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Basic study on detecting strategy for drug/gene doping for athlete

运动员药物/基因兴奋剂检测策略基础研究

基本信息

批准号：
21240062
负责人：
TAKEMASA Tohru
金额：
$ 30.04万
依托单位：
University of Tsukuba
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
2009
资助国家：
日本
起止时间：
2009 至 2012
项目状态：
已结题

项目摘要

To investigate the feasibility of developing a method for detecion of gene doping in power-athletes, we devised an experimental model system. Myostatin is a potent negative regulator of skeletal muscle development and growth, and myostatinknockout mice exhibit a double-muscle phenotype. To achieve knockdown, we constructed plasmids expressing short hairpin interfering RNAs (shRNAs) against myostatin. These shRNAs were transfected into C2C12 cultured cells or injected into the tibialis anterior (TA) muscle of adult mice. By performing in vitro and in vivo experiments, we found that some shRNAs effectively reduced the expression of myostatin, and that the TA muscle showed hypertrophy of up to 27.9%. Then, using real- time PCR, we tried to detect the shRNA plasmid in the serum or muscles of mice into which it had been injected. Although we were unable to detect the plasmid in serum samples, it was detectable in the treated muscle at least four weeks after induction. We were also able to detect the plasmid in muscle in the vicinity of the TA. This gene doping model system will be useful for further studies aimed at doping control.

为了研究一种检测运动员基因兴奋剂的方法的可行性，我们设计了一个实验模型系统。肌生长抑制素是骨骼肌发育和生长的一种有效的负调节因子，肌生长抑制素敲除小鼠表现出双肌表型。为了实现敲除，我们构建了表达短发夹干扰rna （shRNAs）的质粒来对抗肌肉生长抑制素。将这些shrna转染到C2C12培养细胞或注射到成年小鼠胫骨前肌（TA）中。通过体外和体内实验，我们发现一些shrna有效地降低了肌生长抑制素的表达，TA肌出现了高达27.9%的肥大。然后，使用实时PCR，我们试图检测shRNA质粒在小鼠血清或肌肉中被注射的情况。虽然我们无法在血清样本中检测到质粒，但在诱导后至少四周的处理肌肉中可以检测到质粒。我们还能够在TA附近的肌肉中检测到质粒。该基因兴奋剂模型系统将为进一步的兴奋剂控制研究提供依据。

项目成果

期刊论文数量（73）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Genome-wide analysis of sense-antisense transcripts, in "Non-coding RNAs and Epigenetic Regulation of Gene Expression : Drivers of Natural Selection,"

正义-反义转录本的全基因组分析，“非编码 RNA 和基因表达的表观遗传调控：自然选择的驱动因素”，

DOI：
发表时间：
2012
期刊：
影响因子：
0
作者：
Kohama C;Kiyosawa H
通讯作者：
Kiyosawa H

Genome-wide analysis of expression modes and DNA methylation status at sense-antisense transcript loci in mouse.

DOI：
10.1016/j.ygeno.2010.08.007
发表时间：
2010-12
期刊：
Genomics
影响因子：
4.4
作者：
Yutaka Watanabe;K. Numata;S. Murata;Y. Osada;R. Saito;H. Nakaoka;Naoyuki Yamamoto;Kazufumi Watanabe;Hidemasa Kato;K. Abe;H. Kiyosawa
通讯作者：
Yutaka Watanabe;K. Numata;S. Murata;Y. Osada;R. Saito;H. Nakaoka;Naoyuki Yamamoto;Kazufumi Watanabe;Hidemasa Kato;K. Abe;H. Kiyosawa

シトルリン摂取による廃用性筋萎縮の抑制効果の検討.

检查瓜氨酸摄入对废用性肌肉萎缩的抑制作用。