2026-09-21T16:23:12Zhttps://repository.ajou.ac.kr/oai/request

oai:repository.ajou.ac.kr:201003/75712024-10-11T02:17:41Zcom_201003_14369com_201003_14368col_201003_14371

Study on the Neuroprotective Effects of Ethyl pyurvate in MPTP-induced Mouse PD Models MPTP로 유도된 파킨슨병 동물 모델에서 에틸 피루브산의 도파민성 신경세포사멸 억제 효과에 대한 연구 허, 수희 진, 병관 백, 은주 대학원 의학과 200524484 허, 수희 Parkinson`s disease MPTP Ethyl pyruvate glia ROS Neurotrophic factors The present study examined whether ethyl pyruvate (EP) promotes the survival of nigrostriatal dopaminergic (DA) neurons in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of Parkinson’s disease. MPTP induced degeneration of nigrostriatal DA neurons and glial activation as visualized by tyrosine hydroxylase, macrophage antigen complex-1, and/or glial fibrillary acidic protein immunoreactivity. Western blotting and immunohistochemistry showed activation of microglial NADPH oxidase and astroglial myeloperoxidase (MPO), and subsequent reactive oxygen species (ROS)/reactive nitrogen species (RNS) production and oxidative DNA damage in the MPTP-treated substantia nigra. Additional immunohistochemical staining also showed that MPTP induced glial cell line-derived neurotrophic factor (GDNF) and ciliary neurotrophic factor (CNTF) in the substantia nigra (SN). Interestingly, treatment with EP prevented degeneration of nigrostriatal DA neurons, increased striatal dopamine levels, and improved motor function. This neuroprotection afforded by EP was associated with the suppression of astroglial MPO expression, NADPH oxidase- and/or inducible nitric oxide synthase-derived ROS/RNS production by activated microglia. In parallel, treatment of EP significantly increased GDNF and CNTF in the MPTP-treated SN. Also, EP was found to protect DA neurons from 1-methyl-4-phenyl-pyridinium (MPP+) neurotoxicity in co-cultures of mesencephalic neurons and glia, but not in neuron-enriched mesencephalic cultures devoid of glia. The present findings show that EP may inhibit oxidative stress and increase levels of neurotrophic factors in glia, suggesting that EP may have therapeutic value in the treatment of aspects of Parkinson’s disease related to glia activation. ABSTRACT i TABLE OF CONTENTS iii LIST OF FIGURES ⅳ ABBREVIATION vii I. INTRODUCTION 1 A. Parkinson's disease 2 1. Characteristics of MPTP-induced mouse PD model 5 2. Role of glia in PD 8 3. Oxidative stress and PD 10 4. Neurotrophic factors and PD 12 B. Ethyl pyruvate 13 C. Purpose of present study 14 II. MATERIALS AND METHODS 16 A. Materials 16 1. Animals and treatment∙16 B. methods 16 1. Immunohistochemistry 16 2. Detection of dying neurons (Fluorojade-C staining) 17 3. Double-immunofluorescence Staining 18 4. Stereological cell counting 19 5. Densitometric analyses 20 6. Rotarod test 20 7. HPLC analyses 21 8. GC-MASS analyses 21 9. Determination of O2- and O2--derived oxidants 22 10. Immunoprecipitation 23 11. Western Blotting 23 12. Neuron-Enriched Mesencephalic Cultures 24 13. Mesencephalic Microglia Cultures 25 14. Co-cultures of Mesencephalic Microglia and Neurons 25 15. Measurement of Dopamine Uptake 26 16. Assays for intracellular ATP and ROS levels 26 17. Statistical analysis 27 III. RESULTS 28 Part A. Ethyl pyruvate (EP) protects nigrostriatal dopaminergic neuron from MPTP neurotoxicity 28 1. MPTP induces nigrostriatal DA neurodegeneration 28 2. MPTP induces glial activation in SN and STR. 30 3. EP protects nigrostriatal DA neurons from MPTP-induced neurotoxicity in vivo. 33 4. EP increases striatal dopamine levels and improves motor behavior in MPTP mice. 39 5. EP does not prevent MPP+-induced neurotoxicity in neuron-enriched mesencephalic cultures. 41 Part B. EP decreased oxidative stress increased by MPTP neurotoxicity 45 1. EP inhibits microglial activation in the SN in vivo 45 2. EP attenuates MPTP-induced reactive oxygen species (ROS) production and oxidative damage via microglial NADPH oxidase 48 3. EP attenuates iNOS expression and nitrosative damage 53 4. EP protects mesencephalic DA neurons from microglia-derived neurotoxicity 56 5. EP inhibits astrocytes activation in the SN in vivo∙ 59 6. EP inhibits myeloperoxydase (MPO) expression in astrocytes of the MPTP-treated SN in vivo 61 7. The expression of a variety of neurotrophic factors in the SN in MPTP-induced PD model 64 8. Pyruvate (Py) protects DA neuronal death in MPTP neurotoxicity 71 IV. DISCUSSION 73 V. CONCLUSION 82 REFERENCES 83 국문요약 100 Doctor 2012-10-29T04:47:53Z 2012-10-29T04:47:53Z 2012 2012 Thesis Theses http://repository.ajou.ac.kr/handle/201003/7571 http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000012408 000000012408 en