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Antiatherosclerotic and renoprotective effects of ebselen in the diabetic apolipoprotein E/GPx1-double knockout mouse.

Chew P, Yuen DY, Stefanovic N, Pete J, Coughlan MT, Jandeleit-Dahm KA, Thomas MC, Rosenfeldt F, Cooper ME, de Haan JB - Diabetes (2010)

Bottom Line: The protective effects of ebselen were associated with a reduction in oxidative stress (hydroperoxides in plasma, 8-isoprostane in urine, nitrotyrosine in the kidney, and 4-hydroxynonenal in the aorta) as well as a reduction in VEGF, CTGF, VCAM-1, MCP-1, and Nox2 after 10 weeks of diabetes in the dKO aorta.Ebselen also significantly reduced the expression of proteins implicated in fibrosis and inflammation in the kidney as well as reducing related key intracellular signaling pathways.Ebselen has an antiatherosclerotic and renoprotective effect in a model of accelerated diabetic complications in the setting of enhanced oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Oxidative Stress Laboratory, Diabetes Division, Baker IDI Heart and Diabetes Institute, Melbourne, Australia.

ABSTRACT

Objective: To investigate the effect of the GPx1-mimetic ebselen on diabetes-associated atherosclerosis and renal injury in a model of increased oxidative stress.

Research design and methods: The study was performed using diabetic apolipoprotein E/GPx1 (ApoE(-/-)GPx1(-/-))-double knockout (dKO) mice, a model combining hyperlipidemia and hyperglycemia with increased oxidative stress. Mice were randomized into two groups, one injected with streptozotocin, the other with vehicle, at 8 weeks of age. Groups were further randomized to receive either ebselen or no treatment for 20 weeks.

Results: Ebselen reduced diabetes-associated atherosclerosis in most aortic regions, with the exception of the aortic sinus, and protected dKO mice from renal structural and functional injury. The protective effects of ebselen were associated with a reduction in oxidative stress (hydroperoxides in plasma, 8-isoprostane in urine, nitrotyrosine in the kidney, and 4-hydroxynonenal in the aorta) as well as a reduction in VEGF, CTGF, VCAM-1, MCP-1, and Nox2 after 10 weeks of diabetes in the dKO aorta. Ebselen also significantly reduced the expression of proteins implicated in fibrosis and inflammation in the kidney as well as reducing related key intracellular signaling pathways.

Conclusions: Ebselen has an antiatherosclerotic and renoprotective effect in a model of accelerated diabetic complications in the setting of enhanced oxidative stress. Our data suggest that ebselen effectively repletes the lack of GPx1, and indicate that ebselen may be an effective therapeutic for the treatment of diabetes-related atherosclerosis and nephropathy. Furthermore, this study highlights the feasibility of addressing two diabetic complications with one treatment regimen through the unifying approach of targeted antioxidant therapy.

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Ebselen abrogates H2O2-mediated increases in (A and D) P-p38; (B and E) P-IKK, and (C and F) P-JNK protein in normal rat kidney cells. A representative gel with its internal α-tubulin control (Aii, Biv, and Cvi) is shown above the quantitation for each protein. Lane 1 = untreated cells; lane 2 = serum-starved (SS) cells for 4 h; lane 3 = DMSO-treated cells; lane 4 = DMSO + 1 mmol/l H2O2-treated cells; lane 5 = 1 mmol/l H2O2 treated cells; lane 6 = 0.03 μmol/l ebselen-treated cells; and lane 7 = 1 mmol/l H2O2+0.03 μmol/l ebselen-treated cells. Arrows point to the two isoforms of P-JNK. Phosphorylated protein was quantitated relative to total protein (total protein levels are shown in supplementary online Fig. 11) for each gene. *P < 0.05 and **P < 0.01 vs. control cells, DMSO, and serum-starved cells. #P < 0.05 Ebselen plus H2O2 versus H2O2-treated cells. Bars, mean ± SEM; n = 4 replicates/group.
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Figure 8: Ebselen abrogates H2O2-mediated increases in (A and D) P-p38; (B and E) P-IKK, and (C and F) P-JNK protein in normal rat kidney cells. A representative gel with its internal α-tubulin control (Aii, Biv, and Cvi) is shown above the quantitation for each protein. Lane 1 = untreated cells; lane 2 = serum-starved (SS) cells for 4 h; lane 3 = DMSO-treated cells; lane 4 = DMSO + 1 mmol/l H2O2-treated cells; lane 5 = 1 mmol/l H2O2 treated cells; lane 6 = 0.03 μmol/l ebselen-treated cells; and lane 7 = 1 mmol/l H2O2+0.03 μmol/l ebselen-treated cells. Arrows point to the two isoforms of P-JNK. Phosphorylated protein was quantitated relative to total protein (total protein levels are shown in supplementary online Fig. 11) for each gene. *P < 0.05 and **P < 0.01 vs. control cells, DMSO, and serum-starved cells. #P < 0.05 Ebselen plus H2O2 versus H2O2-treated cells. Bars, mean ± SEM; n = 4 replicates/group.

Mentions: Initial experiments established optimal treatment time and dosage with H2O2 (30 min, at 1 mmol/l; data not shown) for increased expression of P-IKK, P-JNK, and P-p38. Pretreatment with 0.03 μmol/l ebselen for 30 min, followed by treatment with 1 mmol/l H2O2 for 30 min, significantly reduced H2O2-mediated increases in phosphorylation of p38, IKK, and JNK (Fig. 8; P < 0.05).


Antiatherosclerotic and renoprotective effects of ebselen in the diabetic apolipoprotein E/GPx1-double knockout mouse.

