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Anthocyanin-rich Seoritae extract ameliorates renal lipotoxicity via activation of AMP-activated protein kinase in diabetic mice.

Koh ES, Lim JH, Kim MY, Chung S, Shin SJ, Choi BS, Kim HW, Hwang SY, Kim SW, Park CW, Chang YS - J Transl Med (2015)

Bottom Line: Anthocyanins reversed diabetes-induced increases in renal apoptosis and oxidative stress.In cultured human glomerular endothelial cells, anthocyanins prevented high glucose-induced oxidative stress and apoptosis through activation of AMPK in the same manner.The results revealed that anthocyanins ameliorated diabetic nephropathy in db/db mice via phosphorylation of AMPK, the major energy-sensing enzyme, and the consequent effects on its target molecules, which appeared to prevent lipotoxicity-related apoptosis and oxidative stress in the kidney.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, The Catholic University of Korea Yeouido St. Mary's Hospital, 10, 63-ro, Yeongdeungpo-gu, Seoul, 150-713, Republic of Korea. fiji79@catholic.ac.kr.

ABSTRACT

Background: Anthocyanins are major constituents of food colours and have been reported to possess anti-diabetic activities for potential medicinal use. The precise role of anthocyanins in diabetic nephropathy is poorly understood. We investigated whether anthocyanin-rich Seoritae extract (SE) can potentially prevent oxidative stress and lipotoxicity, which are the main causes of renal damage in diabetic nephropathy, via activation of AMP-activated protein kinase (AMPK) and the consequent effects on its target molecules.

Methods: Four groups of male C57BLKS/J db/m and db/db mice were used. Diabetic and non-diabetic mice were orally administered 10 mg/kg body weight SE daily for 12 weeks, starting at 8 weeks of age.

Results: db/db mice treated with anthocyanins showed decreased albuminuria. Anthocyanins ameliorated intra-renal lipid concentrations in db/db mice with improvement of glomerular matrix expansion and inflammation, which was related to increased phosphorylation of AMPK and activation of peroxisome proliferator-activated receptor (PPAR) α and PPARγ, and inhibited the activity of acetyl-CoA carboxylase and sterol regulatory element-binding protein 1. Anthocyanins reversed diabetes-induced increases in renal apoptosis and oxidative stress. In cultured human glomerular endothelial cells, anthocyanins prevented high glucose-induced oxidative stress and apoptosis through activation of AMPK in the same manner.

Conclusions: The results revealed that anthocyanins ameliorated diabetic nephropathy in db/db mice via phosphorylation of AMPK, the major energy-sensing enzyme, and the consequent effects on its target molecules, which appeared to prevent lipotoxicity-related apoptosis and oxidative stress in the kidney.

No MeSH data available.


Related in: MedlinePlus

Effect of AMPKα1, AMPKα2, and SIRT1 siRNAs on anthocyanin-stimulated AMPK signalling in HGECs. Protein lysates (10 μg) from cultured HGECs were separated by SDS-PAGE and analysed by western blotting. Cultured HGECs were transfected with 40 nmol/L control siRNA or 40 nmol/L AMPKα1, AMPKα2, or SIRT1 siRNAs using a transfection reagent (Lipofectamine 2000). At approximately 24 h after transfection, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, and PGC-1α signalling in low-glucose medium were analysed. a–d Representative western blots (a) and quantitative analyses of the results b–d. *p < 0.05 and **p < 0.01 compared with the other groups. Cultured HGECs were transfected with 50 nmol/L control siRNA or 50 nmol/L AMPKΑ1, AMPKΑ2, or SIRT1 siRNAs and stimulated with anthocyanin in high-glucose medium. At approximately 24 h after stimulation, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, PPARα, PGC-1α, and ERR-1α signalling in high-glucose medium were analysed. e–j Representative western blots of phospho-AMPK Thr172, total AMPK, PPARα, PGC-1α, ERR-1α, and β-actin levels (e) and quantitative analyses of the results (f–j). *p < 0.05 and **p < 0.01 compared with high-glucose medium. HG high glucose.
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Fig6: Effect of AMPKα1, AMPKα2, and SIRT1 siRNAs on anthocyanin-stimulated AMPK signalling in HGECs. Protein lysates (10 μg) from cultured HGECs were separated by SDS-PAGE and analysed by western blotting. Cultured HGECs were transfected with 40 nmol/L control siRNA or 40 nmol/L AMPKα1, AMPKα2, or SIRT1 siRNAs using a transfection reagent (Lipofectamine 2000). At approximately 24 h after transfection, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, and PGC-1α signalling in low-glucose medium were analysed. a–d Representative western blots (a) and quantitative analyses of the results b–d. *p < 0.05 and **p < 0.01 compared with the other groups. Cultured HGECs were transfected with 50 nmol/L control siRNA or 50 nmol/L AMPKΑ1, AMPKΑ2, or SIRT1 siRNAs and stimulated with anthocyanin in high-glucose medium. At approximately 24 h after stimulation, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, PPARα, PGC-1α, and ERR-1α signalling in high-glucose medium were analysed. e–j Representative western blots of phospho-AMPK Thr172, total AMPK, PPARα, PGC-1α, ERR-1α, and β-actin levels (e) and quantitative analyses of the results (f–j). *p < 0.05 and **p < 0.01 compared with high-glucose medium. HG high glucose.

