包被siRNA的叶酸改性聚乙二醇1000维生素E琥珀酸酯纳米材料的制备及其表征  

Preparation and characterization of folic acid modified D-α-tocopheryl polyethylene glycol 1000 succinate nanomaterials encapsulated with siRNA

作　　者：朱嫚嫚 程勇 饶鹏 张贵阳 刘昊[3] 肖雷[4] 刘加涛 Zhu Manman;Cheng Yong;Rao Peng;Zhang Guiyang;Liu Hao;Xiao Lei;Liu Jiatao(School of Pharmacy,Anhui Medical University,Hefei 230032;School of Basic Medicine,Anhui Medical University,Hefei 230032;Dept of Oncology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Pharmacy,The First Affiliated Hospital of Anhui Medical University,Hefei 230022)

机构地区：[1]安徽医科大学药学院,合肥230032 [2]安徽医科大学基础医学院,合肥230032 [3]安徽医科大学第一附属医院肿瘤内科,合肥230022 [4]安徽医科大学第一附属医院药剂科,合肥230022

出　　处：《安徽医科大学学报》2023年第11期1865-1871,共7页Acta Universitatis Medicinalis Anhui

基　　金：安徽省自然科学基金(编号:2008085MH257);安徽高校科学研究项目(编号:KJ2020A0177)。

摘　　要：目的构建可以稳定负载并有效释放siRNA的叶酸改性聚乙二醇1000维生素E琥珀酸酯(FA-TPGS)纳米材料并观测其对小鼠巨噬细胞RAW264.7细胞毒性和剪切型X-框结合蛋白1(XBP1s)表达水平的影响。方法将FA-TPGS与罗丹明B(RhB)标记的XBP1 siRNA按照5∶1比例混合均匀得到包被XBP1 siRNA的纳米复合物(FT@XBP1)。通过透射电镜、动态光散射、紫外可见光谱和荧光光谱分析对FT@XBP1表征,同时计算XBP1 siRNA从FT@XBP1纳米载体中释放的药量。应用扫描电镜(SEM)、CCK-8以及流式细胞术检测FT@XBP1的细胞毒性,并应用Western blot法检测FT@XBP1对小鼠巨噬细胞RAW264.7中XBP1s的抑制作用。结果FA-TPGS与siRNA具有良好的结合作用,其中FT@XBP1的平均粒径为(200±20)nm。相对释放结果提示,酸性环境(pH 5.0)有利于siRNA从FT@XBP1中释放。CCK-8和凋亡实验均显示,FT@XBP1对RAW264.7细胞的增殖和凋亡影响较小,且FT@XBP1处理可显著抑制RAW264.7中XBP1s蛋白的表达(P<0.001)。结论叶酸修饰的TPGS纳米载体可有效包载XBP1 siRNA,抑制巨噬细胞XBP1s表达且细胞相容性良好。Objective To construct folate modified D-α-tocopheryl polyethylene glycol 1000 succinate(TPGS)nanomaterials(FA-TPGS),which can stably load and effectively release siRNA,and to observe the effects of nanoparticles on the cytotoxicity and spliced X-box binding protein 1(XBP1s)of mouse leukemia cells of monocyte macrophage(RAW264.7).Methods Mixed FA-TPGS and rhodamine B(RhB)labeled XBP1 siRNA solution in a proportion of 5∶1 and obtained the nano-complex coated with XBP1 siRNA(FT@XBP1).FT@XBP1 nanocarriers were characterized by transmission electron microscope,dynamic light scattering,ultraviolet visible spectrum analysis and/or fluorescence analysis.And the release of siRNA from FA-TPGS nano-carriers was calculated simultaneously.The cell cytotoxicity of FT@XBP1 nanomaterials were detected by scanning electron microscopy(SEM),CCK-8 and flow cytometry.And the inhibited effect of XBP1s of RAW264.7 cells was checked by Western blot.Results FA modified TPGS could effectively bind XBP1 siRNA.And the average particle size of FT@XBP1 nanocarriers were(200±20)nm.The relative release assays showed that acidic environments(pH 5.0)was beneficial for siRNA to release from [e-mail] CCK-8 and apoptosis assay showed that the effects of FT@XBP1 on the proliferation and apoptosis of RAW264.7 cells were relatively small,and FT@XBP1 could significantly inhibit the expression of XBP1s protein in RAW264.7(P<0.001).Conclusion TPGS nanoparticles modified with folic acid can effectively encapsulate XBP1 siRNA and inhibit XBP1s expression of RAW264.7 cells with good cellular compatibility.

关 键 词：叶酸 聚乙二醇1000维生素E琥珀酸酯 SIRNA 巨噬细胞 X-框结合蛋白1 

分 类 号：R318.08[医药卫生—生物医学工程]