Enhanced muscle regeneration in mdx mice, Duchenne muscular dystrophy animal model, proven by CD44 & MYH3 expression, on oral feeding of N-163 strain of Aureobasidium Pullulans produced B-Glucan

Abstract

Introduction: Duchenne muscular dystrophy (DMD) is a X-linked fatal myogenic disorder, in which lack of dystrophin leads to significant fibrosis and a progressive loss of muscle mass. Regeneration of the muscles in response to the degeneration is not adequate to contribute to restoration of muscle function. Having proven the anti-inflammatory and anti-fibrotic properties of the 1,3-1,6 B-glucan (Neu REFIX) produced by the N-163 strain of Aureobasidium Pullulans in clinical and pre-clinical studies, we have now examined its effects on muscle regeneration in a DMD animal model of mdx mice. Methods: Forty-five mice were divided into the three groups, Gr. 1 (n=15): normal mice, Gr.2 (n=15): mdx mice as vehicle, Gr.3 (n=15) mdx mice administered the N-163 beta-glucan for 45 days. Expression of CD44 and Myosin heavy chain (MYH3) were evaluated in the skeletal muscles. Results: In the Gr.3 (N-163 group), CD44+ve staining in immunohistochemistry was more intense, with an H-score of > 2.0 as opposed to 1.0 in Gr. 2 and 0.1 in Gr. 1. Immunofluorescence staining showed 20% higher MYH3-staining positive area in Gr.3 (N-163 group) than that in the Gr.2. Conclusion: Successful muscle regeneration indicated by (i) increased MYH3 expression, characteristic of early myosin and muscle formation, followed by (ii) differentiation evident by an increase in CD44, upon oral feeding of N-163 B-Glucan, have been proven in this study. The earlier reported anti-fibrosis and anti-inflammatory effects in pre-clinical and human clinical studies along with the present study findings, unravel the potentials of this safe B-Glucan food supplement as a disease modifying adjunct in the management of DMD and other muscular dystrophies.