Sonoporation using microbubbles promotes lipofectamine-mediated siRNA transduction to rat retina.
Sonoporation using microbubbles promotes lipofectamine-mediated siRNA transduction to rat retina.
复制标题
DOI：
10.17305/bjbms.2011.2565
复制
发表时间：
2011-08
影响因子：
3.4
通讯作者：
Xiao-zhi Zheng;Ping Ji;Jianqun Hu
中科院分区：
文献类型：
--
作者：
Xiao-zhi Zheng;Ping Ji;Jianqun Hu
超声靶向微泡破坏（UTMD）已被用于在体外环境中将裸siRNA递送到细胞中。但是UTMD是否可以安全地将裸siRNA递送到体内细胞中仍然是未知的。本工作旨在研究UTMD增强裸siRNA转导（或与Lipofectamine 2000组合）在体内视网膜细胞中的可行性，并比较UTMD和单独超声照射在这种增强作用中的性能。将Cy 3标记的siRNA在不同的Lipofectamine 2000或/和UTMD条件下注射到大鼠眼玻璃体腔内。通过荧光显微镜和流式细胞术评估转导效率。分别通过台盼蓝排斥试验和苏木精-伊红染色评估细胞和组织损伤。Lipofectamine 2000和UTMD组转染细胞的数量和密度明显高于其他组。接受Lipofectamine 2000和单独超声照射的组中的转导细胞数略多于接受Lipofectamine 2000的组。在单独接受UTMD的组中也可以看到Cy 3-siRNA阳性细胞，尽管转导效率极低。各组细胞存活率均大于90%，视网膜结构保存良好。这些结果表明，UTMD具有显著高于单独超声辐照的性能，可以有效地增强Lipofectamine 2000介导的裸siRNA在体内视网膜细胞中的转导，而不对细胞或组织造成任何损伤。该方法可作为治疗眼底疾病的一种新方法。
Ultrasound-targeted microbubble destruction(UTMD) has been utilized to deliver naked siRNA into cells in in vitro settings. But whether UTMD can safely deliver naked siRNA into in vivo cells have remained unknown. This work was performed to investigate the feasibility of UTMD-enhanced naked siRNA transduction (or combined with Lipofectamine 2000) in vivo retinal cells and compare the performance between UTMD and ultrasonic irradiation alone in this enhancing effect. A dose of Cy3-labeled siRNA was injected into the vitreous cavity of rat eyes under the different conditions of Lipofectamine 2000 or/and UTMD. Transduction efficiency was assessed by fluorescence microscopy and flow cytometry. Cell and tissue damage was assessed by trypan blue exclusion test and hematoxylineosin staining, respectively. The quantity and the density of transducted cells in the group received Lipofectamine 2000 and UTMD was far more than that in other groups. The number of transducted cells in the group received Lipofectamine 2000 and ultrasonic irradiation alone was slightly more than that in the group received Lipofectamine 2000. Cy3-siRNA-positive cells can also seen in the group received UTMD alone, although the transduction efficiency is extremely low. Cell viability in each group was more than 90%, and retinal architecture in each group was well preserved. These results indicated that UTMD, with a significantly higher performance than ultrasonic irradiation alone, can effectively enhance the Lipofectamine 2000-mediated naked siRNA transduction in vivo reinal cells without any cell or tissue damage. This method can serve as a novel approach to treat the diseases of eye ground.