簡易檢索 / 詳目顯示

研究生: 吳嘉旋
Wu, Chia-Hsuan
論文名稱: 探討黃芩素衍生物於神經細胞及阿茲海默動物模式小鼠之作用
The effect of Baicalein derivatives on cortical neurons and on an Alzheimer’s disease mouse model
指導教授: 林炎壽
Lin, Yenshou
口試委員: 林炎壽
Lin, Yenshou
梁美智
Liang, Mei-Chih
李冠群
Lee, Guan-Chiun
口試日期: 2024/03/27
學位類別: 碩士
Master
系所名稱: 生命科學系
Department of Life Science
論文出版年: 2024
畢業學年度: 112
語文別: 中文
論文頁數: 67
中文關鍵詞: 阿茲海默症β類澱粉蛋白DiBAC4(3)Fura-2/AM
英文關鍵詞: Alzheimer's disease, amyloid beta, DiBAC4(3), Fura-2/AM
研究方法: 實驗設計法
DOI URL: http://doi.org/10.6345/NTNU202400444
論文種類: 學術論文
相關次數: 點閱:94下載:0
分享至:
查詢本校圖書館目錄 查詢臺灣博碩士論文知識加值系統 勘誤回報
  • 第一章 緒論 1 一、 阿茲海默症之β類澱粉蛋白形成 1 二、 大腦皮質之麩胺酸神經傳遞系統 2 三、 β類澱粉蛋白對麩胺酸傳遞影響 2 四、 阿茲海默症模式小鼠J20 3 五、 臨床治療阿茲海默症藥物 4 六、 中草藥黃芩之功效 5 第二章 研究目的 6 第三章 研究材料與實驗方法 7 一、 藥劑 7 二、 小鼠大腦皮質神經細胞初級培養 8 三、 細胞免疫染色 (Immunocytochemistry, ICC) 8 四、 β類澱粉蛋白寡聚體製備及蛋白質斑點印跡分析 (Dot blot assay) 9 五、 DiBAC4(3)膜電位染劑檢測去極化反應 10 六、 Fura-2/AM (Fura-2-acetoxy-methyl-ester) 鈣離子測量 11 七、 蛋白質萃取、西方墨點法 11 八、 基因型檢測 12 九、 實驗動物與藥物給予 13 十、 莫氏水迷津測試 (Morris water maze) 14 十一、 巴恩斯迷宮 (Barnes maze) 14 十二、 新物體辨識 (Novel Object Recognition) 15 十三、 臉頰採血檢測血清中的GOT、GPT 16 十四、 灌流 (Perfusion) 及肝臟、腎臟組織H&E染色 16 十五、 統計分析 17 第四章 研究結果 18 一、 BE3能抑制細胞因Aβ所誘發的神經細胞不正常去極化 18 二、 BE3能改善NMDA誘導神經細胞不正常去極化 18 三、 BE3無法改善AMPA誘導的神經細胞不正常去極化 19 四、 BE3能抑制細胞內鈣離子因Aβ、NMDA及AMPA所誘發的濃度變化 19 五、 BE3無法直接影響NMDA、AMPA受體所誘導的鈣離子流入 21 六、 BE3可降低由Aβ導致之ERK蛋白磷酸化 22 七、 BE3可降低由NMDA導致之ERK蛋白磷酸化 22 八、 BE3不會影響小鼠體重以及一些基本生理參數 23 九、 給予BE3能改善J20小鼠空間學習記憶以及辨識能力 24 (一)、巴恩斯迷宮 24 (二)、莫氏迷津水迷宮 25 (三)、新物體辨識 26 第五章 討論 27 圖表 31 圖一、BE3不影響膜電位染劑之螢光亮度 32 圖二、BE3抑制因Aβ所誘發的神經不正常去極化 34 圖三、BE3能改善NMDA誘導神經細胞不正常去極化 36 圖四、BE3無法改善AMPA誘導的神經細胞不正常去極化 38 圖五、BE3可以抑制細胞內鈣離子的濃度變化 40 圖六、BE3在第五分鐘不影響離子型麩胺酸接受器NMDA、AMPA receptor 42 圖七、BE3可降低由Aβ導致之ERK蛋白磷酸化 44 圖八、BE3可降低由NMDA導致之ERK蛋白磷酸化 46 圖九、小鼠基因型檢測以及給予BE3後的體重變化 48 圖十、BE3不影響小鼠GOT、GPT以及組織型態 50 圖十一、給予BE3能改善J20小鼠於巴恩斯迷宮之空間學習記憶能力 52 圖十二、給予BE3能改善J20小鼠於莫氏迷津水迷宮之空間學習記憶能力 54 圖十三、給予BE3能改善J20小鼠於新物體辨識之辨識認知能力 56 參考文獻 57 附錄 62 附錄一、神經細胞免疫染色 63 附錄二、β類澱粉蛋白寡聚體製備之形式 65 附錄三、黃芩素 (BE) 及黃芩素衍生物 (BE3) 之結構 67

    Alhazmi, H.A. & Albratty, M. (2022) An update on the novel and approved drugs for Alzheimer disease. Saudi Pharm J, 30, 1755-1764.
