-gp表达逆转MCF-7/ADR细胞多药耐药。OBJECTIVE To prepare lipid-coated mesoporous silica nanoparticles(CLMSNs)co-loaded with doxorubicin(DOX)and siRNA(CLMSNs-SS-NH_(2)@DOX/siRNA),and to characterize it and study anti-multidrug-resistant tumor cells.METHODS MSNs-SS-NH_(2)@DOX was prepared on the basis of mesoporous silica(MSNs),covered with cationic liposomes(CLs)to synthesize CLMSNs-SS-NH_(2)@DOX,and then obtain CLMSNs-SS-NH_(2)@DOX/siRNA by co-loading with siRNA.The particle size and Zeta potential of the preparation were determined,and its micromorphology was observed;differential scanning calorimetry,X-ray diffraction,infrared spectroscopy and physical adsorption analysis were conducted.The in vitro release of DOX from the preparation was determined under different pH conditions(pH5.0,pH7.4)and different glutathione concentrations(0,2,5,10 mmol/L).The effects of this preparation on the uptake,migration,apoptosis,cycle and P-glycoprotein(P-gp)expression of MCF-7/ADR in DOX-resistant breast cancer cells were investigated.RESULTS CLMSNs-SS-NH_(2)@DOX/siRNA had a clear core-shell structure,obvious lipid membrane layer,particle size of(197.63±3.75)nm,Zeta potential of(20.64±0.98)mV,and with good physical and chemical properties.In vitro release results showed that CLMSNs-SS-NH_(2)@DOX/siRNA possessed good pH/reduction double-response.The results of cell experiment showed that after intervened with CLMSNs-SS-NH_(2)@DOX/siRNA,the fluorescence intensity of MCF-7/ADR cells was significantly enhanced,the migration rate and P-gp expression level were significantly reduced,while total proportion of apoptosis and that of G_(0)/G_(1) phase were significantly increased(P<0.05).CONCLUSIONS In this study,DOX and siRNA co-loaded CLMSNs-SS-NH_(2)@DOX/siRNA is prepared successfully,which has good physical and chemical properties,pH/reduction double-response properties.It can reverse the multidrug resistance of MCF-7/ADR cells by downregulation of P-gp expression.

关 键 词：介孔硅 小干扰RNA 阿霉素 抗肿瘤 多药耐药 

分 类 号：R944.9[医药卫生—药剂学] R979.1[医药卫生—药学]