High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells

Toyoshi Inoguchi, Ping Li, Fumio Umeda, Hai Yan Yu, Maiko Kakimoto, Minako Imamura, Tsuyoshi Aoki, Takashi Etoh, Toshihiko Hashimoto, Masaichi Naruse, Hiroaki Sano, Hideo Utsumi, Hajime Nawata

    Research output: Contribution to journalArticle

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    Abstract

    Recent studies have revealed that vascular cells can produce reactive oxygen species (ROS) through NAD(P)H oxidase, which may be involved in vascular injury. However, the pathological role of vascular NAD(P)H oxidase in diabetes or in the insulin-resistant state remains unknown. In this study, we examined the effect of high glucose level and free fatty acid (FFA) (palmitate) on ROS production in cultured aortic smooth muscle cells (SMCs) and endothelial cells (ECs) using electron spin resonance spectroscopy. Exposure of cultured SMCs or ECs to a high glucose level (400 mg/dl) for 72 h significantly increased the free radical production compared with low glucose level exposure (100 mg/dl). Treatment of the cells for 3 h with phorbol myristic acid (PMA), a protein kinase C (PKC) activator, also increased free radical production. This increase was restored to the control value by diphenylene iodonium, a NAD(P)H oxidase inhibitor, suggesting ROS production through PKC-dependent activation of NAD(P)H oxidase. The increase in free radical production by high glucose level exposure was completely restored by both diphenylene iodonium and GF109203X, a PKC-specific inhibitor. Exposure to palmitate (200 μmol/l) also increased free radical production, which was concomitant with increases in diacylglycerol level and PKC activity. Again, this increase was restored to the control value by both diphenylene iodonium and GF109203X. The present results suggest that both high glucose level and palmitate may stimulate ROS production through PKC-dependent activation of NAD(P)H oxidase in both vascular SMCs and ECs. This finding may be involved in the excessive acceleration of atherosclerosis in patients with diabetes and insulin resistance syndrome.

    Original languageEnglish
    Pages (from-to)1939-1945
    Number of pages7
    JournalDiabetes
    Volume49
    Issue number11
    DOIs
    Publication statusPublished - Jan 1 2000

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    NADPH Oxidase
    Nonesterified Fatty Acids
    Protein Kinase C
    Blood Vessels
    Cultured Cells
    Reactive Oxygen Species
    Free Radicals
    Palmitates
    Glucose
    Smooth Muscle Myocytes
    Endothelial Cells
    Diacylglycerol Kinase
    Vascular System Injuries
    Electron Spin Resonance Spectroscopy
    Vascular Smooth Muscle
    Insulin Resistance
    Atherosclerosis
    Insulin
    diphenyleneiodonium
    bisindolylmaleimide I

    All Science Journal Classification (ASJC) codes

    • Internal Medicine
    • Endocrinology, Diabetes and Metabolism

    Cite this

    High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells. / Inoguchi, Toyoshi; Li, Ping; Umeda, Fumio; Yu, Hai Yan; Kakimoto, Maiko; Imamura, Minako; Aoki, Tsuyoshi; Etoh, Takashi; Hashimoto, Toshihiko; Naruse, Masaichi; Sano, Hiroaki; Utsumi, Hideo; Nawata, Hajime.

    In: Diabetes, Vol. 49, No. 11, 01.01.2000, p. 1939-1945.

    Research output: Contribution to journalArticle

    Inoguchi, T, Li, P, Umeda, F, Yu, HY, Kakimoto, M, Imamura, M, Aoki, T, Etoh, T, Hashimoto, T, Naruse, M, Sano, H, Utsumi, H & Nawata, H 2000, 'High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells', Diabetes, vol. 49, no. 11, pp. 1939-1945. https://doi.org/10.2337/diabetes.49.11.1939
    Inoguchi, Toyoshi ; Li, Ping ; Umeda, Fumio ; Yu, Hai Yan ; Kakimoto, Maiko ; Imamura, Minako ; Aoki, Tsuyoshi ; Etoh, Takashi ; Hashimoto, Toshihiko ; Naruse, Masaichi ; Sano, Hiroaki ; Utsumi, Hideo ; Nawata, Hajime. / High glucose level and free fatty acid stimulate reactive oxygen species production through protein kinase C-dependent activation of NAD(P)H oxidase in cultured vascular cells. In: Diabetes. 2000 ; Vol. 49, No. 11. pp. 1939-1945.
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    abstract = "Recent studies have revealed that vascular cells can produce reactive oxygen species (ROS) through NAD(P)H oxidase, which may be involved in vascular injury. However, the pathological role of vascular NAD(P)H oxidase in diabetes or in the insulin-resistant state remains unknown. In this study, we examined the effect of high glucose level and free fatty acid (FFA) (palmitate) on ROS production in cultured aortic smooth muscle cells (SMCs) and endothelial cells (ECs) using electron spin resonance spectroscopy. Exposure of cultured SMCs or ECs to a high glucose level (400 mg/dl) for 72 h significantly increased the free radical production compared with low glucose level exposure (100 mg/dl). Treatment of the cells for 3 h with phorbol myristic acid (PMA), a protein kinase C (PKC) activator, also increased free radical production. This increase was restored to the control value by diphenylene iodonium, a NAD(P)H oxidase inhibitor, suggesting ROS production through PKC-dependent activation of NAD(P)H oxidase. The increase in free radical production by high glucose level exposure was completely restored by both diphenylene iodonium and GF109203X, a PKC-specific inhibitor. Exposure to palmitate (200 μmol/l) also increased free radical production, which was concomitant with increases in diacylglycerol level and PKC activity. Again, this increase was restored to the control value by both diphenylene iodonium and GF109203X. The present results suggest that both high glucose level and palmitate may stimulate ROS production through PKC-dependent activation of NAD(P)H oxidase in both vascular SMCs and ECs. This finding may be involved in the excessive acceleration of atherosclerosis in patients with diabetes and insulin resistance syndrome.",
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    AU - Yu, Hai Yan

    AU - Kakimoto, Maiko

    AU - Imamura, Minako

    AU - Aoki, Tsuyoshi

    AU - Etoh, Takashi

    AU - Hashimoto, Toshihiko

    AU - Naruse, Masaichi

    AU - Sano, Hiroaki

    AU - Utsumi, Hideo

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