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oai:repository.ajou.ac.kr:201003/13932024-10-11T02:17:35Zcom_201003_14369com_201003_14368col_201003_14370

이, 용범 이, 환구 대학원 의학과 200624465 이, 환구 2011-01-31T06:42:44Z 2011-01-31T06:42:44Z 2008 2008 http://repository.ajou.ac.kr/handle/201003/1393 http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000009208 000000009208 It is well known that excessive brain inflammation cause both acute and chronic neurodegeneration. Our previous study demonstrated the neuroprotective effects of IL-10 in LPS-injected rat cerebral cortex, which was mediated by inhibition of NADPH oxidase activation and pro-inflammatory mediators expression. Here, we observed the expression of pro-inflammatory mediators and IL-10 in the cerebral cortex of transgenic mouse model of AD (APP/PS1). In transgenic mice, microglia and astrocytes were activated synchronously with Aβ deposits and were abundant and closely associated with senile plaques. At 14-17 months, microglia and astrocytes were morphologically damaged and excessively activated. The mRNA and protein levels of IL-1β, TNF-α and iNOS were detected from 3 months, and they were increased with age. The mRNA and protein levels of IL-10 were detectable in 6 month-old transgenic mouse, which is 3 months later than the expression of pro-inflammatory mediators. Double immunoflorescence staining showed that IL-1β, TNF-α and IL-10 expression was localized mainly in Iba1-immunopositive microglia. Reactive oxygen species (ROS) production was detected from 6 months, and there most significantly increased between the ages of 14 months and 17 months. A major subunit of NADPH oxidase, gp91phox protein was localized to Iba1-immunopositive microglia in the APP/PS1 transgenic mice. And also, cerebral cortex of 14, 17 months showed significant decrease in NeuN-positive neurons and MAP2-immunoreactice dendrites. These results suggest that activated microglia surrounding plaques induce excessive production of pro-inflammatory mediators and ROS, leading to neuronal damage. 본 연구에서는 알츠하이머 동물모델 (APP/PS1 Transgenic Mice) 대뇌피질에서 염증성 매개물질인 IL-1β, TNF-α, iNOS, ROS와 IL-10의 발현을 관찰하였다. 유전자이식 생쥐에서 우리는 6개월부터 17개월에 이르기까지 섬유성 아밀로이드 베타 (fibrillar β-amyloid)의 축적이 늘어난다는 것을 확인하였다. 마이크로글리아와 아스트로사이트의 활성화는 아밀로이드 베타의 침착과 같은 양상으로 계속해서 증가되며, 노인성 반 (senile plaque)과 면밀하게 접근되어 나타나며, 14개월에서 17개월의 마이크로글리아와 아스트로사이트는 형태학적으로 손상되어 있으며 과도하게 활성화되어 있었다. IL-1β, TNF-α, iNOS의 mRNA와 단백질 레벨은 3개월부터 확인할 수 있으며 시간에 따라 증가된다. IL-10의 mRNA와 단백질 레벨은 6개월째 유전자이식 생쥐에서 확인할 수 있는데, 이는 염증성 매개물질들의 발현보다 3개월 늦은 것이다. 이중면역형광염색 (double immunoflorescence staining)으로 IL-1β, TNF-α, IL-10의 발현이 Iba1-면역양성을 보이는 마이크로글리아에서 대부분 배치됨을 관찰하였다. 활성산소종 (ROS) 생성은 8개월부터 나타나며 14개월과 17개월 사이에서 두드러지게 증가되었다. NADPH oxidase의 주요 구성물질인 gp91phox 단백질은 Iba1-면역양성을 보이는 마이크로글리아에서 대부분 배치됨을 관찰하였다. 또한, 14개월과 17개월의 대뇌피질에서 NeuN-양성적 뉴론과 MAP2-면역반응적인 수지상 돌기가 상당히 감소되어 있음을 관찰하였다. 이 결과들은 알츠하이머병 뇌조직에서 섬유성 아밀로이드 베타 (fibrillar β-amyloid)의 축적에 따라, 활성화된 마이크로글리아가 염증성 매개물질의 과도한 발현과 NADPH oxidase 활성화에 의한 활성산소종 생성을 유도하여 뉴론의 손상을 야기시킬 수 있음을 보여준다. Approved for entry into archive by Hyun Chul Cho (hccho@ajou.ac.kr) on 2011-01-31T06:42:44Z (GMT) No. of bitstreams: 1 000000009208.pdf: 1189041 bytes, checksum: bed506c4f3f29bf2eea1baea63771371 (MD5) Made available in DSpace on 2011-01-31T06:42:44Z (GMT). No. of bitstreams: 1 000000009208.pdf: 1189041 bytes, checksum: bed506c4f3f29bf2eea1baea63771371 (MD5) Submitted by Hyun Chul Cho (hccho@ajou.ac.kr) on 2011-01-31T06:42:44Z No. of bitstreams: 1 000000009208.pdf: 1189041 bytes, checksum: bed506c4f3f29bf2eea1baea63771371 (MD5) "ACKNOWLEDGEMENTS = ⅰ ABSTRACT = ⅱ TABLE OF CONTENTS = ⅳ LIST OF FIGURES = ⅵ LIST OF ABBREVIATION = ⅶ Ⅰ. INTRODUCTION = 1 A. Brain inflammation = 1 B. Brain inflammation in Alzheimer’s disease = 2 C. Aims of study = 3 Ⅱ. MATERIALS AND METHODS = 4 A. Transgenic mice and genotyping = 4 B. Immunohistochemistry = 4 C. Double-immunofluorescence staining = 5 D. Thioflavin S staining = 6 E. Reverse transcription-polymerase chain reaction (RT-PCR) = 6 F. In situ detection of O₂- and O₂--derived oxidants = 7 G. Western blot analysis = 8 H. Statistical analysis = 9 Ⅲ. RESULTS = 10 A. Age-related Aβ deposition and Aβ-associated gliosis in the cerebral cortex of APP/PS1 mice = 10 B. Expression of pro-inflammatory mediators and IL-10 in the cerebral cortex of APP/PS1 mice = 13 C. IL-1β, TNF-α, IL-10 expression in microglia = 15 D. ROS production and neuronal damage in the cerebral cortex of APP/PS1 mice = 19 Ⅳ. DISCUSSION = 22 Ⅴ. CONCLUSION = 25 REFERENCES = 26 국문요약 = 37" Master en Studies on the production of IL-1β, TNF-α, iNOS, IL-10, and ROS in the APP/PS1 mouse model of Alzheimer's disease 알츠하이머 동물모델 (APP/PS1 Transgenic Mice)에서 IL-1β, TNF-α, iNOS, IL-10, ROS 생성에 대한 연구 Thesis Theses Alzheimer’s disease Interleukin 10 microglia IL-1β TNF-α iNOS reactive oxygen species 2008 9999-12-31 9999-12-31