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| 靶向声敏纳泡的制备及其声动力效应观察 |
| 陈粤瑛,王浩,胡玉刚,黄鑫,陈金玲,邓倾,曹省,潘桔红,周青 |
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| (武汉大学人民医院超声影像科) |
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| 摘要: |
| 目的 制备以动脉粥样硬化斑块A型清道夫受体（SR-A）为靶点的载二氢卟吩e6（Ce6）超声纳泡（NB），探讨其提高声敏剂的靶向性及声动力效应的能力。
方法 以薄膜水化法制备载Ce6和SR-A靶向多肽PP1的纳泡（Ce6-PP1 NB），并检测其理化性质和细胞毒性。将小鼠单核巨噬细胞白血病细胞（RAW264.7细胞）分别与Ce6、Ce6-PP1 NB共孵，采用共聚焦激光显微镜观察细胞内Ce6荧光强度以评估Ce6-PP1 NB的靶向性。将RAW264.7细胞分成4组：对照组、低强度脉冲超声（LIPUS）组、Ce6+LIPUS组和Ce6-PP1 NB+LIPUS组，采用活性氧荧光探针（DCFH-DA）法、流式细胞术和酶联免疫吸附测定（ELISA）法评估Ce6-PP1 NB的声敏性和声动力效应。
结果 制备的Ce6-PP1 NB呈圆形，形态规则，分散均匀，粒径为（504.25±149.13）nm，zeta电位为（-8.67±0.78）mV，Ce6包封率为（85.63±4.39）%，PP1连接率为（92.78±4.57）%。共聚焦激光显微镜下示Ce6-PP1 NB组细胞内Ce6红色荧光明显高于Ce6组。与Ce6+LIPUS组比较，Ce6-PP1 NB+LIPUS组细胞内出现明显绿色荧光，M1型巨噬细胞占比减少，M2型巨噬细胞占比增加，细胞分泌肿瘤坏死因子-α（TNF-α）、白细胞介素-6（IL-6）减少，而分泌白细胞介素-10（IL-10）增加，提示Ce6-PP1 NB联合LIPUS的声动力效果优于Ce6联合LIPUS。
结论 Ce6-PP1 NB有助于提高声敏剂在动脉粥样硬化斑块的聚集和声动力效应。 |
| 关键词: 声动力治疗 动脉粥样硬化 巨噬细胞极化 纳泡 |
| DOI： |
| 投稿时间：2022-08-15修订日期：2022-11-15 |
| 基金项目:国家自然科学基金项目（面上项目，重点项目，重大项目） |
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| Enhanced Sonodynamic Therapy with Atherosclerosis-targeted Sonosensitive Nanobubbles |
| CHEN Yueying,WANG Hao,HU Yugang,HUANG Xin,CHEN Jinling,DENG Qing,CAO Sheng,PAN Juhong,ZHOU Qing |
| (Department of Ultrasonography,Renmin Hospital of Wuhan University) |
| Abstract: |
| Objective To prepare chlorine e6 （Ce6）-loaded ultrasound nanobubbles （NB）targeting atherosclerotic plaque scavenger receptor-A （SR-A），and to explore the effect to enhance the targetable ability and sonodynamic efficacy of sonosensitizers.
Methods Nanobubbles （Ce6-PP1 NB） with Ce6 and SR-A-targeted peptides PP1 were prepared by the film hydration method，then their physicochemical properties and cytocompatibility were assessed. Mouse leukemia cells of monocyte macrophage （RAW264.7 cells） were co-incubated with Ce6 and Ce6-PP1 NB，respectively，then the intracellular fluorescence of Ce6 was observed by confocal laser scanning microscope （CLSM） to evaluate the targetable ability of Ce6-PP1 NB.RAW264.7 cells were divided into four groups：control， low-intensity pulsed ultrasound （LIPUS），Ce6+LIPUS and Ce6-PP1 NB+LIPUS.Next，the sonosensitivity and sonodynamic efficacy of Ce6-PP1 NB were evaluated by DCFH-DA staining，flow cytometry，and ELISA assays.
Results Ce6-PP1 NB presented a regular spherical shape and homogenized dispersion with the average particle size of （504.25±149.13）nm，the zeta potential of （-8.67±0.78）mV，Ce6 encapsulation efficiency of （85.63±4.39）% and PP1 loaded capacity of （92.78%±4.57）%.CLSM showed more significant intracellular red fluorescence of Ce6 in the Ce6-PP1 NB group than in the Ce6 group.Compared to cells in Ce6+LIPUS，cells in Ce6-PP1 NB +LIPUS manifested distinct intracellular green fluorescence and secreted less tumor necrosis factor-α （TNF-α） and interleukin-6 （IL-6） but more interleukin-10 （IL-10），with the decreasing M1 phenotype while the increasing M2 phenotype，suggesting that the sonodynamic efficacy of Ce6-PP1 NB+LIPUS was superior to that of Ce6+LIPUS.
Conclusion The successfully constructed Ce6-PP1 NB is expected to improve the targeting ability and sonodynamic efficacy of sonosensitizers in atherosclerotic plaque. |
| Key words: Sonodynamic therapy Atherosclerosis Macrophage polarization Nanobubbles |
