华西口腔医学杂志

• 基础研究 • 上一篇    下一篇

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

崔野1,2  郑义3  申玉芹3  侯旭1  娄译心1  孙新华1   

  1. 1.吉林大学口腔医院正畸科;2.吉林省牙发育及颌骨重塑与再生重点实验室;
    3.吉林大学口腔医院牙周病科,长春 130021
  • 出版日期:2016-02-01 发布日期:2016-02-01
  • 通讯作者: 孙新华,教授,学士,E-mail:xinhuasun8@163.com
  • 作者简介:崔野,硕士,E-mail:279914375@qq.com
  • 基金资助:

    白求恩前沿交叉学科创新基金(2013108031)

Effect of MT01/PEN complexes on the expression of osteoprotegerin and receptor activator of nuclear factor κB ligand in human osteoblast-like cell line MG63

Cui Ye1,2, Zheng Yi3, Shen Yuqin3, Hou Xu1, Lou Yixin1, Sun Xinhua1   

  1. 1. Dept. of Orthodontics, School and Hospital of Stomatology, Jilin University, Changchun 130021, China; 2. Jilin Provincial Key Laboratory of Tooth Development and Bone Remodel, Changchun 130021, China; 3. Dept. of Periodontology, School and Hospital of Stomatology, Jilin University, Changchun 130021, China
  • Online:2016-02-01 Published:2016-02-01

摘要:

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。

关键词: 人成骨样细胞MG63,  , 寡脱氧核苷酸MT01, 聚乙烯亚胺阳离子聚合物, 骨保护蛋白, 核因子&kappa, B受体活化因子配体

Abstract:

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.

Key words: human osteoblast-like cell line MG63, oligodeoxynucleotides MT01, N-isopropylacrylamide-modified polyethylenimines, osteoprotegerin, receptor activator of nuclear factor κB ligand