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Palmitate-Induced Apoptosis of Microvascular Endothelial Cells and Pericytes

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Abstract

Background

Recent observations in the EURODIAB Complications Study demonstrated that markers of insulin resistance are strong risk factors for retinopathy incidence in patients with diabetes. However, the molecular mechanism underlying this remains to be elucidated. In this study, we investigated the influence of palmitate, a major saturated free fatty acid in plasma, on the apoptotic cell death of cultured microvascular endothelial cells (EC) and retinal pericytes.

Materials and Methods

The intracellular formation of reactive oxygen species (ROS) was detected using the fluorescent probe CM-H2DCFDA. DNA synthesis was determined by measuring [3H]-thymidine incorporation into cells. DNA fragmentations of EC were quantitatively analyzed in an enzyme-linked immunosorbent assay, and DNA laddering was evaluated on agarose gel electrophoresis.

Results

Palmitate increased ROS generation in microvascular EC. Furthermore, palmitate significantly inhibited DNA synthesis and induced apoptotic cell death in EC, which were completely prevented by an antioxidant, N-acetylcysteine. Palmitate up-regulated pericyte mRNA levels of a receptor for advanced glycation end products (AGE), and thereby potentiated the apoptotic effects of AGE on pericytes.

Conclusions

The results suggest that palmitate could induce apoptotic cell death in microvascular EC and pericytes through the overgeneration of intracellular ROS, and thus be involved in the development of diabetic retinopathy.

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Acknowledgments

This work was supported in part by Grants of Venture Research and Development Centers from the Ministry of Education, Culture, Sports, Science and Technology, Japan; the Suzuken Memorial Foundation, Japan; the Hukuriku University Foundation, Japan; and the Mochida Memorial Foundation for Medical and Pharmaceutical Research, Japan.

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Authors and Affiliations

  1. Division of Endocrinology and Metabolism, Department of Medicine, Kurume University School of Medicine, Kurume, 830-0011, Japan

    Sho-ichi Yamagishi, Tamami Okamoto, Shinjiro Amano, Yosuke Inagaki, Kohachiro Koga, Mari Koga & Zenji Makita

  2. Department of Neuropsychiatry, School of Medicine, Sapporo Medical University, Sapporo, Japan

    Hiroshi Choei & Nobuyuki Sasaki

  3. Department of Neurology, Hokkaido University Graduate School of Medicine, Sapporo, Japan

    Seiji Kikuchi

  4. Department of Biochemistry, Faculty of Pharmaceutical Science, Hokuriku University, Kanazawa, Japan

    Masayoshi Takeuchi

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  1. Sho-ichi Yamagishi
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  2. Tamami Okamoto
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  3. Shinjiro Amano
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  4. Yosuke Inagaki
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  5. Kohachiro Koga
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  6. Mari Koga
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Correspondence to Sho-ichi Yamagishi.

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Yamagishi, Si., Okamoto, T., Amano, S. et al. Palmitate-Induced Apoptosis of Microvascular Endothelial Cells and Pericytes. Mol Med 8, 179–184 (2002). https://doi.org/10.1007/BF03402010

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  • DOI: https://doi.org/10.1007/BF03402010

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