Mg2Al-Cl-LDH载体转染细胞的研究

Transfection of Mg_2Al-Cl-LDH nanoparticles as gene carriers in cells

为探讨Mg2Al-Cl-LDH颗粒作为基因转染载体的可能性以及最优的转染体系和转染时间,为后续的基因治疗、创面修复、转基因技术等试验打下基础,自制Mg2Al-Cl-LDH纳米颗粒,与增强型荧光蛋白质基因eGFP结合后,转染入细胞,观察转染后细胞表达的绿色荧光蛋白质,计算转染率。结果表明Mg2Al-Cl-LDH纳米颗粒直径在100 nm左右,结构为六边形。200μg/ml的纳米颗粒浓度下细胞活性为87%,达到500μg/ml时细胞活性快速降至44%;DNA与LDH的质量比为4/50时转染率达到11.17%,质量比为10/50时,转染效率为3.93%;DNA与LDH作用时间在15 min时转染率为6.93%,30 min时反而下降至4.8%;转染细胞培养24 h时转染率达到17.4%,72 h后下降至3.7%。纳米颗粒的分子较小,比较容易被细胞膜胞吞,并且Mg2Al-Cl-LDH纳米颗粒能有效地与eGFP质粒相结合;在DNA与LDH质量比4/50的情况下,结合能力最高;最佳结合时间为15 min;转染时间在48 h时效果最好。HEK 293T细胞对Mg2Al-Cl-LDH纳米颗粒的最优耐受浓度为200μg/ml;同样条件下C2C12细胞的转染效率比HEK 293T细胞的转染效率高。

In order to figure out the possibility of Mg2Al-Cl-LDH nanoparticles as gene carriers and determine the optimal transfeetion condition and time, so as to lay the foundation for gene therapy, wound repair, and transgenic technol- ogy, 10-nm diameter Mg2Al-Cl-LDH nanoparticles with hexagonal structure were prepaned. Assembled with enhanced eG- FP fluorescent protein gene, the particles were transfected into cells, and the expression of green fluorescent protein and transfection rate were assayed. The results revealed that cell activity was 87% under the nanoparticles concentration of 200μg/m and rapidly decreased to 44% when nanoparticles concentration reached 500 μg/m. The transfection rate was 11.17% under 4/50 mass ratio of DNA to LDH, and droped to 3.93% when the mass ratio was 10/50. The transfection rate was 6. 93% after DNA and LDH func- tioned for 15 min, and decreased to 4. 8% for 30 min. Trausfection rate was 17.4% after cell culture for 24 h and decreased to 3.7% after 72 h. Nanoparticle molecule was small and cell membrane endocytosis-prone. Mg2Al-Cl-LDH nanoparticles could be efficiently combined with eGFP plasmid. The highest transfection capacity appeared in the sys- tem of 4/50 mass ratio of DNA to LDH, 15 min of DNA-LDI-I binding, 48 h of cell culturing, and nanoparticle concentra- tion of 200 mg/L. Under the same condition, transfection efficiency in C2 Cl2 cells was higher than that of HEK 293T cells.