首页> 中文期刊>华西口腔医学杂志 >MT01/PEN复合物对人成骨样细胞MG63表达骨保护蛋白和核因子κB受体活化因子配体的影响

MT01/PEN复合物对人成骨样细胞MG63表达骨保护蛋白和核因子κB受体活化因子配体的影响

     

摘要

Objective This study aims to synthesize MT01 (a kind of oligodeoxynucleotides) and N-isopropylacrylamidemodified polyethylenimines (PEN) complexes (MT01/PEN) as well as to investigate the effect of the complexes on the expression of osteoprotegerin (OPG) and the receptor activator of nuclear factor κB ligand (RANKL) in the human osteoblast-like cell line MG63. Methods MG63 cells were transfected by MT01/PEN complexes formed with three different mass ratios (1∶2, 1∶4, 1∶6) of MT01 to PEN. MT01 and MT01-s were used as positive control. Enzyme-linked immunosorbent assay and real-time polymerase chain reaction were performed to estimate the amount of OPG and RANKL released into the culture media and in MG63 at 24, 48, 72 h. Results MG63 responded to the MT01/PEN complexes by significantly upregulating the OPG on the protein and mRNA levels (P<0.05). The protein and mRNA levels of RANKL were lower in most of the groups with complexes, and the OPG/RANKL ratio were higher (P<0.05). MG63 were affected by the MT01/PEN complexes with different mass ratios, particularly when the ratio was 1∶6. Conclusion MT01 can enhance the promotion of ossification by establishing the delivery system with PEN.%目的:   应用聚乙烯亚胺阳离子聚合物(PEN)载体装载寡脱氧核苷酸MT01,制备MT01/PEN复合物,检测该复合物对人成骨样细胞MG63表达骨保护蛋白(OPG)和核因子κB受体活化因子配体(RANKL)的影响。方法 制备3种不同配比的MT01/PEN(质量比分别为1∶2、1∶4、1∶6)复合物,以全硫代化修饰MT01(MT01-s)和未修饰的MT01作阳性对照,分别转染MG63细胞。采用酶联免疫吸附测定法和real-time聚合酶链反应法分别检测培养24、48、72 h时各组上清液及细胞内OPG和RANKL的表达水平。结果   经MT01/PEN复合物转染后,上清液及细胞内OPG表达水平均升高(P<0.05);多数组中RANKL表达水平降低,而OPG/RANKL比值呈升高趋势(P<0.05);不同质量配比的MT01/PEN均对MG63细胞的成骨具有影响,其中质量比为1∶6时作用最明显。结论   应用PEN作为基因载体装载MT01可增强MT01对MG63细胞的促成骨作用。

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