A new strategy for brain protection during circulatory arrest

2000 Fiscal Year Final Research Report Summary

• Project/Area Number: 11671318
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Thoracic surgery
• Research Institution: Osaka University
• Principal Investigator: SAWA Yoshiki Osaka Univ Graduate School of Med Lecturer, 医学系研究科, 講師 (00243220)
• Co-Investigator(Kenkyū-buntansha): NISHIMURA Motonobu Osaka Univ Graduate School of Med Assistant Professor, 医学系研究科, 助手 (90291442) ; FUKUSHIMA Norihide Osaka Univ Graduate School of Med Assistant Professor, 医学系研究科, 助手 (30263247) ; OHTAKE Shigeaki Osaka Univ Graduate School of Med Lecturer, 医学系研究科, 講師 (50243209) ; SAKAKIDA Satoru Osaka Univ Graduate School of Med Assistant Professor, 医学系研究科, 助手 (90311753)
• Project Period (FY): 1999 – 2000
• Keywords: circulatory arrest / brain protection / NF-κB / NF-κB decoy

Research Abstract

Objectives : Recent studies have reported that cis element decoy oligodeoxynucleotides (ODNs) against nuclear factor-kappa B (NF-κB) block the activation of genes that mediate ischemic injury. To improve brain protection during circulatory arrest in cardiac surgery, we evaluated the efficacy of NF-κB decoy ODNs in preventing neuronal damage after global brain ischemia.
Methods : Hemagglutinating virus of Japan (HVJ)-liposome complex with FITC-labeled NF-κB decoy ODNs was injected via the carotid artery during 20 minutes of global brain ischemia in rats, to evaluate the efficacy of transfecting the decoy ODNs. The mRNA levels of several factors related to ischemic-reperfusion injury in the hippocampus were estimated using a real-time polymerase chain reaction (PCR) method 1-hour after reperfusion. Neuronal damage was evaluated by TUNEL staining and by immunohistochemical study of microtubule-associated protein 2 (MAP2) in the hippocampus CA-1 region seven days after ischemia.
Results : Introduction of the NF-κB decoy ODNs into rat brain neurons via the carotid artery during global brain ischemia was markedly successful. The PCR study showed that the transfected NF-κB decoy ODNs effectively inhibited the expression of tumor necrosis factor-α (TNF-α), interleukin-1 β (IL-1 β), and intracellular adhesion molecule-1 (ICAM-1) mRNA 1 hour after global brain ischemia. TUNEL staining and MAP2 immunohistochemistry showed that the transfected NF-κB decoy ODNs significantly attenuated the neuronal damage seven days after global brain ischemia.
Conclusions : Therapeutic transfection of NF-κB decoy ODNs during brain ischemia may be useful for attenuating neuronal damage, suggesting a strategy for cerebral protection against global ischemia.