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SIRT1 reduction causes renal and retinal injury in diabetes through endothelin 1 and transforming growth factor β1.

Mortuza R, Feng B, Chakrabarti S - J. Cell. Mol. Med. (2015)

Bottom Line: Previously we showed glucose reduces sirtuin1 (SIRT1), a class III histone deacetylase.These cells also showed increased p300 expression, histone acetylation and reduced SIRT1 levels.These changes were rectified in the ECs following p300 silencing or by SIRT1 overexpression, whereas SIRT1 knockdown or p300 overexpression in NG mimicked the effects of HG.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Schulich School of Medicine & Dentistry, University of Western Ontario, London, ON, Canada.

No MeSH data available.


Related in: MedlinePlus

SIRT1 overexpression prevents glucose-induced increased endothelial permeability and collagen Iα(I) expression. (A) Duration dependent endothelial permeability and (B) end-point analysis showed, HG induced increased endothelial permeability was prevented by Ad-SIRT1 transfection or p300 siRNA treatment. SIRT1 siRNA or p300 overexpression in NG caused increased permeability by these cells mimicking the effects of HG. (C) HG caused an increase in Collagen Iα(I) mRNA expression in the ECs which was prevented by Ad-SIRT1 or p300 siRNA transfection. Opposingly, SIRT1 knockdown or p300 overexpression both lead to an up-regulation of Collagen Iα(I) mRNA levels in NG. (D) Western blot analysis of acetylated histone (Ac-H3K9/14) shows Ad-SIRT1 or p300 siRNA transfection reduced HG-induced increased histone acetylation in HMECs. On the other hand SIRT1 siRNA or p300 overexpression increased such acetylation in NG in these ECs. NG = 5 mmol/l glucose, HG = 25 mmol/l glucose, * = significantly different from NG, ╪ = significantly different from HG. HMECs  =  human microvascular endothelial cells. mRNA levels are expressed as a ratio of 18s. All data (mean ± SEM, P < 0.05) were normalized to NG; n = 4–6/group.
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fig03: SIRT1 overexpression prevents glucose-induced increased endothelial permeability and collagen Iα(I) expression. (A) Duration dependent endothelial permeability and (B) end-point analysis showed, HG induced increased endothelial permeability was prevented by Ad-SIRT1 transfection or p300 siRNA treatment. SIRT1 siRNA or p300 overexpression in NG caused increased permeability by these cells mimicking the effects of HG. (C) HG caused an increase in Collagen Iα(I) mRNA expression in the ECs which was prevented by Ad-SIRT1 or p300 siRNA transfection. Opposingly, SIRT1 knockdown or p300 overexpression both lead to an up-regulation of Collagen Iα(I) mRNA levels in NG. (D) Western blot analysis of acetylated histone (Ac-H3K9/14) shows Ad-SIRT1 or p300 siRNA transfection reduced HG-induced increased histone acetylation in HMECs. On the other hand SIRT1 siRNA or p300 overexpression increased such acetylation in NG in these ECs. NG = 5 mmol/l glucose, HG = 25 mmol/l glucose, * = significantly different from NG, ╪ = significantly different from HG. HMECs  =  human microvascular endothelial cells. mRNA levels are expressed as a ratio of 18s. All data (mean ± SEM, P < 0.05) were normalized to NG; n = 4–6/group.

Mentions: To investigate the functional consequences of SIRT1 mediated ET-1 and TGF-β1 regulation we conducted trans-endothelial permeability assay as increased endothelial permeability is a characteristic alteration in early diabetic microangiopathy. Data from the in vitro permeability test showed that HG significantly increases ECs permeability and Ad-SIRT1 transfection prevented such leakage in these cells (Fig.3A and B). p300 siRNA transfection in HG also had similar effect showing reduction in glucose-induced leakage whereas; SIRT1 knockdown or p300 overexpression in NG both significantly increased the ECs permeability mimicking the HG (Fig.3A and B). In addition, p300 overexpression in HG following treatment with Ad-SIRT1 reversed the beneficiary effects of SIRT1 causing increased EC permeability whereas p300 knockdown in the same scenario did not alter the beneficiary effects of SIRT1 (Fig. S1E).


