AIM: Senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice are incapable of synthesizing L-ascorbic acid (AA) in vivo. As AA is known to be a water-soluble anti-oxidant, we assessed protein oxidation levels in livers from SMP30/GNL KO mice maintained in an AA-insufficient condition. METHODS: Livers were collected from male SMP30/GNL KO mice at the ages of 3, 6 and 12 months, and wild-type (WT) mice at the ages of 3, 6, 12 and 24 months. To assess protein oxidation, we measured the content of protein carbonyl, which is a major protein oxidation marker. AA levels were measured by 2,4-dinitrophenylhydrazine method using high-performance liquid chromatography. RESULTS: Livers of SMP30/GNL KO mice had just ∼5% as much AA as those of WT mice from 3 to 12 months-of-age. Protein carbonyl levels in livers from SMP30/GNL KO mice were a significant 1.8- to 2.3-fold higher than those from age-atched WT mice. To establish that the AA-insufficiency caused this difference, we added AA to some drinking water, and examined the effect on AA and protein carbonyl levels in livers from SMP30/GNL KO and WT mice. Livers from SMP30/GNL KO mice given extra AA had a significantly higher content than those from their deprived counterparts. Furthermore, protein carbonyl levels in livers from AA-supplemented SMP30/GNL KO mice were significantly lower than those from the SMP30/GNL KO mice without AA supplementation. However, added AA did not affect the protein carbonyl levels in WT mice. CONCLUSIONS: These results strongly suggest that AA plays an important role in preventing protein oxidation in vivo, thus enhancing overall health.
AIM: Senescence marker protein-30 (SMP30)/gluconolactonase (GNL) knockout (KO) mice are incapable of synthesizing L-ascorbic acid (AA) in vivo. As AA is known to be a water-soluble anti-oxidant, we assessed protein oxidation levels in livers from SMP30/GNL KO mice maintained in an AA-insufficient condition. METHODS: Livers were collected from male SMP30/GNL KO mice at the ages of 3, 6 and 12 months, and wild-type (WT) mice at the ages of 3, 6, 12 and 24 months. To assess protein oxidation, we measured the content of protein carbonyl, which is a major protein oxidation marker. AA levels were measured by 2,4-dinitrophenylhydrazine method using high-performance liquid chromatography. RESULTS: Livers of SMP30/GNL KO mice had just ∼5% as much AA as those of WT mice from 3 to 12 months-of-age. Protein carbonyl levels in livers from SMP30/GNL KO mice were a significant 1.8- to 2.3-fold higher than those from age-atched WT mice. To establish that the AA-insufficiency caused this difference, we added AA to some drinking water, and examined the effect on AA and protein carbonyl levels in livers from SMP30/GNL KO and WT mice. Livers from SMP30/GNL KO mice given extra AA had a significantly higher content than those from their deprived counterparts. Furthermore, protein carbonyl levels in livers from AA-supplemented SMP30/GNL KO mice were significantly lower than those from the SMP30/GNL KO mice without AA supplementation. However, added AA did not affect the protein carbonyl levels in WT mice. CONCLUSIONS: These results strongly suggest that AA plays an important role in preventing protein oxidation in vivo, thus enhancing overall health.