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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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The H2O2-mediated increase in Nox2 is reduced by ebselen in the diabetic ApoE−/−GPx1−/− (dKO) kidney. A: Quantitative RT-PCR analysis of Nox2 in ApoE−/− and dKO kidneys after 20 weeks of diabetes; n = 8–10 kidneys/group. B: Representative photomicrographs of kidney glomeruli (i–iv) and tubules (v–viii); nondiabetic dKO (i and v); diabetic dKO (ii and vi); diabetic dKO+Eb (iii and vii); and negative control which consisted of species-matched nonimmune IgG in place of primary antibody (iv and viii). C and D: Quantitation of Nox2 protein within glomeruli and tubules, respectively. ***P < 0.001; **P < 0.01; *P < 0.05 diabetic dKO vs. diabetic ApoE−/− kidneys; and versus nondiabetic counterparts. ##P < 0.01 and ###P < 0.001 vs. diabetic dKO kidneys. Bars, mean ± SEM; n = 20 glomeruli/kidney and 4–6 kidneys/group and n = 6 tubular fields/kidney and at least 5 kidneys/group. a.u., arbitrary units; D, diabetic; Eb, ebselen; ND, nondiabetic. (A high-quality digital representation of this figure is available in the online issue.)
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Figure 6: The H2O2-mediated increase in Nox2 is reduced by ebselen in the diabetic ApoE−/−GPx1−/− (dKO) kidney. A: Quantitative RT-PCR analysis of Nox2 in ApoE−/− and dKO kidneys after 20 weeks of diabetes; n = 8–10 kidneys/group. B: Representative photomicrographs of kidney glomeruli (i–iv) and tubules (v–viii); nondiabetic dKO (i and v); diabetic dKO (ii and vi); diabetic dKO+Eb (iii and vii); and negative control which consisted of species-matched nonimmune IgG in place of primary antibody (iv and viii). C and D: Quantitation of Nox2 protein within glomeruli and tubules, respectively. ***P < 0.001; **P < 0.01; *P < 0.05 diabetic dKO vs. diabetic ApoE−/− kidneys; and versus nondiabetic counterparts. ##P < 0.01 and ###P < 0.001 vs. diabetic dKO kidneys. Bars, mean ± SEM; n = 20 glomeruli/kidney and 4–6 kidneys/group and n = 6 tubular fields/kidney and at least 5 kidneys/group. a.u., arbitrary units; D, diabetic; Eb, ebselen; ND, nondiabetic. (A high-quality digital representation of this figure is available in the online issue.)

Mentions: We examined the gene expression of three isoforms of Nox (Nox 1, 2, 4) in the cortex of ApoE−/− and dKO mice (Fig. 6A and supplementary online Fig. 5). Expression was significantly increased in dKO kidneys (P < 0.001 for Nox2 and Nox4; and P < 0.05 for Nox1) compared with nondiabetic controls. Furthermore, diabetic dKO mRNA expression of Nox2 and Nox4 was significantly increased above that of diabetic ApoE−/− kidneys (P < 0.001). Nox2 protein was significantly increased in both the glomeruli and tubules (Fig. 6B) of diabetic dKO kidneys compared with nondiabetic controls (P < 0.05 for glomeruli; Fig. 6C and P < 0.01 for tubules; Fig. 6D), and was significantly increased compared with diabetic ApoE−/− kidneys (P < 0.05 for glomeruli and P < 0.01 for tubules). Ebselen significantly reduced the diabetes-driven increases in Nox2 (P < 0.001) and Nox4 (P < 0.001) expression in the diabetic dKO kidney, as well as Nox1 expression in the diabetic ApoE−/− kidney (P < 0.001). Furthermore, ebselen decreased Nox2 protein in the glomeruli and tubules (P < 0.01) of diabetic dKO kidneys.


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)

The H2O2-mediated increase in Nox2 is reduced by ebselen in the diabetic ApoE−/−GPx1−/− (dKO) kidney. A: Quantitative RT-PCR analysis of Nox2 in ApoE−/− and dKO kidneys after 20 weeks of diabetes; n = 8–10 kidneys/group. B: Representative photomicrographs of kidney glomeruli (i–iv) and tubules (v–viii); nondiabetic dKO (i and v); diabetic dKO (ii and vi); diabetic dKO+Eb (iii and vii); and negative control which consisted of species-matched nonimmune IgG in place of primary antibody (iv and viii). C and D: Quantitation of Nox2 protein within glomeruli and tubules, respectively. ***P < 0.001; **P < 0.01; *P < 0.05 diabetic dKO vs. diabetic ApoE−/− kidneys; and versus nondiabetic counterparts. ##P < 0.01 and ###P < 0.001 vs. diabetic dKO kidneys. Bars, mean ± SEM; n = 20 glomeruli/kidney and 4–6 kidneys/group and n = 6 tubular fields/kidney and at least 5 kidneys/group. a.u., arbitrary units; D, diabetic; Eb, ebselen; ND, nondiabetic. (A high-quality digital representation of this figure is available in the online issue.)
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Related In: Results  -  Collection

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Figure 6: The H2O2-mediated increase in Nox2 is reduced by ebselen in the diabetic ApoE−/−GPx1−/− (dKO) kidney. A: Quantitative RT-PCR analysis of Nox2 in ApoE−/− and dKO kidneys after 20 weeks of diabetes; n = 8–10 kidneys/group. B: Representative photomicrographs of kidney glomeruli (i–iv) and tubules (v–viii); nondiabetic dKO (i and v); diabetic dKO (ii and vi); diabetic dKO+Eb (iii and vii); and negative control which consisted of species-matched nonimmune IgG in place of primary antibody (iv and viii). C and D: Quantitation of Nox2 protein within glomeruli and tubules, respectively. ***P < 0.001; **P < 0.01; *P < 0.05 diabetic dKO vs. diabetic ApoE−/− kidneys; and versus nondiabetic counterparts. ##P < 0.01 and ###P < 0.001 vs. diabetic dKO kidneys. Bars, mean ± SEM; n = 20 glomeruli/kidney and 4–6 kidneys/group and n = 6 tubular fields/kidney and at least 5 kidneys/group. a.u., arbitrary units; D, diabetic; Eb, ebselen; ND, nondiabetic. (A high-quality digital representation of this figure is available in the online issue.)
Mentions: We examined the gene expression of three isoforms of Nox (Nox 1, 2, 4) in the cortex of ApoE−/− and dKO mice (Fig. 6A and supplementary online Fig. 5). Expression was significantly increased in dKO kidneys (P < 0.001 for Nox2 and Nox4; and P < 0.05 for Nox1) compared with nondiabetic controls. Furthermore, diabetic dKO mRNA expression of Nox2 and Nox4 was significantly increased above that of diabetic ApoE−/− kidneys (P < 0.001). Nox2 protein was significantly increased in both the glomeruli and tubules (Fig. 6B) of diabetic dKO kidneys compared with nondiabetic controls (P < 0.05 for glomeruli; Fig. 6C and P < 0.01 for tubules; Fig. 6D), and was significantly increased compared with diabetic ApoE−/− kidneys (P < 0.05 for glomeruli and P < 0.01 for tubules). Ebselen significantly reduced the diabetes-driven increases in Nox2 (P < 0.001) and Nox4 (P < 0.001) expression in the diabetic dKO kidney, as well as Nox1 expression in the diabetic ApoE−/− kidney (P < 0.001). Furthermore, ebselen decreased Nox2 protein in the glomeruli and tubules (P < 0.01) of diabetic dKO kidneys.

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