Cardiovascular Center, Departments of Internal Medicine and Medical Research, Mackay Memorial Hospital, Taipei City, Taiwan.
Cardiovascular Center, Departments of Internal Medicine and Medical Research, Mackay Memorial Hospital, Taipei City, Taiwan; Mackay Medical College, New Taipei City, Taiwan.
Ultrasound Med Biol. 2019 Sep;45(9):2434-2443. doi: 10.1016/j.ultrasmedbio.2019.05.009. Epub 2019 Jun 24.
We investigated whether ultrasonic microbubble transfection (UMT) would enhance the transfection of large-sized luciferase plasmids (5.6, 9.2 and 33 kb) and biological impacts. Porcine venous blood endothelial progenitor cells (EPCs) were cultured in a medium containing plasmid DNA (pDNA) of different sizes followed by UMT and functional assays. Real-time polymerase chain reaction was conducted to investigate the effects of transfection of pDNA on multiple molecules central to endothelial function. The results indicated enhanced luciferase expression after UMT but the enhancement declined with increase in the size of the plasmid. UMT of pDNAs sized 5.6 and 9.2 kb into EPCs led to significant enhancement of proliferation. The interleukin-6 (IL-6) secreted from UMT of EPCs also increased in the 5.6- and 9.2-kb pDNA groups. Treatment of the transfected EPCs with anti-IL-6 antibody neutralized the proliferation. In conclusion, UMT of pDNAs sized 5.6 and 9.2 kb into EPCs increased the secretion of IL-6, which in turn enhanced cell proliferation.
我们研究了超声微泡转染（UMT）是否会增强大型荧光素酶质粒（5.6、9.2 和 33 kb）的转染效率和生物学影响。将不同大小的质粒 DNA（pDNA）培养在含有猪静脉血内皮祖细胞（EPCs）的培养基中，随后进行 UMT 和功能检测。通过实时聚合酶链反应研究了转染 pDNA 对内皮功能相关多个分子的影响。结果表明，UMT 后荧光素酶表达增强，但随着质粒大小的增加，增强效果下降。将 5.6 和 9.2 kb 大小的 pDNA 转染到 EPCs 中可显著增强细胞增殖。转染的 EPCs 中白细胞介素-6（IL-6）的分泌也在 5.6 和 9.2 kb pDNA 组中增加。用抗 IL-6 抗体处理转染的 EPCs 可中和细胞增殖。总之，将 5.6 和 9.2 kb 大小的 pDNA 转染到 EPCs 中会增加 IL-6 的分泌，进而增强细胞增殖。