SIRT1 reduction causes renal and retinal injury in diabetes through endothelin 1 and transforming growth factor β1.

Mortuza R, Feng B, Chakrabarti S - J. Cell. Mol. Med. (2015)

SIRT1 overexpression prevents glucose-induced increased endothelial permeability and collagen Iα(I) expression. (A) Duration dependent endothelial permeability and (B) end-point analysis showed, HG induced increased endothelial permeability was prevented by Ad-SIRT1 transfection or p300 siRNA treatment. SIRT1 siRNA or p300 overexpression in NG caused increased permeability by these cells mimicking the effects of HG. (C) HG caused an increase in Collagen Iα(I) mRNA expression in the ECs which was prevented by Ad-SIRT1 or p300 siRNA transfection. Opposingly, SIRT1 knockdown or p300 overexpression both lead to an up-regulation of Collagen Iα(I) mRNA levels in NG. (D) Western blot analysis of acetylated histone (Ac-H3K9/14) shows Ad-SIRT1 or p300 siRNA transfection reduced HG-induced increased histone acetylation in HMECs. On the other hand SIRT1 siRNA or p300 overexpression increased such acetylation in NG in these ECs. NG = 5 mmol/l glucose, HG = 25 mmol/l glucose, * = significantly different from NG, ╪ = significantly different from HG. HMECs  =  human microvascular endothelial cells. mRNA levels are expressed as a ratio of 18s. All data (mean ± SEM, P < 0.05) were normalized to NG; n = 4–6/group.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig03: SIRT1 overexpression prevents glucose-induced increased endothelial permeability and collagen Iα(I) expression. (A) Duration dependent endothelial permeability and (B) end-point analysis showed, HG induced increased endothelial permeability was prevented by Ad-SIRT1 transfection or p300 siRNA treatment. SIRT1 siRNA or p300 overexpression in NG caused increased permeability by these cells mimicking the effects of HG. (C) HG caused an increase in Collagen Iα(I) mRNA expression in the ECs which was prevented by Ad-SIRT1 or p300 siRNA transfection. Opposingly, SIRT1 knockdown or p300 overexpression both lead to an up-regulation of Collagen Iα(I) mRNA levels in NG. (D) Western blot analysis of acetylated histone (Ac-H3K9/14) shows Ad-SIRT1 or p300 siRNA transfection reduced HG-induced increased histone acetylation in HMECs. On the other hand SIRT1 siRNA or p300 overexpression increased such acetylation in NG in these ECs. NG = 5 mmol/l glucose, HG = 25 mmol/l glucose, * = significantly different from NG, ╪ = significantly different from HG. HMECs  =  human microvascular endothelial cells. mRNA levels are expressed as a ratio of 18s. All data (mean ± SEM, P < 0.05) were normalized to NG; n = 4–6/group.
Mentions: To investigate the functional consequences of SIRT1 mediated ET-1 and TGF-β1 regulation we conducted trans-endothelial permeability assay as increased endothelial permeability is a characteristic alteration in early diabetic microangiopathy. Data from the in vitro permeability test showed that HG significantly increases ECs permeability and Ad-SIRT1 transfection prevented such leakage in these cells (Fig.3A and B). p300 siRNA transfection in HG also had similar effect showing reduction in glucose-induced leakage whereas; SIRT1 knockdown or p300 overexpression in NG both significantly increased the ECs permeability mimicking the HG (Fig.3A and B). In addition, p300 overexpression in HG following treatment with Ad-SIRT1 reversed the beneficiary effects of SIRT1 causing increased EC permeability whereas p300 knockdown in the same scenario did not alter the beneficiary effects of SIRT1 (Fig. S1E).

Bottom Line: Previously we showed glucose reduces sirtuin1 (SIRT1), a class III histone deacetylase.These cells also showed increased p300 expression, histone acetylation and reduced SIRT1 levels.These changes were rectified in the ECs following p300 silencing or by SIRT1 overexpression, whereas SIRT1 knockdown or p300 overexpression in NG mimicked the effects of HG.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, Schulich School of Medicine & Dentistry, University of Western Ontario, London, ON, Canada.

No MeSH data available.


Related in: MedlinePlus