    Arndt, J.W., Qian, F., Smith, B.A., Quan, C., Kilambi, K.P., Bush, M.W., Walz, T., Pepinsky, R.B., Bussière, T., Hamann, S., Cameron, T.O. & Weinreb, P.H. (2018) Structural and kinetic basis for the selectivity of aducanumab for aggregated forms of amyloid-β. Sci Rep, 8, 6412.
    Barnes, C.A. (1979) Memory deficits associated with senescence: a neurophysiological and behavioral study in the rat. J Comp Physiol Psychol, 93, 74-104.
    Brewer, G.J. & Torricelli, J.R. (2007) Isolation and culture of adult neurons and neurospheres. Nat Protoc, 2, 1490-1498.
    Chen, C., Li, X., Gao, P., Tu, Y., Zhao, M., Li, J., Zhang, S. & Liang, H. (2015) Baicalin attenuates alzheimer-like pathological changes and memory deficits induced by amyloid β1-42 protein. Metab Brain Dis, 30, 537-544.
    Chen, G.F., Xu, T.H., Yan, Y., Zhou, Y.R., Jiang, Y., Melcher, K. & Xu, H.E. (2017) Amyloid beta: structure, biology and structure-based therapeutic development. Acta Pharmacol Sin, 38, 1205-1235.
    Cummings, J.L., Tong, G. & Ballard, C. (2019) Treatment Combinations for Alzheimer's Disease: Current and Future Pharmacotherapy Options. J Alzheimers Dis, 67, 779-794.
    Dalvi, A. (2012) Alzheimer's disease. Dis Mon, 58, 666-677.
    Dinda, B., Dinda, S., DasSharma, S., Banik, R., Chakraborty, A. & Dinda, M. (2017) Therapeutic potentials of baicalin and its aglycone, baicalein against inflammatory disorders. Eur J Med Chem, 131, 68-80.
    Esposito, Z., Belli, L., Toniolo, S., Sancesario, G., Bianconi, C. & Martorana, A. (2013) Amyloid β, glutamate, excitotoxicity in Alzheimer's disease: are we on the right track? CNS Neurosci Ther, 19, 549-555.
    Ghasemi, R., Moosavi, M., Zarifkar, A., Rastegar, K. & Maghsoudi, N. (2015) The Interplay of Akt and ERK in Aβ Toxicity and Insulin-Mediated Protection in Primary Hippocampal Cell Culture. J Mol Neurosci, 57, 325-334.
    Giraldo, E., Lloret, A., Fuchsberger, T. & Viña, J. (2014) Aβ and tau toxicities in Alzheimer's are linked via oxidative stress-induced p38 activation: protective role of vitamin E. Redox Biol, 2, 873-877.
    Grynkiewicz, G., Poenie, M. & Tsien, R.Y. (1985) A new generation of Ca2+ indicators with greatly improved fluorescence properties. J Biol Chem, 260, 3440-3450.
    Jadhav, R. & Kulkarni, Y.A. (2023) Effects of baicalein with memantine on aluminium chloride-induced neurotoxicity in Wistar rats. Front Pharmacol, 14, 1034620.
    Jin, X., Liu, M.Y., Zhang, D.F., Zhong, X., Du, K., Qian, P., Yao, W.F., Gao, H. & Wei, M.J. (2019) Baicalin mitigates cognitive impairment and protects neurons from microglia-mediated neuroinflammation via suppressing NLRP3 inflammasomes and TLR4/NF-κB signaling pathway. CNS Neurosci Ther, 25, 575-590.
