Effects of Propofol on Platelet Activating Factor-Induced Microglial Activation

Other Titles
혈소판활성인자에 의해 유도된 미세아교세포 활성에 대한 Propofol의 영향
Authors
박종수
Issue Date
2010-12
Awarded Date
2011
Abstract
The aim of the present study was to investigate the effects of propofol on platelet activating factor (PAF)-induced microglial activation including intracellular Ca2+ level ([Ca2+]i), cell viability, reactive oxygen species (ROS) generation and canonical transient receptor potential Ca2+ channel (TRPC) expressions, and to evaluate the inhibitory mechanisms involved. PAF was used for activation of HMO6 human microglial cells. The changes in [Ca2+]i and ROS generation were measured by using fura-2/AM and 2',7'-dichlorofluorescin diacetate, respectively. 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyl tetrazolium bromide assay was used to evaluate cell viability. PAF-induced TRPC channel mRNA expression was analyzed by reverse transcription-polymerase chain reaction. PAF (300 nmol/L) induced an initial peak and sustained [Ca2+]i increases in human microglia. This [Ca2+]i increase was due to Ca2+ release from intracellular stores and Ca2+ influx through store-operated calcium (SOC) channel. PAF increased ROS generation whereas decreased cell viability. TRPC1 and TRPC4 mRNA expressions were significantly (p<0.05) increased after treatment of PAF. Propofol (10 μmol/L) inhibited SOC-mediated Ca2+ influx and ROS generation induced by PAF. Propofol recovered PAF-induced cell death to the levels of rotenone (mitochondrial respiratory chain inhibitor), but diphenyleneiodonium (NAPPH oxidase inhibitor) did not influence on the cell viability. Propofol significantly (p<0.05) attenuated the PAF-induced TRPC1 mRNA expression as potent as SOC channel blocker, 2-aminoethoxy diphenyl borate. These results suggest that inhibitory action mechanisms of propofol on PAF-induced microglial activation include inhibitions of SOC-mediated Ca2+ influx and mitochondrial ROS generation via a down-regulation of TRPC1 mRNA expression. 이 연구는 신경보호 작용 연구의 일환으로 혈소판활성인자(platelet activating factor, PAF)에 의해 미세아교세포가 활성화 되는 과정에서 propofol이 미치는 활성억제 효과를 검증하고 그 작용 기전을 파악하고자 하였다. 실험에 사용한 세포는 인체 유래 미세아교세포주인 HMO6세포를 이용하였다. 미세아교세포의 활성유도물질 중의 하나인 PAF (300 nmol/L)로 활성을 유도하였다. PAF에 의한 미세아교세포의 활성과 이에 대한 propofol (10 μmol/L)의 활성억제 효과를 fura-2 형광물질을 이용한 세포내 칼슘 농도 측정과 세포생존율, 활성산소 생성 및 역전사중합효소연쇄반응을 통한 canonical transient receptor potential Ca2+ channel (TRPC) 발현의 변화로 분석하였다. PAF는 미세아교세포의 세포내 칼슘저장고에서 칼슘유리를 유발시키고 store-operated calcium (SOC) 통로를 통한 세포 밖의 칼슘 유입을 촉진시켜 세포내 칼슘 농도를 증가시켰다. PAF는 활성산소의 생성을 증가시킨 반면에 세포생존율을 저하시켰다. PAF를 처리한 미세아교세포에서는 TRPC1 및 TRPC4 mRNA 발현이 유의하게(p<0.05) 증가되었다. Propofol은 PAF에 의해 유도된 SOC를 통한 칼슘 유입과 활성산소 생성을 억제하였다. 또한 propofol은 PAF에 의해 억제되었던 세포생존율을 회복시켰으며, TRPC1 mRNA를 유의하게(p<0.05) 억제하였다. 이상의 결과로 보아 propofol은 PAF에 의한 TRPC1 mRNA 발현 증가를 하향 조절함으로써 SOC를 통한 칼슘의 유입과 활성산소 생성을 차단하여 미세아교세포의 활성을 억제하는 것으로 생각된다.
URI
http://kumel.medlib.dsmc.or.kr/handle/2015.oak/11625
Appears in Collections:
3. Thesis (학위논문) > 1. School of Medicine (의과대학) > 박사
Full Text
http://dcollection.kmu.ac.kr//jsp/common/DcLoOrgPer.jsp?sItemId=000000009563
File in this Item
There are no files associated with this item.
Export
RIS (EndNote)
XLS (Excel)
XML


qrcode

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.

BROWSE