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Mitochondrial DNA mutations and oxidative damage in skeletal muscle of patients with chronic uremia

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Journal of Biomedical Science

Abstract

Abundant evidence has been gathered to suggest that mitochondrial DNA (mtDNA) sustains many more mutations and greater oxidative damage than does nuclear DNA in human tissues. Uremic patients are subject to a state of enhanced oxidative stress due to excess production of oxidants and a defective antioxidant defense system. This study was conducted to investigate mtDNA mutations and oxidative damage in skeletal muscle of patients with chronic uremia. Results showed that large-scale deletions between nucleotide position (np) 7,900 and 16,300 of mtDNA occurred at a high frequency in muscle of uremic patients. Among them, the 4,977-bp deletion (mtDNA4977) was the most frequent and most abundant large-scale mtDNA deletion in uremic skeletal muscle. The proportion of mtDNA4977 was found to correlate positively with the level of 8-hydroxy 2′-deoxyguanosine (8-OHdG) in the total DNA of skeletal muscle (r=0.62, p<0.05). Using long-range PCR and DNA sequencing, we identified and characterized multiple deletions of mtDNA in skeletal muscle of 16 of 19 uremic patients examined. The 8,041-bp deletion, which occurred between np 8035 and 16,075, was flanked by a 5-bp direct repeat of 5′-CCCAT-3′. Some of the deletions were found in more than 1 patient. On the other hand, we found that the mean 8-OHdG/105 dG ratio in the total cellular DNA of muscle of uremic patients was significantly higher than that of the controls (182.7 ± 63.6 vs. 50.9 ± 21.5, p=0.05). In addition, the mean 8-OHdG/105 dG ratio in muscle mtDNA of uremic patients was significantly higher than that in nuclear DNA (344.0 ± 56.9 vs. 146.3 ± 95.8, p=0.001). Moreover, we found that the average content of lipid peroxides in mitochondrial membranes of skeletal muscle of uremic patients was significantly higher than that of age-matched healthy subjects (23.76 ± 6.06 vs. 7.67 ± 0.95 nmol/mg protein; p<0.05). The average content of protein carbonyls in the mitochondrial membranes prepared from uremic skeletal muscles was significantly higher than that in normal controls (24.90 ± 4.00 vs. 14.48 ± 1.13 nmol/mg protein; p<0.05). Taken together, these findings suggest that chronic uremia leads to mtDNA mutations together with enhanced oxidative damage to DNA, lipids, and proteins of mitochondria in skeletal muscle, which may contribute to the impairment of mitochondrial bioenergetic function and to skeletal myopathy commonly seen in uremic patients.

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Authors and Affiliations

  1. Department of Biochemistry and Center for Cellular and Molecular Biology, National Yang-Ming University, Taipei

    Paik-Seong Lim, Yi-Shing Ma, Cheng-Feng Lee, Tzu-Ling Chen &  Yau-Huei Wei

  2. Graduate Institute of Clinical Medicinal Science, Chang Gung University, Taoyuan

    Paik-Seong Lim

  3. Department of Nephrology, Kuang Tien General Hospital, Taichung, Taiwan, Republic of China

    Paik-Seong Lim, Yueh-Mei Cheng & Henry Chai

  4. Department of Biochemistry, School of Life Sciences, National Yang-Ming University, Taipei, Taiwan 112 (ROC)

    Yau-Huei Wei

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Lim, PS., Ma, YS., Cheng, YM. et al. Mitochondrial DNA mutations and oxidative damage in skeletal muscle of patients with chronic uremia. J Biomed Sci 9, 549–560 (2002). https://doi.org/10.1007/BF02254982

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