    Kamat, P.K., Kalani, A., Rai, S., Swarnkar, S., Tota, S., Nath, C. & Tyagi, N. (2016) Mechanism of oxidative stress and synapse dysfunction in the pathogenesis of Alzheimer's disease: understanding the therapeutics strategies. Mol Neurobiol, 53, 648-661.
    Kelleher, R.J., 3rd, Govindarajan, A. & Tonegawa, S. (2004) Translational regulatory mechanisms in persistent forms of synaptic plasticity. Neuron, 44, 59-73.
    Kilonzo, K., Strahnen, D., Prex, V., Gems, J., van der Veen, B., Kapanaiah, S.K.T., Murthy, B.K.B., Schulz, S., Sprengel, R., Bannerman, D. & Kätzel, D. (2022) Distinct contributions of GluA1-containing AMPA receptors of different hippocampal subfields to salience processing, memory and impulse control. Transl Psychiatry, 12, 102.
    Kirvell, S.L., Esiri, M. & Francis, P.T. (2006) Down-regulation of vesicular glutamate transporters precedes cell loss and pathology in Alzheimer's disease. J Neurochem, 98, 939-950.
    Lane, C.A., Hardy, J. & Schott, J.M. (2018) Alzheimer's disease. Eur J Neurol, 25, 59-70.
    Lanooij, S.D., Drinkenburg, W., Eisel, U.L.M., van der Zee, E.A. & Kas, M.J.H. (2023) The effects of social environment on AD-related pathology in hAPP-J20 mice and tau-P301L mice. Neurobiol Dis, 187, 106309.
    Leger, M., Quiedeville, A., Bouet, V., Haelewyn, B., Boulouard, M., Schumann-Bard, P. & Freret, T. (2013) Object recognition test in mice. Nat Protoc, 8, 2531-2537.
    Lin, T.S., Tsai, H.J., Lee, C.H., Song, Y.Q., Huang, R.S., Hsieh-Li, H.M., Liang, M.C. & Lin, Y. (2017) An improved drugs screening system reveals that baicalein ameliorates the Aβ/AMPA/NMDA-Induced depolarization of neurons. J Alzheimers Dis, 56, 959-976.
    Lisman, J., Yasuda, R. & Raghavachari, S. (2012) Mechanisms of CaMKII action in long-term potentiation. Nat Rev Neurosci, 13, 169-182.
    Matsuo, N., Yamasaki, N., Ohira, K., Takao, K., Toyama, K., Eguchi, M., Yamaguchi, S. & Miyakawa, T. (2009) Neural activity changes underlying the working memory deficit in alpha-CaMKII heterozygous knockout mice. Front Behav Neurosci, 3, 20.
    Morris, R. (1984) Developments of a water-maze procedure for studying spatial learning in the rat. J Neurosci Methods, 11, 47-60.
    Mucke, L., Masliah, E., Yu, G.Q., Mallory, M., Rockenstein, E.M., Tatsuno, G., Hu, K., Kholodenko, D., Johnson-Wood, K. & McConlogue, L. (2000) High-level neuronal expression of abeta 1-42 in wild-type human amyloid protein precursor transgenic mice: synaptotoxicity without plaque formation. J Neurosci, 20, 4050-4058.
    Owen, R.T. (2016) Memantine and donepezil: a fixed drug combination for the treatment of moderate to severe Alzheimer's dementia. Drugs Today (Barc), 52, 239-248.
    Rai, S.N., Dilnashin, H., Birla, H., Singh, S.S., Zahra, W., Rathore, A.S., Singh, B.K. & Singh, S.P. (2019) The Role of PI3K/Akt and ERK in Neurodegenerative Disorders. Neurotoxicity Research, 35, 775-795.
    Revett, T.J., Baker, G.B., Jhamandas, J. & Kar, S. (2013) Glutamate system, amyloid ß peptides and tau protein: functional interrelationships and relevance to Alzheimer disease pathology. J Psychiatry Neurosci, 38, 6-23.
    Sakaguchi, Y. & Sakurai, Y. (2020) Left-right functional difference of the rat dorsal hippocampus for short-term memory and long-term memory. Behav Brain Res, 382, 112478.
