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Subcutaneous injection, from birth, of epigallocatechin-3-gallate, a component of green tea, limits the onset of muscular dystrophy in mdx mice: a quantitative histological, immunohistochemical and electrophysiological study

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Abstract

Dystrophic muscles suffer from enhanced oxidative stress. We have investigated whether administration of an antioxidant, epigallocatechin-3-gallate (EGCG), a component of green tea, reduces their oxidative stress and pathophysiology in mdx mice, a mild phenotype model of human Duchenne-type muscular dystrophy. EGCG (5 mg/kg body weight in saline) was injected subcutaneously 4× a week into the backs of C57 normal and dystrophin-deficient mdx mice for 8 weeks after birth. Saline was injected into normal and mdx controls. EGCG had almost no observable effects on normal mice or on the body weights of mdx mice. In contrast, it produced the following improvements in the blood chemistry, muscle histology, and electrophysiology of the treated mdx mice. First, the activities of serum creatine kinase were reduced to normal levels. Second, the numbers of fluorescent lipofuscin granules per unit volume of soleus and diaphragm muscles were significantly decreased by about 50% compared to the numbers in the corresponding saline-treated controls. Third, in sections of diaphragm and soleus muscles, the relative area occupied by histologically normal muscle fibres increased significantly 1.5- to 2-fold whereas the relative areas of connective tissue and necrotic muscle fibres were substantially reduced. Fourth, the times for the maximum tetanic force of soleus muscles to fall by a half increased to almost normal values. Fifth, the amount of utrophin in diaphragm muscles increased significantly by 17%, partially compensating for the lack of dystrophin expression.

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Acknowledgments

The authors thank Dr. Jun Ishibashi and Dr. Akio Hiura (University of Tokushima Graduate School, Japan), and Ms. Akiko Oka (Daiich Kikai Co., Ltd, Tokushima, Japan) for their instruction of Western blotting, mouse injection techniques and real-time RT-PCR, and Professor Juichiro Osame (Intellectual Property Office, University of Tokushima) for his continuous encouragement. We are also grateful for constructive comments from Professor Urs T. Ruegg (University of Geneva, Switzerland). This work was supported by research grants to YN from the Japan Society for the Promotion of Science and the Japan Science and Technology Agency.

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  1. Yoshiko Nakae

    Present address: Laboratory of Pharmacology, Geneva-Lausanne School of Pharmaceutical Sciences, University of Geneva, Sciences II 4-438B, 30 Quai Ernest-Ansermet, CH-1211, Geneva 4, Switzerland

Authors and Affiliations

  1. Department of Oral and Maxillofacial Anatomy, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, 770-8504, Japan

    Yoshiko Nakae

  2. Department of Nutritional Physiology, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, 770-8503, Japan

    Katsuya Hirasaka, Junpei Goto & Takeshi Nikawa

  3. Support Centre for Advanced Medical Sciences, Institute of Health Biosciences, The University of Tokushima Graduate School, Tokushima, 770-8503, Japan

    Masayuki Shono

  4. Department of Degenerative Neurological Diseases, National Institute of Neuroscience, National Centre of Neurology and Psychiatry, Kodaira, 187-8502, Japan

    Mizuko Yoshida

  5. College of Life Sciences, University of Dundee, Dundee, DD1 4HN, UK

    Peter J. Stoward

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  1. Yoshiko Nakae
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Correspondence to Yoshiko Nakae.

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Nakae, Y., Hirasaka, K., Goto, J. et al. Subcutaneous injection, from birth, of epigallocatechin-3-gallate, a component of green tea, limits the onset of muscular dystrophy in mdx mice: a quantitative histological, immunohistochemical and electrophysiological study. Histochem Cell Biol 129, 489–501 (2008). https://doi.org/10.1007/s00418-008-0390-2

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