Bradykinin- and lipopolysaccharide-induced bradykinin B₂ receptor expression, interleukin 8 release and ¡°nitrosative stress¡± in bronchial epithelial cells BEAS-2B: Role for neutrophils

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• 作者：Fabio L.M. Ricciardolo ; Valentina Sorbello ; Sabrina Benedetto ; Ilaria Defilippi ; Federica Sabatini ; Andrea Robotti ; Dein C. van Renswouw ; Caterina Bucca ; Gert Folkerts ; Virginia De Rose
• 关键词：Bradykinin ; Bradykinin B2 receptor ; Nitrosative stress ; Interleukin-8 ; Lipopolysaccharide ; Bronchial epithelium
• 刊名：European Journal of Pharmacology
• 出版年：2012
• 出版时间：5 November, 2012
• 年：2012
• 卷：694
• 期：1-3
• 页码：30-38
• 全文大小：766 K

文摘

Bradykinin-induced interleukin (IL)-8 release should potentially activate neutrophils releasing myeloperoxidase (MPO) and subsequently generating ¡°nitrosative stress¡±.

We studied bradykinin-induced expression of bradykinin B₂ receptor and bradykinin- and lipopolysaccharide (LPS)-induced IL-8 release, MPO (marker of neutrophil activation) and 3-nitrotyrosine (3-NT; marker of ¡°nitrosative stress¡±) production in human bronchial epithelial cells BEAS-2B alone or in co-culture with human neutrophils.

We evaluated B₂ receptor protein expression in BEAS-2B cells by immunostainings and Western blot analysis, and measured respectively bradykinin- or LPS-induced IL-8 release in BEAS-2B cells and bradykinin- and/or LPS-induced MPO and 3-NT production in BEAS-2B cells co-cultured with human neutrophils by ELISA. In addition, we evaluated bradykinin- and/or LPS-induced 3-NT formation in BEAS-2B cells co-cultured with human neutrophils by immunocytochemistry.

Bradykinin up-regulates B₂ receptor expression (P<0.05) and stimulate IL-8 release (P<0.001) in BEAS-2B cells. Either the selective bradykinin B₂ receptor antagonist HOE 140 or the selective bradykinin B₁ receptor antagonist Lys-(des-Arg⁹, Leu⁸)-bradykinin alone halved IL-8 release and the combination of both drugs suppressed this effect. In BEAS-2B cells co-cultured with human neutrophils bradykinin increased MPO release and 3-NT production compared to BEAS-2B cells with human neutrophils (P<0.001), and the addition of LPS in BEAS-2B cells with human neutrophils and bradykinin induced a further dramatically increase of MPO release and 3-NT formation (P<0.001).

Bradykinin and LPS provoked ¡°nitrosative stress¡±, potentially mediated by IL-8, in bronchial epithelium co-cultured with neutrophils suggesting a role for bradykinin in the amplification of chronic airway inflammation via production of ¡°nitrosative stress¡±.