2026-09-22T03:00:41Zhttps://repository.ajou.ac.kr/oai/request

oai:repository.ajou.ac.kr:201003/13822024-10-11T02:17:34Zcom_201003_14369com_201003_14368col_201003_14370

이, 필휴 김, 유정 대학원 의학과 200624165 김, 유정 2011-01-31T05:03:12Z 2011-01-31T05:03:12Z 2008 2008 http://repository.ajou.ac.kr/handle/201003/1382 http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000005879 000000005879 Parkinson's disease (PD) is a common progressive neurodegenerative disorder caused by the loss of dopaminergic neurons in the substantia nigra (SN). Numerous studies have provided evidence suggesting that neuroinflammation plays a critical role in the pathogenesis of PD. The present study was to evaluate effect of human bone marrow-derived mesenchymal stem cell (hMSCs) on modulation of neuroinflammation. We used lipopolysaccharide (LPS)-induced in vitro and in vivo inflammation models to investigate whether hMSCs have a protective effect on the dopaminergic system through anti-inflammatory mechanism. hMSCs treatment significantly decreased LPS-induced microglial activation and production of pro-inflammatory cytokines compared to LPS-only treatment group. In co-culture cells of microglia and mesencephalic dopaminergic neurons, hMSCs treatment resulted in a significant reduction of dopaminergic neuronal loss induced by LPS stimulation. In animal study, hMSCs treatment significantly increased survival of TH-ip cells at 7 and 14 days following hMSCS injection, approximately two times more than the LPS-only treatment, which was clearly accompanied by a decrease in activation of microglia. Additionally, cytokine assay in the SN showed that hMSCs treatment significantly down regulated the LPS-induced increase in the expression of TNF-α and iNOS mRNA following LPS stimulation. TNF-α and iNOS release were also significantly decreased in the hMSCs group when compared to the LPS-only treatment group at 4 hr and 3 day following hMSCS injection. The present study demonstrated that MSCs have a neuroprotective effect on dopaminergic neurons via an anti-inflammatory mechanism mediated by the modulation of microglial activation. Along with various trophic effect and transdifferentiational potency, anti-inflammatory mechanism of MSCs could have major therapeutic implication in the treatment of PD. 파킨슨 씨 병은 흑색질 안에 도파민 신경세포의 감소가 원인이 되는 퇴행성 뇌질환이다. 이미 여러 연구에서 신경염증이 파킨슨 씨 병의 발병에 중요한 역할을 함이 증명되었다. 현 우리 실험에서는 성체 중간엽 세포가 신경염증을 완화하는 효과를 확인하였다. 우리는 lipopolysaccharide (LPS)로 실헌 동물 모델과 세포 모델을 만들어 염증을 유도한 후 성체 중간엽 줄기세포가 항염증 메커니즘을 통하여 도파민 신경세포를 보호하는 효과를 갖는지 확인하였다. 성체 중간엽 줄기세포를 처리하였을 때 LPS만 처리한 개체군과 비교하여 LPS에 의해 유도된 전구염증 사이토카인의 증가와 미세아교세포의 활성이 유의미하게 감소함을 알 수 있었다. 또한 미세아교세포와 중뇌 도파민 신경세포의 co-culture system에서 성체 중간엽 줄기세포를 처리한 경우 LPS로 인한 도파민 신경세포의 감소가 현저히 줄어듦을 확인하였다. 그리고 동물 실험을 통하여 성체 중간엽 줄기세포를 주입하였을 때 성체 중간엽 줄기세포를 주입한지 7, 14일이 지난 후 TH-immunopositive (TH-ip)를 표현하는 세포가 증가함을 확인하였으며 이는 LPS 개체군에 비하여 그 양이 대략 두 배 이상이었고 흑색질에서 미세아교세포의 활성의 감소도 수반하였다. 그리고 흑색질 안의 사이토카인 정량 실험 결과, 성체 중간엽 줄기세포를 주입한 개체군은 LPS에 의해 증가된 TNF-α와 iNOS mRNA의 발현을 조절억제 되었고, 성체 중간엽 줄기세포가 주입된지 4시간과 3일 후의 개체군에서 TNF-α와 iNOS의 발현 역시 LPS만 투여된 개체군에 비교하여 유의미하게 감소됨을 관찰하였다. 본 연구는 성체 중간엽 줄기세포가 항염증 메커니즘을 통하여 미세아교세포의 활성을 조절함으로 도파민 신경세포를 보호하는 것을 확인하였고, 이는 다양한 trophic effect와 transdifferentiation 가능성과 함께 성체 중간엽 줄기세포의 항염증 메커니즘이 파킨슨 씨 병의 치료에 주요하게 사용될 수 있음을 시사한다. Approved for entry into archive by Hyun Chul Cho (hccho@ajou.ac.kr) on 2011-01-31T05:03:12Z (GMT) No. of bitstreams: 1 000000005879.pdf: 11096635 bytes, checksum: 971e987d30327a95ec8932ad4a77d739 (MD5) Made available in DSpace on 2011-01-31T05:03:12Z (GMT). No. of bitstreams: 1 000000005879.pdf: 11096635 bytes, checksum: 971e987d30327a95ec8932ad4a77d739 (MD5) Submitted by Hyun Chul Cho (hccho@ajou.ac.kr) on 2011-01-31T05:03:12Z No. of bitstreams: 1 000000005879.pdf: 11096635 bytes, checksum: 971e987d30327a95ec8932ad4a77d739 (MD5) "ABSTRACT = ⅰ TABLE OF CONTENTS = ⅱ LIST OF FIGURES = ⅳ Ⅰ. INTRODUCTION = 1 Ⅱ. MATERIALS AND METHODS = 3 A. MATERIALS = 3 B. METHODS = 4 1. Isolation of hMSCs and culture maintenance = 4 2. Co-culture of LPS treated enriched-microglia or neuron-glia culture with transwell hMSCs insert = 4 3. Microglia enriched cultures = 5 4. Co-cultures of microglia and mesencephalic neuron cultures = 5 5. Determination of NO release = 6 6. Animals study = 6 7. Reverse transcription PCR (RT-PCR) = 7 8. Quantification of TNF-α release = 8 9. Tissue preparation = 8 10. Immunocytochemistry and immunohistochemistry = 9 11. Stereological cell counts = 9 12. Statistical analysis = 10 Ⅲ. RESULTS = 11 1. hMSCs decreased LPS-induced microglial activation = 11 2. hMSCs treatment significantly decreased LPS-induced expression of inflammatory cytokine = 11 3. hMSCs treatment significantly reduced dopaminergic neuronal death induced by LPS stimulation in mesencephalic tissue and microglia co-cultured system = 12 4. hMSCs treatment significantly decreased dopaminergic neuronal loss and microglia activation induced by LPS stimulation in the SN = 12 5. hMSCs reduced the protein expression of TNF-α release induced by LPS stimulation = 13 Ⅳ. DISCUSSION = 23 REFERENCES = 27 국문요약 = 33" Master application/pdf en Neuroprotective Effect of Human Mesenchymal Stem Cell on Dopaminergic Neurons by Anti-inflammatory Action 성체 중간엽 줄기세포의 항염증 작용을 통한 도파민 신경세포의 보호 효과 Thesis Theses Human mesenchymal stem cell anti-inflammation dopaminergic neurons mMicroglia Parkinson’s disease 성체 중간엽 줄기세포 항염증 도파민 신경세포 미세아교세포 파킨슨씨병 2008 9999-12-31 9999-12-31