title: Characterization of the Kallikrein-Kinin System Post Chemical Neuronal Injury: An In Vitro Biochemical and Neuroproteomics Assessment.
abstract: Traumatic Brain Injury (TBI) is the result of a mechanical impact on the brain provoking mild, moderate or severe symptoms. It is acknowledged that TBI leads to apoptotic and necrotic cell death; however, the exact mechanism by which brain trauma leads to neural injury is not fully elucidated. Some studies have highlighted the pivotal role of the Kallikrein-Kinin System (KKS) in brain trauma but the results are still controversial and inconclusive. In this study, we investigated both the expression and the role of Bradykinin 1 and 2 receptors (B1R and B2R), in mediating neuronal injury under chemical neurotoxicity paradigm in PC12 cell lines. The neuronal cell line PC12 was treated with the apoptotic drug Staurosporine (STS) to induce cell death. Intracellular calcium release was evaluated by Fluo 4-AM staining and showed that inhibition of the B2R prevented calcium release following STS treatment. Differential analyses utilizing immunofluorescence, Western blot and Real-time Polymerase Chain Reaction revealed an upregulation of both bradykinin receptors occurring at 3h and 12h post-STS treatment, but with a higher induction of B2R compared to B1R. This implies that STS-mediated apoptosis in PC12 cells is mainly conducted through B2R and partly via B1R. Finally, a neuroproteomics approach was conducted to find relevant proteins associated to STS and KKS in PC12 cells. Neuroproteomics results confirmed the presence of an inflammatory response leading to cell death during apoptosis-mediated STS treatment; however, a "survival" capacity was shown following inhibition of B2R coupled with STS treatment. Our data suggest that B2R is a key player in the inflammatory pathway following STS-mediated apoptosis in PC12 cells and its inhibition may represent a potential therapeutic tool in TBI.
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Genetics, American University of Beirut, Beirut, Lebanon.
authors.forename: Amaly
authors.initials: A
authors.lastname: Nokkari
authors.affiliation: Faculty of Medicine, American University of Beirut Medical Center, Beirut, Lebanon.
authors.forename: Tarek H
authors.initials: TH
authors.lastname: Mouhieddine
authors.affiliation: Faculty of Medicine, Saint George University of London, Nicosia, Cyprus.
authors.forename: Muhieddine M
authors.initials: MM
authors.lastname: Itani
authors.affiliation: Faculty of Medicine, Department of Anatomy, Cell Biology and Physiology, American University of Beirut, Beirut, Lebanon.
authors.forename: Wassim
authors.initials: W
authors.lastname: Abou-Kheir
authors.affiliation: Faculty of Medicine, Department of Anatomy, Cell Biology and Physiology, American University of Beirut, Beirut, Lebanon.
authors.forename: Georges
authors.initials: G
authors.lastname: Daoud
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas, United States of America.
authors.forename: Rui
authors.initials: R
authors.lastname: Zhu
authors.forename: Yehia
authors.initials: Y
authors.lastname: Mechref
authors.affiliation: Department of Chemistry and Biochemistry, Texas Tech University, Lubbock, Texas, United States of America.
authors.forename: Yehia
authors.initials: Y
authors.lastname: Meshref
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Genetics, American University of Beirut, Beirut, Lebanon.
authors.forename: Jihane
authors.initials: J
authors.lastname: Soueid
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Genetics, American University of Beirut, Beirut, Lebanon.
authors.forename: Moustafa
authors.initials: M
authors.lastname: Al Hariri
authors.affiliation: University of Messina, Department of Neurosciences, Messina, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Mondello
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Genetics, American University of Beirut, Beirut, Lebanon.
authors.forename: Ayad A
authors.initials: AA
authors.lastname: Jaffa
authors.affiliation: Faculty of Medicine, Department of Biochemistry and Molecular Genetics, American University of Beirut, Beirut, Lebanon.
authors.forename: Firas
authors.initials: F
authors.lastname: Kobeissy
mesh_terms: D000818:Animals; D016923:Cell Death / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000633:toxicity*; D016234:Kallikrein-Kinin System / Q000187:drug effects*; D009474:Neurons / Q000187:drug effects* / Q000276:immunology / Q000378:metabolism / Q000473:pathology*; D016716:PC12 Cells; D040901:Proteomics; D051381:Rats; D043783:Receptor, Bradykinin B1 / Q000032:analysis / Q000235:genetics / Q000276:immunology; D043782:Receptor, Bradykinin B2 / Q000032:analysis / Q000235:genetics / Q000276:immunology; D015398:Signal Transduction / Q000187:drug effects; D019311:Staurosporine / Q000633:toxicity*; D015533:Transcriptional Activation
chemical_list: D004791:Enzyme Inhibitors; D043783:Receptor, Bradykinin B1; D043782:Receptor, Bradykinin B2; D019311:Staurosporine