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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

25卷

期数:

2019年03期

页码:

215-219

栏目:

基础医学

出版日期:

2019-05-20

文章信息/Info

Title:

Effect of hexoses on Pip5e-PMO mediated exon-skipping in the Duchenne muscular dystrophy mouse model

文章编号:

1006-8147(2019)03-0215-05

作者:

宁含含1; 顾 犇1; 韩 刚2; 林曹瑞1; 尹海芳1

（1.天津医科大学基础医学院细胞生物学系，天津300070；2.天津医科大学医学检验学院，天津300203）

Author(s):

NING Han-han1;  GU Ben1;  HAN Gang2;  LIN Cao-rui1;  YIN Hai-fang1

（1. Department of Cell Biology, School of Basic Medical Sciences, Tianjin Medical University, Tianjin 300070, China; 2.School of Medical Laboratory, Tianjin Medical University, Tianjin 300203,China）

关键词:

杜兴肌肉萎缩症; 外显子跳读; 己糖;  肽修饰PMO;  抗肌萎缩蛋白

Keywords:

DMD;  exon skipping;  hexose;  Pip5e-PMO;  dystrophin

分类号:

Q291

DOI:

-

文献标志码:

A

摘要:

目的：在杜兴肌肉萎缩症模型上探究己糖混合物对核酸药物Pip5e-PMO介导的外显子跳读活性的促进作用。方法：选择成年mdx小鼠为测试动物模型，通过局部肌肉和系统尾静脉注射，比较Pip5e-PMO与己糖和生理盐水联用的作用效果，利用免疫荧光（IHC）和蛋白印迹法（Western blot）检测dystrophin阳性肌纤维的数量和分布及dystrophin蛋白的表达水平；利用逆转录-聚合酶链式反应（RT-PCR）检测外显子跳读。结果：在局部肌肉给药试验中，己糖能够促进Pip5e-PMO诱导dystrophin蛋白表达水平（P<0.05）。在系统试验中，对己糖和生理盐水组比较发现：按15 mg/kg的剂量单次系统尾静脉注射Pip5e-PMO，骨骼肌中dystrophin蛋白表达恢复至正常水平的50%；在心脏中，也检测到明显的dystrophin恢复表达和外显子跳读。但两组无显著性差别（P>0.05）。结论： Pip5e-PMO可诱导有效的外显子跳读和dystrophin蛋白表达，但己糖对其促进效果有限。

Abstract:

Objective: To explore the potential of hexose(the mixture of glucose and fructose -GF) in enhancing the delivery of peptide-modified PMO (Pip5e-PMO) in the Duchenne Muscular Dystrophy(DMD) mouse model (mdx mouse). Methods: Pip5e-PMO in GF or saline was intramuscularly or intravenously administered into adult mdx mice. Immunohistochemistry(IHC) and Western blot were used to measure the distribution and expression of dystrophin. Reverse transcription-Polymerase chain reaction (RT-PCR) was utilized to examine the level of exon skipping. Results: Local intramuscular results showed that GF enhanced the activities of Pip5e-PMO demonstrated by the increased number of dystrophin-positive fibers and level of dystrophin expression（P<0.05）. Although single intravenous injection of 15mg/kg Pip5e-PMO induced substantial number of dystrophin-positive fibers and therapeutic levels of dystrophin restoration in peripheral muscles and heart of mdx mice treated with either GF or saline, the difference between GF and saline groups was not significant（P>0.05）. Conclusion: GF could enhance the activities of Pip5e-PMO intramuscularly but may fail to have any impact on dystrophin-deficient mdx mice.

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备注/Memo

备注/Memo:

基金项目 天津市教委科研计划项目基础理论研究项目（2016YD10）
作者简介 宁含含（1993-），女，硕士在读，研究方向：医学细胞生物学；
通信作者：尹海芳，E-mail:haifangyin@tmu.edu.cn。

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更新日期/Last Update: 2019-07-03