Bilirubin impairs bactericidal activity of neutrophils through an antioxidant mechanism in vitro

Toshiyuki Arai, Yasunobu Yoshikai, Junichi Kamiya, Masato Nagino, Katsuhiko Uesaka, Norihiro Yuasa, Koji Oda, Tsuyoshi Sano, Yuji Nimura

    Research output: Contribution to journalArticle

    26 Citations (Scopus)

    Abstract

    Background. Liver failure accompanied by hyperbilirubinemia after major hepatic resection is profoundly associated with septic complications. Although the immune dysfunction in cholestasis has been intensively investigated, the contribution of increased serum bilirubin to the impaired resistance to bacterial infection remains to be elucidated. Because bilirubin possesses an antioxidant activity, we hypothesized that bilirubin may scavenge reactive oxygen species (ROS) produced by neutrophils and consequently impair neutrophil bacterial killing. To address this, we evaluated the effects of bilirubin on the bactericidal activity of ROS or of neutrophils in vitro. Materials and methods. The antioxidant activity of bilirubin was determined using an ROS-sensitive fluorophore, dichlorofluorescin diacetate (DCFH-DA). Bilirubin concentration in the buffer solution was monitored spectorophotometrically after incubation with ROS. The effect of bilirubin on killing of Escherichia coli by ROS or by isolated human neutrophils was determined by counting the viable E. coli after incubation on nutrient agar. Results. The bilirubin concentration in the buffer solution was decreased by the addition of hydrogen peroxide, especially in the presence of peroxidase or ferrous iron. DCFH-DA oxidation by ROS or activated neutrophils was inhibited by bilirubin in a dose-dependent manner. The bactericidal activity of ROS or of isolated neutrophils was significantly attenuated by bilirubin. Conclusions. Bilirubin impairs bactericidal activity of neutrophils through scavenging ROS. Increased levels of serum bilirubin may well be responsible for the impaired bacterial clearance in patients with hyperbilirubinemia.

    Original languageEnglish
    Pages (from-to)107-113
    Number of pages7
    JournalJournal of Surgical Research
    Volume96
    Issue number1
    DOIs
    Publication statusPublished - Jan 1 2001

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    Bilirubin
    Neutrophils
    Antioxidants
    Reactive Oxygen Species
    Hyperbilirubinemia
    In Vitro Techniques
    Buffers
    Escherichia coli
    Cholestasis
    Liver Failure
    Serum
    Bacterial Infections
    Hydrogen Peroxide
    Peroxidase
    Agar
    Iron
    Food

    All Science Journal Classification (ASJC) codes

    • Surgery

    Cite this

    Arai, T., Yoshikai, Y., Kamiya, J., Nagino, M., Uesaka, K., Yuasa, N., ... Nimura, Y. (2001). Bilirubin impairs bactericidal activity of neutrophils through an antioxidant mechanism in vitro. Journal of Surgical Research, 96(1), 107-113. https://doi.org/10.1006/jsre.2000.6061

    Bilirubin impairs bactericidal activity of neutrophils through an antioxidant mechanism in vitro. / Arai, Toshiyuki; Yoshikai, Yasunobu; Kamiya, Junichi; Nagino, Masato; Uesaka, Katsuhiko; Yuasa, Norihiro; Oda, Koji; Sano, Tsuyoshi; Nimura, Yuji.

    In: Journal of Surgical Research, Vol. 96, No. 1, 01.01.2001, p. 107-113.

