Markers of oxidative stress and aging in Duchene muscular dystrophy patients and the possible ameliorating effect of He:Ne laser

Abstract

Replicative aging and oxidative stress are two plausible theories explaining the etiology of muscular dystrophy. The first theory indicates that replicative aging of myogenic cells (satellite cells), owing to enhanced myofiber turnover, is a plausible explanation of the progression of Duchenne muscular dystrophy (DMD). The oxidative stress theory indicates that failure of muscle regeneration to keep up with the ongoing apoptosis and necrosis following oxidative stress, that normally associates muscular exercise, leads to muscle atrophy in DMD. To test for these two theories, markers of replicative aging and oxidative stress were assessed in the blood of 30 DMD patients vs. 20 normal healthy age matching controls. Markers of replicative aging showed significantly lower telomerase activity, significantly increased expression of receptors for advanced glycation end products (RAGEs) mRNA and Bax mRNA (an apoptotic gene) in DMD compared to controls. There was a significant increase in markers of oxidative stress among DMD patients compared to controls, measured in terms of increased apoptotic percentage in circulating mononuclear cells, increased lipid peroxidation measured in terms of plasma malondialdehyde (MDA) and increased protein carbonyls. Levels of plasma nitric oxide (NO), which neutralizes oxygen radicals, and expression of inducible nitric oxide synthase (iNOS) mRNA in neutrophils was significantly lower among DMD compared to controls. Biostimulation of WBC by helium neon (He:Ne) laser irradiation induced a significant increase in the expression of iNOS mRNA and plasma NO levels, but still at a lower level compared to controls. He:Ne laser irradiation induced a marked decrease in markers of oxidative stress among DMD patients compared to their level before irradiation, measured in terms of decreased plasma protein carbonyls, decreased plasma MDA, and decreased apoptosis percentage.

Conclusion: This study points to that oxidative stress is the prime cause for muscle degeneration in DMD and points out to the possible ameliorative effect of He:Ne laser on this

Figure 3

Bax mRNA expression among DMD patients compared to controls. Lanes 1-3 controls and lanes 4-6 DMD.

Figure 4

RAGEs mRNA expression. among DMD patients compared to controls. Lanes 1-4 controls and lanes 5-9 DMD.

Figure 5

Viable cells in circulating lymphocytes of lymphocytes a control case.

Figure 6

Apoptotic and viable cells in DMA patients.

Figure 1

Markers of replicative ageing among DMD patients compared to controls.

Figure 2

Markers of oxidative stress among DMD patients compared to controls and compared to its level post laser exposure.

Figure 7

Expression of iNOS mRNA pre and post laser irradiation, Lanes 1 and 3 for control pre and post laser respectively. Lanes 2 and 4 for DMD pre and post laser respectively.

Figure 8

Levels of inducible nitric oxide synthase among DMD patients compared to controls and compared to their levels post laser exposure.