Mentions: To show whether the anti-apoptotic effect of anthocyanins is AMPK-dependent in diabetes, we performed additional experiments using siRNAs for AMPKΑ1, AMPKΑ2, and SIRT1 in cultured HGECs (Figure 6a–d). Transfected siRNAs for AMPKΑ1 and AMPKΑ2 suppressed anthocyanin-induced AMPK and PPARα–PGC-1α–ERR-1α signalling compared with the siRNA control group. However, transfection with the siRNA for Sirt1 only suppressed SIRT1, and not the phospho-Thr172 AMPK/total AMPK ratio and PPARα–PGC-1α–ERR-1α signalling (Figure 6e–j).Figure 6


Anthocyanin-rich Seoritae extract ameliorates renal lipotoxicity via activation of AMP-activated protein kinase in diabetic mice.

Koh ES, Lim JH, Kim MY, Chung S, Shin SJ, Choi BS, Kim HW, Hwang SY, Kim SW, Park CW, Chang YS - J Transl Med (2015)

Effect of AMPKα1, AMPKα2, and SIRT1 siRNAs on anthocyanin-stimulated AMPK signalling in HGECs. Protein lysates (10 μg) from cultured HGECs were separated by SDS-PAGE and analysed by western blotting. Cultured HGECs were transfected with 40 nmol/L control siRNA or 40 nmol/L AMPKα1, AMPKα2, or SIRT1 siRNAs using a transfection reagent (Lipofectamine 2000). At approximately 24 h after transfection, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, and PGC-1α signalling in low-glucose medium were analysed. a–d Representative western blots (a) and quantitative analyses of the results b–d. *p < 0.05 and **p < 0.01 compared with the other groups. Cultured HGECs were transfected with 50 nmol/L control siRNA or 50 nmol/L AMPKΑ1, AMPKΑ2, or SIRT1 siRNAs and stimulated with anthocyanin in high-glucose medium. At approximately 24 h after stimulation, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, PPARα, PGC-1α, and ERR-1α signalling in high-glucose medium were analysed. e–j Representative western blots of phospho-AMPK Thr172, total AMPK, PPARα, PGC-1α, ERR-1α, and β-actin levels (e) and quantitative analyses of the results (f–j). *p < 0.05 and **p < 0.01 compared with high-glucose medium. HG high glucose.
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Fig6: Effect of AMPKα1, AMPKα2, and SIRT1 siRNAs on anthocyanin-stimulated AMPK signalling in HGECs. Protein lysates (10 μg) from cultured HGECs were separated by SDS-PAGE and analysed by western blotting. Cultured HGECs were transfected with 40 nmol/L control siRNA or 40 nmol/L AMPKα1, AMPKα2, or SIRT1 siRNAs using a transfection reagent (Lipofectamine 2000). At approximately 24 h after transfection, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, and PGC-1α signalling in low-glucose medium were analysed. a–d Representative western blots (a) and quantitative analyses of the results b–d. *p < 0.05 and **p < 0.01 compared with the other groups. Cultured HGECs were transfected with 50 nmol/L control siRNA or 50 nmol/L AMPKΑ1, AMPKΑ2, or SIRT1 siRNAs and stimulated with anthocyanin in high-glucose medium. At approximately 24 h after stimulation, the levels of phospho-AMPK Thr172, total AMPK, SIRT1, PPARα, PGC-1α, and ERR-1α signalling in high-glucose medium were analysed. e–j Representative western blots of phospho-AMPK Thr172, total AMPK, PPARα, PGC-1α, ERR-1α, and β-actin levels (e) and quantitative analyses of the results (f–j). *p < 0.05 and **p < 0.01 compared with high-glucose medium. HG high glucose.
Mentions: To show whether the anti-apoptotic effect of anthocyanins is AMPK-dependent in diabetes, we performed additional experiments using siRNAs for AMPKΑ1, AMPKΑ2, and SIRT1 in cultured HGECs (Figure 6a–d). Transfected siRNAs for AMPKΑ1 and AMPKΑ2 suppressed anthocyanin-induced AMPK and PPARα–PGC-1α–ERR-1α signalling compared with the siRNA control group. However, transfection with the siRNA for Sirt1 only suppressed SIRT1, and not the phospho-Thr172 AMPK/total AMPK ratio and PPARα–PGC-1α–ERR-1α signalling (Figure 6e–j).Figure 6

