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High-glucose-increased expression and activation of NADPH oxidase in human vascular smooth muscle cells is mediated by 4-hydroxynonenal-activated PPARα and PPARβ/δ

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Abstract

High glucose induces vascular smooth muscle cell (SMC) dysfunction by generating oxidative stress attributable, in part, to the up-regulated NADPH oxidases (Nox). We have attempted to elucidate the high-glucose-generated molecular signals that mediate this effect and hypothesize that products of high-glucose-induced lipid peroxidation regulate Nox by activating peroxisome proliferator-activated receptors (PPARs). Human aortic SMCs were exposed to glucose (5.5–25 mM) or 4-hydroxynonenal (1–25 μM, 4-HNE). Lucigenin assay, real-time polymerase chain reaction, western blot, and promoter analyses were employed to investigate Nox. We found that high glucose generated an increase in Nox activity and expression. It also promoted oxidative stress that consequently induced lipid peroxidation, which resulted in the production of 4-HNE. Pharmacological inhibition of Nox activity significantly reduced the formation of high-glucose-induced 4-HNE. Exposure of SMCs to non-cytotoxic concentrations (1–10 μM) of 4-HNE alone mimicked the effect of high glucose incubation, whereas scavenging of 4-HNE by N-acetyl L-cysteine completely abolished both the effects of high glucose and 4-HNE. The latter exerted its effect by activating PPARα and PPARβ/δ, but not PPARγ, as assessed pharmacologically by the inhibitory effect of selective antagonists and following the silencing of the expression of these receptors. These new data indicate that 4-HNE, generated following Nox activation, functions as an endogenous activator of PPARα and PPARβ/δ. The newly discovered “lipid peroxidation products–PPARs–Nox axis” represents a novel mechanism of Nox regulation and an additional therapeutic target for oxidative stress in diabetes.

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Acknowledgments

We acknowledge the skillful assistance of Floarea Georgescu, Camelia Matei, and Constanta Stan.

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Authors and Affiliations

  1. Institute of Cellular Biology and Pathology “Nicolae Simionescu” of the Romanian Academy, 8 B.P. Hasdeu Street, 050568, Bucharest, Romania

    Adrian Manea, Simona-Adriana Manea, Andra Todirita, Irina Cristina Albulescu, Monica Raicu & Maya Simionescu

  2. “Petru Poni” Institute of Macromolecular Chemistry of the Romanian Academy, Iasi, Romania

    Adrian Manea

  3. The Institute for Drug Research, Department of Pharmacology, Faculty of Medicine, The Hebrew University, Jerusalem, Israel

    Shlomo Sasson

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  1. Adrian Manea
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  2. Simona-Adriana Manea
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  4. Irina Cristina Albulescu
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  5. Monica Raicu
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  7. Maya Simionescu
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Correspondence to Adrian Manea.

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This work was supported by the European Foundation for the Study of Diabetes – The New Horizons Grant to A. Manea in collaboration with S. Sasson, Romanian National Authority for Scientific Research (CNCS-UEFISCDI, project numbers PN-II-ID-PCE-2011-3-0548, PN-II-RU-TE-2011-3-0142, PNII-TE 65/2010), and COST action BM1203/EU-ROS. Simona A. Manea acknowledges the support of the strategic grant POSDRU/159/1.5/S/133391 financed by the European Social Found within the Sectorial Operational Program Human Resources Development 2007–2013. S. Sasson is the Adolf D. and Horty Storch Chair in Pharmaceutical Sciences, at the Faculty of Medicine, The Hebrew University of Jerusalem, Israel. He is affiliated with the David R. Bloom Center for Pharmacy and the Dr. Adolf and Klara Brettler Center for Research of Molecular Pharmacology and Therapeutics in the Hebrew University.

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Manea, A., Manea, SA., Todirita, A. et al. High-glucose-increased expression and activation of NADPH oxidase in human vascular smooth muscle cells is mediated by 4-hydroxynonenal-activated PPARα and PPARβ/δ. Cell Tissue Res 361, 593–604 (2015). https://doi.org/10.1007/s00441-015-2120-0

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