Clinical use of functional regulation of aquaporin for brain edema decrease using knockdown technique

2017 Fiscal Year Final Research Report

• Project/Area Number: 15K10526
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Anesthesiology
• Research Institution: Aichi Medical University
• Principal Investigator: Fujita Yoshihito 愛知医科大学, 医学部, 教授 (90238593)
• Co-Investigator(Kenkyū-buntansha): 浅井 清文 名古屋市立大学, 大学院医学研究科, 教授 (70212462)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Keywords: 脳低温 / アストロサイt / アクアポリン / 水チャネル

Outline of Final Research Achievements

For making aquaporin (AQP) 4 knock down, we used Lentiviral RNAi system of Invitrogen corporation. Finally, we obtained AQP 4 knockdown model with pLenti4/BLOCK-IT/Expression Construct and plasmid of pLP1、pLP2、pLP/VSVG、and pENTER-gus. We confirmed that AQP4 knockdown that was stronger than our earlier experiment. We constructed hAQP4-M23 and hAQP4-M1 vector and transfected into C6 cells. We confirmed the overexpression of hAQP4-M1 and hAQP4-M23 in C6 cells. Moreover, we constructed permanent cell line of hAQP4-M23-overexpressed-cells of C6.In hypoxic condition, we tried to decide apoptosis or necrosis with flow cytometry. For optimal oxygen concentration, we experimented cell culture of astrocyte and SH-SY5Y in hyperoxia. We confirmed cell damage of hyperoxia in this condition. We try to continue neuron protection of various condition and drugs using RNAi technique.

Free Research Field

麻酔科学