Bottom Line: Anthocyanins reversed diabetes-induced increases in renal apoptosis and oxidative stress.In cultured human glomerular endothelial cells, anthocyanins prevented high glucose-induced oxidative stress and apoptosis through activation of AMPK in the same manner.The results revealed that anthocyanins ameliorated diabetic nephropathy in db/db mice via phosphorylation of AMPK, the major energy-sensing enzyme, and the consequent effects on its target molecules, which appeared to prevent lipotoxicity-related apoptosis and oxidative stress in the kidney.

View Article: PubMed Central - PubMed

Affiliation: Division of Nephrology, The Catholic University of Korea Yeouido St. Mary's Hospital, 10, 63-ro, Yeongdeungpo-gu, Seoul, 150-713, Republic of Korea. fiji79@catholic.ac.kr.

ABSTRACT

Background: Anthocyanins are major constituents of food colours and have been reported to possess anti-diabetic activities for potential medicinal use. The precise role of anthocyanins in diabetic nephropathy is poorly understood. We investigated whether anthocyanin-rich Seoritae extract (SE) can potentially prevent oxidative stress and lipotoxicity, which are the main causes of renal damage in diabetic nephropathy, via activation of AMP-activated protein kinase (AMPK) and the consequent effects on its target molecules.

Methods: Four groups of male C57BLKS/J db/m and db/db mice were used. Diabetic and non-diabetic mice were orally administered 10 mg/kg body weight SE daily for 12 weeks, starting at 8 weeks of age.

Results: db/db mice treated with anthocyanins showed decreased albuminuria. Anthocyanins ameliorated intra-renal lipid concentrations in db/db mice with improvement of glomerular matrix expansion and inflammation, which was related to increased phosphorylation of AMPK and activation of peroxisome proliferator-activated receptor (PPAR) α and PPARγ, and inhibited the activity of acetyl-CoA carboxylase and sterol regulatory element-binding protein 1. Anthocyanins reversed diabetes-induced increases in renal apoptosis and oxidative stress. In cultured human glomerular endothelial cells, anthocyanins prevented high glucose-induced oxidative stress and apoptosis through activation of AMPK in the same manner.

Conclusions: The results revealed that anthocyanins ameliorated diabetic nephropathy in db/db mice via phosphorylation of AMPK, the major energy-sensing enzyme, and the consequent effects on its target molecules, which appeared to prevent lipotoxicity-related apoptosis and oxidative stress in the kidney.

No MeSH data available.


Related in: MedlinePlus