【作者】 张毅；

【导师】 章东；

【作者基本信息】 南京大学 ， 声学， 2018， 硕士

【摘要】 超声作用下微泡的声孔效应是一种极具潜力的介导药物输运/基因转染的新型手段,相较于其他方法,微泡声孔效应可以在有效提高靶向药物转染效率的同时减小对周围正常组织的副作用。但是其作用机理尚不明确,尤其是细胞周期对超声联合微泡介导药物输运的影响仍有待研究。在本论文的工作中,我们分别使用洛伐他汀、含羞草素、诺考达唑将HeLa细胞同步于G1、S、G2/M期,并使用原子力显微镜(AFM)观测其形貌及弹性模量。之后使用微泡介导的超声作用于不同周期的细胞上,并使用流式细胞仪分析其药物转染效率及活性。实验发现不同时期细胞体积顺序为G1<S<G2/M,杨氏模量顺序为G1>G2/M>S。且受细胞体积的影响,同步于各时期的细胞药物转染效率顺序为G1<S<G2/M。受细胞膜杨氏模量的影响,随惯性空化剂量的增加,不同时期药物转染效率变化量及活性变化率的顺序均为G1<G2/M<S。本文的研究结果对于深入理解超声联合微泡介导基因/药物转染的作用机理提供了重要的实验依据,为进一步控制和优化微泡声穿孔效应在靶向药物输运/基因转染领域的应用奠定了坚实基础。更多还原

【Abstract】 Microbubble-mediated sonoporation has a great potential for targeted drug/gene-transfection.Compared with other methods,it can not only effectively improve the efficiency of targeted drug transfection,but also reduce the side effect on normal tissues nearby.However,the mechanism underlying microbubble-facilitated gene/drug transfection is still under clear,and the influence cell-cycle-phases on microbubble-mediated sonoporation remains to be elucidated.In this study,HeLa cells were synchronized to G1,S and G2/M phases using lovastatin,mimosine and nocodazole,respectively.Their morphology and stiffness were evaluated with atomic force microscopy(AFM).Then,ultrasound sonication was applied to synchronized cells with the presence of microbubbles,and the drug transfection efficiency and cell viability were analyzed by the flow cytometry.The result showed that,the orders of cell volume and Young’s modulus were G1<S<G2/M and G1>G2/M>S,respectively.Due to the effect of cell volume,the drug transfection efficiency follows the sequence that G1<S<G2/M.It was also noticed that,under the influence of Young’s Modulus,the order of change of drug transfection efficiency and change rate of cell viability for three phases was G1<G2/M<S with the increase of ICD.In this paper,we have found the relations between drug transfection efficiency for different cell-cycle-phases,which is important for better understanding the mechanisms underlying microbubble-facilitated gene/drug transfection.The results of the current work might be beneficial for controlling and optimizing the applications of microbubble sonoporation in the field of therapeutic ultrasound.更多还原