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병원성 파울러자유아메바 lysate에 의한 Primary rat astrocyte의 염증반응
- Alternative Title
- Inflammatory response of primary rat astrocyte treated with Naegleria fowleri lysate
- Author(s)
- 송아름
- Alternative Author(s)
- A Rum Song
- Advisor
- 신호준
- Department
- 일반대학원 분자과학기술학과
- Publisher
- The Graduate School, Ajou University
- Publication Year
- 2010-02
- Language
- eng
- Keyword
- Naegleria fowleri; astrocyte; inflammatory response; IL-1β; IL-6; MAPK; NF-κB; AP-1
- Alternative Abstract
- Naegleria fowleri, a free-living ameba, was found in diverse habitats throughout the world, and causes an acute, fulminating hemorrhagic meningoencephalitis, called primary amoebic meningoencephalitis (PAME), in healthy children and young adults with a history of recent exposure to polluted freshwater. Infection with N. fowleri is initiated by the introduction of water contaminating organisms into the nasal cavity of the host. Amoeba attachs to the nasal mucosa, migrate along the olfactory nerves, cross the blood-brain-barrier, and enter the brain. Astrocytes, abundant cells of CNS, involve the host’s defense against infection and then produce the inflammatory response. However, the inflammatory responses of astrocyte that is response to N. fowelri are largely unknown up to recently. In this study, to understand microbes from the infected CNS, as well as in the immunopathogenesis of inflammatory diseases, we focus both on how cytokines and immune response can be activated. Using RPA (RNase protection assay) and RT-PCR, we revealed that IL-1β and IL-6 mRNA expression were increased after N. fowleri lysate treatment. IL-6 mRNA expression is more strong than IL-1β. To identify relation between MAPK and expression of inflammatory cytokine, I performed RT-PCR using specific inhibitors. The expression levels of cytokine in response to stimuli were gradually decreased on account of ERK and P38, and JNK inhibitor (ERK inhibitor, U0126; P38 inhibitor, SB202190; JNK inhibitor, SP600125). And all MAPK inhibitors reduced the IL-1β and IL-6 protein levels in ELISA. But, activation of MAPK slightly detected by western blot in N. fowleri lysate-stimulated astrocytes. To find out transcription factor of these cytokine, I was used to transcription inhibitor (AP-1 inhibitor, Tanshinone IIA; NF-κB inhibitor, BAY 11-7082). Although mRNA expression of N. fowleri-induced IL-1β and IL-6 was elevated by MG132, their protein levels were significantly reduced by BAY 11-7082. In addition, experiments though Tanshinone IIA, were down-regulated IL-1β and IL-6 expression. From these results, I considered that both NF-κB and AP-1 related to production of pro-inflammatory cytokine, IL-1β and IL-6. Consequently when primary rat astrocyte treated with N. fowleri lysate, IL-1β, and IL-6 mRNA expression, and secretion elevated, and this process is adjusted through MAPKs, NF-κB, and AP-1. It may be implied that production of pro-inflammatory cytokine are likely to act an important role in the inflammatory response to N. fowleri infection in the CNS.
- Fulltext
- Appears in Collections:
- Graduate School of Ajou University > Department of Molecular Science and Technology > 3. Theses(Master)
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