    Research output: Contribution to journalArticle

    Arai, T, Yoshikai, Y, Kamiya, J, Nagino, M, Uesaka, K, Yuasa, N, Oda, K, Sano, T & Nimura, Y 2001, 'Bilirubin impairs bactericidal activity of neutrophils through an antioxidant mechanism in vitro', Journal of Surgical Research, vol. 96, no. 1, pp. 107-113. https://doi.org/10.1006/jsre.2000.6061
    Arai, Toshiyuki ; Yoshikai, Yasunobu ; Kamiya, Junichi ; Nagino, Masato ; Uesaka, Katsuhiko ; Yuasa, Norihiro ; Oda, Koji ; Sano, Tsuyoshi ; Nimura, Yuji. / Bilirubin impairs bactericidal activity of neutrophils through an antioxidant mechanism in vitro. In: Journal of Surgical Research. 2001 ; Vol. 96, No. 1. pp. 107-113.
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    abstract = "Background. Liver failure accompanied by hyperbilirubinemia after major hepatic resection is profoundly associated with septic complications. Although the immune dysfunction in cholestasis has been intensively investigated, the contribution of increased serum bilirubin to the impaired resistance to bacterial infection remains to be elucidated. Because bilirubin possesses an antioxidant activity, we hypothesized that bilirubin may scavenge reactive oxygen species (ROS) produced by neutrophils and consequently impair neutrophil bacterial killing. To address this, we evaluated the effects of bilirubin on the bactericidal activity of ROS or of neutrophils in vitro. Materials and methods. The antioxidant activity of bilirubin was determined using an ROS-sensitive fluorophore, dichlorofluorescin diacetate (DCFH-DA). Bilirubin concentration in the buffer solution was monitored spectorophotometrically after incubation with ROS. The effect of bilirubin on killing of Escherichia coli by ROS or by isolated human neutrophils was determined by counting the viable E. coli after incubation on nutrient agar. Results. The bilirubin concentration in the buffer solution was decreased by the addition of hydrogen peroxide, especially in the presence of peroxidase or ferrous iron. DCFH-DA oxidation by ROS or activated neutrophils was inhibited by bilirubin in a dose-dependent manner. The bactericidal activity of ROS or of isolated neutrophils was significantly attenuated by bilirubin. Conclusions. Bilirubin impairs bactericidal activity of neutrophils through scavenging ROS. Increased levels of serum bilirubin may well be responsible for the impaired bacterial clearance in patients with hyperbilirubinemia.",
    author = "Toshiyuki Arai and Yasunobu Yoshikai and Junichi Kamiya and Masato Nagino and Katsuhiko Uesaka and Norihiro Yuasa and Koji Oda and Tsuyoshi Sano and Yuji Nimura",
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    AU - Arai, Toshiyuki

    AU - Yoshikai, Yasunobu

    AU - Kamiya, Junichi

    AU - Nagino, Masato

    AU - Uesaka, Katsuhiko

    AU - Yuasa, Norihiro

    AU - Oda, Koji

    AU - Sano, Tsuyoshi

    AU - Nimura, Yuji

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    N2 - Background. Liver failure accompanied by hyperbilirubinemia after major hepatic resection is profoundly associated with septic complications. Although the immune dysfunction in cholestasis has been intensively investigated, the contribution of increased serum bilirubin to the impaired resistance to bacterial infection remains to be elucidated. Because bilirubin possesses an antioxidant activity, we hypothesized that bilirubin may scavenge reactive oxygen species (ROS) produced by neutrophils and consequently impair neutrophil bacterial killing. To address this, we evaluated the effects of bilirubin on the bactericidal activity of ROS or of neutrophils in vitro. Materials and methods. The antioxidant activity of bilirubin was determined using an ROS-sensitive fluorophore, dichlorofluorescin diacetate (DCFH-DA). Bilirubin concentration in the buffer solution was monitored spectorophotometrically after incubation with ROS. The effect of bilirubin on killing of Escherichia coli by ROS or by isolated human neutrophils was determined by counting the viable E. coli after incubation on nutrient agar. Results. The bilirubin concentration in the buffer solution was decreased by the addition of hydrogen peroxide, especially in the presence of peroxidase or ferrous iron. DCFH-DA oxidation by ROS or activated neutrophils was inhibited by bilirubin in a dose-dependent manner. The bactericidal activity of ROS or of isolated neutrophils was significantly attenuated by bilirubin. Conclusions. Bilirubin impairs bactericidal activity of neutrophils through scavenging ROS. Increased levels of serum bilirubin may well be responsible for the impaired bacterial clearance in patients with hyperbilirubinemia.

    AB - Background. Liver failure accompanied by hyperbilirubinemia after major hepatic resection is profoundly associated with septic complications. Although the immune dysfunction in cholestasis has been intensively investigated, the contribution of increased serum bilirubin to the impaired resistance to bacterial infection remains to be elucidated. Because bilirubin possesses an antioxidant activity, we hypothesized that bilirubin may scavenge reactive oxygen species (ROS) produced by neutrophils and consequently impair neutrophil bacterial killing. To address this, we evaluated the effects of bilirubin on the bactericidal activity of ROS or of neutrophils in vitro. Materials and methods. The antioxidant activity of bilirubin was determined using an ROS-sensitive fluorophore, dichlorofluorescin diacetate (DCFH-DA). Bilirubin concentration in the buffer solution was monitored spectorophotometrically after incubation with ROS. The effect of bilirubin on killing of Escherichia coli by ROS or by isolated human neutrophils was determined by counting the viable E. coli after incubation on nutrient agar. Results. The bilirubin concentration in the buffer solution was decreased by the addition of hydrogen peroxide, especially in the presence of peroxidase or ferrous iron. DCFH-DA oxidation by ROS or activated neutrophils was inhibited by bilirubin in a dose-dependent manner. The bactericidal activity of ROS or of isolated neutrophils was significantly attenuated by bilirubin. Conclusions. Bilirubin impairs bactericidal activity of neutrophils through scavenging ROS. Increased levels of serum bilirubin may well be responsible for the impaired bacterial clearance in patients with hyperbilirubinemia.

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