    Sanderson, D.J., McHugh, S.B., Good, M.A., Sprengel, R., Seeburg, P.H., Rawlins, J.N. & Bannerman, D.M. (2010) Spatial working memory deficits in GluA1 AMPA receptor subunit knockout mice reflect impaired short-term habituation: evidence for Wagner's dual-process memory model. Neuropsychologia, 48, 2303-2315.
    Shipton, O.A., El-Gaby, M., Apergis-Schoute, J., Deisseroth, K., Bannerman, D.M., Paulsen, O. & Kohl, M.M. (2014) Left-right dissociation of hippocampal memory processes in mice. Proc Natl Acad Sci U S A, 111, 15238-15243.
    Song, Z., He, C., Yu, W., Yang, M., Li, Z., Li, P., Zhu, X., Xiao, C. & Cheng, S. (2022) Baicalin Attenuated Aβ (1-42)-Induced Apoptosis in SH-SY5Y Cells by Inhibiting the Ras-ERK Signaling Pathway. Biomed Res Int, 2022, 9491755.
    Spitzer, P., Schieb, H., Kamrowski-Kruck, H., Otto, M., Chiasserini, D., Parnetti, L., Herukka, S.K., Schuchhardt, J., Wiltfang, J. & Klafki, H.W. (2011) Evidence for elevated cerebrospinal fluid ERK1/2 levels in Alzheimer dementia. Int J Alzheimers Dis, 2011, 739847.
    Wang, M., Qiu, S. & Qin, J. (2019) Baicalein induced apoptosis and autophagy of undifferentiated thyroid cancer cells by the ERK/PI3K/Akt pathway. Am J Transl Res, 11, 3341-3352.
    Wang, Z.Y., Jiang, Z.M., Xiao, P.T., Jiang, Y.Q., Liu, W.J. & Liu, E.H. (2020) The mechanisms of baicalin ameliorate obesity and hyperlipidemia through a network pharmacology approach. Eur J Pharmacol, 878, 173103.
    Wright, A.L., Zinn, R., Hohensinn, B., Konen, L.M., Beynon, S.B., Tan, R.P., Clark, I.A., Abdipranoto, A. & Vissel, B. (2013) Neuroinflammation and neuronal loss precede Aβ plaque deposition in the hAPP-J20 mouse model of Alzheimer's disease. PLoS One, 8, e59586.
    Wu, P.F., Bhore, N., Lee, Y.L., Chou, J.Y., Chen, Y.W., Wu, P.Y., Hsu, W.M., Lee, H., Huang, Y.S., Lu, P.J. & Liao, Y.F. (2020) Phosphatidylinositol-4-phosphate 5-kinase type 1α attenuates Aβ production by promoting non-amyloidogenic processing of amyloid precursor protein. FASEB J, 34, 12127-12146.
    Xie, X.M., Hao, J.J., Shi, J.Z., Zhou, Y.F., Liu, P.F., Wang, F., Zheng, X.M., Yu, X.Y., Wang, C.C., Yan, Y., Du, G.H., Song, J.K., He, Y.Y. & Pang, X.B. (2023) Baicalein ameliorates Alzheimer's disease via orchestration of CX3CR1/NF-κB pathway in a triple transgenic mouse model. Int Immunopharmacol, 118, 109994.
    Yamasaki, N., Maekawa, M., Kobayashi, K., Kajii, Y., Maeda, J., Soma, M., Takao, K., Tanda, K., Ohira, K., Toyama, K., Kanzaki, K., Fukunaga, K., Sudo, Y., Ichinose, H., Ikeda, M., Iwata, N., Ozaki, N., Suzuki, H., Higuchi, M., Suhara, T., Yuasa, S. & Miyakawa, T. (2008) Alpha-CaMKII deficiency causes immature dentate gyrus, a novel candidate endophenotype of psychiatric disorders. Mol Brain, 1, 6.
    Yokoyama, M., Kobayashi, H., Tatsumi, L. & Tomita, T. (2022) Mouse Models of Alzheimer's Disease. Front Mol Neurosci, 15, 912995.
    Zhang, H., Wei, W., Zhao, M., Ma, L., Jiang, X., Pei, H., Cao, Y. & Li, H. (2021) Interaction between Aβ and Tau in the Pathogenesis of Alzheimer's Disease. Int J Biol Sci, 17, 2181-2192.

    無法下載圖示 電子全文延後公開
    2029/04/10
    QR CODE