臺灣博碩士論文加值系統

癲癇為常見神經性疾病，而導致癲癇的因素則是由於腦中神經細胞不正常放電。除此之外，臨床上越來越多癲癇病患容易伴隨著情緒障礙等合併症，像是罹患焦慮及憂鬱。另外在過去的研究提到，一個細胞自噬的損傷則會導致癲癇的好發並造成腦損傷。然而，細胞自噬是否會導致癲癇引起的情緒障礙仍然不清楚。儘管如此，抗癲癇藥物（AEDs）是臨床上常用來治療癲癇的方式，但仍有30％的癲癇患者未得到充分的控制。此外，抗癲癇藥物產生的副作用，再加上過去研究提及長期服用抗癲癇藥物，則會造成癲癇患者中維生素卻乏，都會影響患者服用AEDs意願，進而加劇癲癇的疾病。此外，越來越多的證據表明，維生素補充劑可以誘導自噬，並在許多神經系統疾病的預防和治療中發揮潛在作用。因此，我們欲探討補充維生素輔助較低劑量的AEDs是否能對於癲癇以及癲癇誘發之情緒疾病更有療效，藉由透過細胞自噬的機制。然而，在實驗結果顯示，給予綜合維生素輔助較低劑量的抗癲癇藥物，對於輕度和重度癲癇以及癲癇所誘發之情緒疾病是更具有療效。接著，我們進一步去測量杏仁核及海馬迴中細胞自噬的表現量，可以看到在基因層面，綜合維生素輔助較低劑量的抗癲癇藥物在杏仁核中，更顯著提升atg5、lc3a和降低p62; 另外在海馬迴中則是更增加becn1、atg5、lc3a和lc3b mRNA得表現量。但再給予了細胞自噬活化劑(rapamycin)及抑制劑(chloroquine)後，是看到細胞自噬活化劑可以去緩解輕度癲癇，而給予了細胞自噬抑制劑則是會去緩解重度癲癇。因此綜合維生素輔助較低劑量的抗癲癇藥物，可以去緩解癲癇及其誘發之情緒疾病，並且是透過促進autophagy在輕度癲癇是緩解，而抑制autophagy來緩解重度癲癇。

Epilepsy is a common neurological disorders and is characterized by a hyperexcitatory of the brain neurons. Recently, it is suggested that mood disorder (such as anxiety and depression) is observed in patients suffering from epilepsy. Furthermore, previous studies revealed that autophagy impairment contributed to epilepsy-induced brain damage. However, autophagy contributes to epilepsy-induced mood disorders still remains unclear. Although the antiepileptic drugs (AEDs) are widely and effectively used, about 30% epilepsy patients are still not adequately controlled. Moreover, AEDs-mediated side effects disturb the willing to take AEDs and reduce the patient’s quality of life. Besides, increasing evidences have revealed that vitamin supplementary could induce autophagy and play a potential role in prophylactic and therapeutic in many neurological disorders. So we hypothesized that the supplementary of vitamins combined with AEDs could be a potential strategy for treating mood disorders in an epileptic animal model via an activation of autophagy mechanism. Our results showed that a reduction of seizure severity and anxiety were found in rats receiving combinational vitamin supplementation and AEDs. Moreover, we found that AEDs application increased the mRNA levels of atg5、lc3a and decreased the mRNA levels of p62 in amygdala of seizure rats receiving vitamins supplementation. On the other hand, we found that increased of becn1、atg5、lc3a and lc3b mRNA levels in hippocampus. Furthermore, we applied the autophagy activator rapamycin and autophagy inhibitor chloroquine in seizure rats receiving vitamins supplementation. We found that rapamycin could alleviate mild epilepsy, and chloroquine could reduce the severe epilepsy in seizure rats receiving vitamins supplementation. Therefore, it suggested that combinational vitamin supplementation and low dose AEDs could alleviate seizure and relevant anxiety in rats via an activation and inhibition of autophagy.

