Transfection of naked oligodeoxynucleotides by rapid portal vein infusion

• Project/Area Number: 24591992
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Digestive surgery
• Research Institution: Akita University
• Principal Investigator: Go Watanabe 秋田大学, 医学(系)研究科(研究院), 助教 (50375276)
• Co-Investigator(Kenkyū-buntansha): YAMAMOTO Yuzo 秋田大学, 大学院医学系研究科, 教授 (70281730)
• Project Period (FY): 2012-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,070,000 (Direct Cost: ¥3,900,000、Indirect Cost: ¥1,170,000); Fiscal Year 2014: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000); Fiscal Year 2013: ¥1,170,000 (Direct Cost: ¥900,000、Indirect Cost: ¥270,000); Fiscal Year 2012: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
• Keywords: 遺伝子導入 / 肝虚血再灌流障害 / 虚血再灌流障害 / NF-κB / 遺伝子治療 / 肝臓外科

Outline of Final Research Achievements

This study was aimed to examine whether rapid portal vein infusion (RPVI) of naked oligodeoxynucleotides (ODN) could be used to transfect sufficient amounts of NF-kB decoy ODN into the liver to suppress NF-kB activation during liver ischemia/reperfusion (I/R) injury. Naked NF-kB decoy ODN solution was rapidly administered into the portal vein. Transfection efficacy was examined with a fluorescent tag. Activation of NF-kB was investigated by EMSA. Levels of serum liver enzymes and cytokines were measured during liver I/R injury. NF-kB decoy ODN was preferentially incorporated into Kupffer cells and sinusoidal endothelial cells, but not hepatocytes in the rat liver. Transfected NF-kB decoy ODN suppressed the function of NF-kB in both Kupffer cells and sinusoidal endothelial cells during liver I/R injury, causing significant decreases in serum TNF-a and IL-6 levels 3 h after reperfusion although the decrease of serum liver enzymes was not significant.