Chew P, Yuen DY, Stefanovic N, Pete J, Coughlan MT, Jandeleit-Dahm KA, Thomas MC, Rosenfeldt F, Cooper ME, de Haan JB - Diabetes (2010)

Ebselen abrogates H2O2-mediated increases in (A and D) P-p38; (B and E) P-IKK, and (C and F) P-JNK protein in normal rat kidney cells. A representative gel with its internal α-tubulin control (Aii, Biv, and Cvi) is shown above the quantitation for each protein. Lane 1 = untreated cells; lane 2 = serum-starved (SS) cells for 4 h; lane 3 = DMSO-treated cells; lane 4 = DMSO + 1 mmol/l H2O2-treated cells; lane 5 = 1 mmol/l H2O2 treated cells; lane 6 = 0.03 μmol/l ebselen-treated cells; and lane 7 = 1 mmol/l H2O2+0.03 μmol/l ebselen-treated cells. Arrows point to the two isoforms of P-JNK. Phosphorylated protein was quantitated relative to total protein (total protein levels are shown in supplementary online Fig. 11) for each gene. *P < 0.05 and **P < 0.01 vs. control cells, DMSO, and serum-starved cells. #P < 0.05 Ebselen plus H2O2 versus H2O2-treated cells. Bars, mean ± SEM; n = 4 replicates/group.
© Copyright Policy - creative-commons
Related In: Results  -  Collection

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Figure 8: Ebselen abrogates H2O2-mediated increases in (A and D) P-p38; (B and E) P-IKK, and (C and F) P-JNK protein in normal rat kidney cells. A representative gel with its internal α-tubulin control (Aii, Biv, and Cvi) is shown above the quantitation for each protein. Lane 1 = untreated cells; lane 2 = serum-starved (SS) cells for 4 h; lane 3 = DMSO-treated cells; lane 4 = DMSO + 1 mmol/l H2O2-treated cells; lane 5 = 1 mmol/l H2O2 treated cells; lane 6 = 0.03 μmol/l ebselen-treated cells; and lane 7 = 1 mmol/l H2O2+0.03 μmol/l ebselen-treated cells. Arrows point to the two isoforms of P-JNK. Phosphorylated protein was quantitated relative to total protein (total protein levels are shown in supplementary online Fig. 11) for each gene. *P < 0.05 and **P < 0.01 vs. control cells, DMSO, and serum-starved cells. #P < 0.05 Ebselen plus H2O2 versus H2O2-treated cells. Bars, mean ± SEM; n = 4 replicates/group.
Mentions: Initial experiments established optimal treatment time and dosage with H2O2 (30 min, at 1 mmol/l; data not shown) for increased expression of P-IKK, P-JNK, and P-p38. Pretreatment with 0.03 μmol/l ebselen for 30 min, followed by treatment with 1 mmol/l H2O2 for 30 min, significantly reduced H2O2-mediated increases in phosphorylation of p38, IKK, and JNK (Fig. 8; P < 0.05).

Bottom Line: The protective effects of ebselen were associated with a reduction in oxidative stress (hydroperoxides in plasma, 8-isoprostane in urine, nitrotyrosine in the kidney, and 4-hydroxynonenal in the aorta) as well as a reduction in VEGF, CTGF, VCAM-1, MCP-1, and Nox2 after 10 weeks of diabetes in the dKO aorta.Ebselen also significantly reduced the expression of proteins implicated in fibrosis and inflammation in the kidney as well as reducing related key intracellular signaling pathways.Ebselen has an antiatherosclerotic and renoprotective effect in a model of accelerated diabetic complications in the setting of enhanced oxidative stress.

View Article: PubMed Central - PubMed

Affiliation: Oxidative Stress Laboratory, Diabetes Division, Baker IDI Heart and Diabetes Institute, Melbourne, Australia.

ABSTRACT

Objective: To investigate the effect of the GPx1-mimetic ebselen on diabetes-associated atherosclerosis and renal injury in a model of increased oxidative stress.

Research design and methods: The study was performed using diabetic apolipoprotein E/GPx1 (ApoE(-/-)GPx1(-/-))-double knockout (dKO) mice, a model combining hyperlipidemia and hyperglycemia with increased oxidative stress. Mice were randomized into two groups, one injected with streptozotocin, the other with vehicle, at 8 weeks of age. Groups were further randomized to receive either ebselen or no treatment for 20 weeks.

Results: Ebselen reduced diabetes-associated atherosclerosis in most aortic regions, with the exception of the aortic sinus, and protected dKO mice from renal structural and functional injury. The protective effects of ebselen were associated with a reduction in oxidative stress (hydroperoxides in plasma, 8-isoprostane in urine, nitrotyrosine in the kidney, and 4-hydroxynonenal in the aorta) as well as a reduction in VEGF, CTGF, VCAM-1, MCP-1, and Nox2 after 10 weeks of diabetes in the dKO aorta. Ebselen also significantly reduced the expression of proteins implicated in fibrosis and inflammation in the kidney as well as reducing related key intracellular signaling pathways.

Conclusions: Ebselen has an antiatherosclerotic and renoprotective effect in a model of accelerated diabetic complications in the setting of enhanced oxidative stress. Our data suggest that ebselen effectively repletes the lack of GPx1, and indicate that ebselen may be an effective therapeutic for the treatment of diabetes-related atherosclerosis and nephropathy. Furthermore, this study highlights the feasibility of addressing two diabetic complications with one treatment regimen through the unifying approach of targeted antioxidant therapy.

Show MeSH
Related in: MedlinePlus