中文摘要 I
Abstract II
致謝 III
目錄 IV
圖表目錄 VI
縮寫 (Abbreviation) VIII
第一章 緒論 (Introduction) 1
一、 癲癇 (Epilepsy) 1
三、 抗癲癇藥物（Anti-epileptic drugs; AEDs） 2
四、 維生素 (Vitamins) 3
五、 細胞自噬(Autophagy) 5
第二章 材料與方法 (Materials And Methods) 7
一、 實驗動物 7
二、 顳葉型癲癇症動物模式之建立 7
三、 實驗設計 8
四、 動物實驗藥品配製 9
五、 動物行為實驗（Animal behavior test） 9
六、 動物全身灌流與腦組織製備 10
七、 即時聚合酶鏈鎖反應（Quantitative real time polymerase chain reaction, rt-PCR) 11
八、 西方點墨法（Western blot） 12
九、 酵素連結免疫吸附分析法 (Enzyme-linked immunosorbent assay, ELISA)測定腦組織及血漿中Vitamin E 及CoQ10之含量 15
十、 統計分析 16
第三章 結果 (Results) 17
一、 顳葉型癲癇症動物模式之建立 17
二、 癲癇大鼠誘發之情緒障礙 17
三、 抗癲癇藥物對於癲癇及癲癇所誘發的情緒障礙之療效 17
四、 癲癇大鼠中及給予抗癲癇藥物後，細胞自噬在杏仁核及海馬迴之角色 18
五、 餵食不同週數綜合維生素對於癲癇之療效 19
六、 綜合維生素輔助較低劑量不同種抗癲癇藥物之療效 20
七、 綜合維生素輔助較低劑量不同種抗癲癇藥物對於癲癇誘發情緒障障礙之療效 21
八、 癲癇大鼠中給予綜合維生素輔助較低劑量不同種抗癲癇藥物，其細胞自噬在杏仁核及海馬迴之角色 22
第四章 討論 (Disussion) 24
第五章 研究重要性 (Significance of the study) 25
第六章 參考文獻 27
第七章 結果圖 (Figures) 31

Altunsoy, N., et al. (2018). Exploring the relationship between vitamin D and mania: correlations between serum vitamin D levels and disease activity. Nordic journal of psychiatry 72(3): 221-225.

Dong, Y., et al. (2013). Ascorbic acid ameliorates seizures and brain damage in rats through inhibiting autophagy. Brain research 1535: 115-123.

French, J. A. and T. A. Pedley (2008). Initial Management of Epilepsy. New England Journal of Medicine 359(2): 166-176.

Gan, J., et al. (2015). An evaluation of the links between microRNA, autophagy, and epilepsy. Reviews in the neurosciences 26(2): 225-237.

Higgins, J. P., et al. (2010). Energy beverages: content and safety. Mayo clinic proceedings, Elsevier.

Hollo, A., et al. (2014). Epilepsy and vitamin D. Int J Neurosci 124(6): 387-393.

Ihara, Y., et al. (2016). Retigabine, a Kv7. 2/Kv7. 3-channel opener, attenuates drug-induced seizures in knock-in mice harboring Kcnq2 mutations. PLoS One 11(2): e0150095.

Leon, A. C., et al. (2012). Antiepileptic drugs for bipolar disorder and the risk of suicidal behavior: a 30-year observational study. American Journal of Psychiatry 169(3): 285-291.

Lumley, J., et al. (2001). Periconceptional supplementation with folate and/or multivitamins for preventing neural tube defects. Cochrane database of systematic reviews(3).

Manon‐Espaillat, R., et al. (1991). Antiepileptic drug intoxication: factors and their significance. Epilepsia 32(1): 96-100.

McMahon, J., et al. (2012). Impaired autophagy in neurons after disinhibition of mammalian target of rapamycin and its contribution to epileptogenesis. Journal of Neuroscience 32(45): 15704-15714.

Mula, M. and J. W. Sander (2007). Negative effects of antiepileptic drugs on mood in patients with epilepsy. Drug Safety 30(7): 555-567.