Rachel Hamias1, Assaf Rudich2,3, George Greenberg4, Gabriel Szendro4, Talya Wolak5. 1. Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev and Soroka University Medical Center, Beer Sheva, Israel. 2. Department of Clinical Biochemistry and Pharmacology, Faculty of Health Sciences, Ben-Gurion University of the Negev and Soroka Medical Center, Beer Sheva, Israel. 3. National Institute of Biotechnology, Negev, Israel. 4. Department of Vascular Surgery, Soroka University Medical Center, Beer-Sheva, Israel. 5. Hypertension Unit, Soroka University Medical Center and Ben-Gurion University of the Negev, P.O. Box 151, 84101, Beer-Sheva, Israel. twolak@bgu.ac.il.
Abstract
OBJECTIVE AND DESIGN: Evaluating the pro-/anti-inflammatory activity of the C-terminal cleavage product of osteopontin in comparison to angiotensin 1-7. MATERIAL AND SUBJECTS: Human coronary endothelial cells (hcEC) treated with conditioned media from human U937 macrophages. TREATMENT: Macrophages were (pre)treated with C-terminal, full-length or N-terminal osteopontin (OPN-C, OPN-FL, OPN-N, respectively), angiotensin II, angiotensin 1-7 or TNF-α. OPN-C modulatory capacity was compared to that of Ang1-7 in inhibiting subsequent Ag II, OPN-FL or OPN-N-induced macrophage-mediated endothelial inflammation. METHODS: Protein expression of NFκB, IκB, vCAM-1 and iCAM-1 was assessed using western blot. Promotor activation by NFκB was also assessed by dual-luciferase reporter assay. RESULTS: Conditioned media of macrophages treated with OPN-C induced hcECs' NfκB activation to a lower degree than OPN-FL or OPN-N. Priming of macrophages with angiotensin 1-7 attenuated the endothelial pro-inflammatory effect induced by subsequent exposure of the macrophages to angiotensin II, OPN-FL or OPN-N. This was evidenced by both NfκB activation and vCAM and iCAM expression. In contrast, priming macrophages with OPN-C did not significantly attenuate the subsequent response to the pro-inflammatory cytokines. CONCLUSIONS: OPN-C induces lower macrophage-induced endothelial inflammation compared to OPN-FL or OPN-N, but unlike angiotensin 1-7, fails to prevent endothelial inflammation induced by subsequent pro-inflammatory macrophage stimulation.
OBJECTIVE AND DESIGN: Evaluating the pro-/anti-inflammatory activity of the C-terminal cleavage product of osteopontin in comparison to angiotensin 1-7. MATERIAL AND SUBJECTS:Human coronary endothelial cells (hcEC) treated with conditioned media from human U937 macrophages. TREATMENT: Macrophages were (pre)treated with C-terminal, full-length or N-terminal osteopontin (OPN-C, OPN-FL, OPN-N, respectively), angiotensin II, angiotensin 1-7 or TNF-α. OPN-C modulatory capacity was compared to that of Ang1-7 in inhibiting subsequent Ag II, OPN-FL or OPN-N-induced macrophage-mediated endothelial inflammation. METHODS: Protein expression of NFκB, IκB, vCAM-1 and iCAM-1 was assessed using western blot. Promotor activation by NFκB was also assessed by dual-luciferase reporter assay. RESULTS: Conditioned media of macrophages treated with OPN-C induced hcECs' NfκB activation to a lower degree than OPN-FL or OPN-N. Priming of macrophages with angiotensin 1-7 attenuated the endothelial pro-inflammatory effect induced by subsequent exposure of the macrophages to angiotensin II, OPN-FL or OPN-N. This was evidenced by both NfκB activation and vCAM and iCAM expression. In contrast, priming macrophages with OPN-C did not significantly attenuate the subsequent response to the pro-inflammatory cytokines. CONCLUSIONS:OPN-C induces lower macrophage-induced endothelial inflammation compared to OPN-FL or OPN-N, but unlike angiotensin 1-7, fails to prevent endothelial inflammation induced by subsequent pro-inflammatory macrophage stimulation.
Authors: Jennifer Sullivan; Laurel Blair; Amer Alnajar; Tamer Aziz; Chee Yuan Ng; Galina Chipitsyna; Qiaoke Gong; Agnes Witkiewicz; Georg F Weber; David T Denhardt; Charles J Yeo; Hwyda A Arafat Journal: Surgery Date: 2009-08 Impact factor: 3.982