臺灣博碩士論文加值系統

本研究旨在研究cinnamophilin（CINN）對於實驗鼠缺血性腦中風之神經保護特性，治療有效期限與相關機轉等之探討。近年來，CINN為本土神經科學之一熱門話題，相關研究發現CINN是一強而有效之自然抗氧化劑與自由基截取劑，且可抑制缺血後之血小板聚集，是以有潛力降低缺血後的能量流失。先前的研究已發現CINN於急性缺血性腦中風有強而有效之神經保護效果。而在本研究中，利用S.D.大白鼠進行持續性中大腦動脈縫線栓塞並以CINN治療，發現CINN是一強而有效之神經保護劑，且並無明顯不當之副作用。
我們利用大白鼠進行可逆轉之中大腦動脈縫線栓塞逐步研究探討缺血治療後之短期追蹤，評估其進一步之神經保護特性與其神經保護潛能。並於可逆轉中大腦動脈縫線栓塞後7天，應用影像處理系統計算Nissl染色腦切片之栓塞大小並評估栓塞後個別神經運動感覺功能，腦血流、腦電生理與體重回復。
由實驗結果發現，以80 mg／kg之CINN進行靜脈注射為有效反應劑量，並於持續性中大腦動脈縫線栓塞後至少2小時仍具有效神經保護。也發現在缺血120分鐘之後以CINN進行緩慢靜脈注射，藉由SSEP之結果觀察得知，在左側（大腦動脈阻塞側）的上肢與下肢之實驗前後的P1- N1比較之中，CINN治療組比HPCD控制組的振幅增加約30％，表示動作電位傳遞有不錯的恢復效果，顯示CINN加強了神經生理的結果。另外由免疫組織化學染色觀察到，在腦部缺血／再灌流之後，cortical infarction以及striatal infarction之體積有降低的現象，顯示CINN有助於灰質損傷的降低。
總而言之，本研究觀察得到CINN是無毒性的並具有持續性的神經保護，故其對於神經外科手術的計畫性腦血管栓塞以及缺血性腦中風的病人具有潛在性的益處，值得進一步的研究。

The neuroprotective properties of cinnamophilin, a strong natural antioxidant and free radical scavenger which may enhance cerebral hemodynamics, decrease platelet aggregation and lipid peroxidation, and prevent energy depletion during ischemia, will be evaluated in Sprague-Dawley rats subjected to 1-hour transient middle cerebral artery (MCA) occlusion. A series of experiments with delayed treatment of CINN was employed to examine whether CINN is neuroprotective following a short period of recovery using a model of reversible MCA occlusion which includes ischemic brain damage not only caused by grey matter but also by white matter damages. Postmortem infarct volumes will be determined by quantitative image analysis of Nissl-stained brain sections and by immunohistochemical methods. In addition, postischemic electrophysiological recovery will be evaluated. The possible mechanisms of action relevant to short-term neuroprotective efficacy by which CINN may exhibit the neuroprotective effects was evaluated in the model. The data not only extend our original findings to the ischemic-reperfusion injury but also look at prolonged outcomes, thus, may provide a potential outlook to treat patients against ischemic stroke.
Delayed intravenous administration with CINN at 120 minutes after ischemia both enhances electrophysiological and neurobehavioral outcomes and reduces cortical and striatal infarction after cerebral ischemia-reperfusion. Since CINN is noted to be lack of toxicity and has persistent neuroprotection, it is worthwhile further investigating for potential benefits for those patients who will undergo planned cerebrovascular occlusion during neurosurgical surgery and for possible applications for ischemic stroke patients.

摘要 I
Abstract II
誌謝 III
目錄 IV
表目錄 VII
圖目錄 VIII
第一章 緒論 1
1-1 腦血管疾病介紹 1
1-1-1 我國腦中風發生之現況 2
1-1-2 腦血管意外（Cerebrovascular Accidents，CVA） 3
1-2 大腦血液循環的臨床生理學與病理生理學 5
1-2-1 腦血管循環的臨床生理學 5
1-2-2 腦血管循環的病理生理學 7
1-3 電生理學的神經傳導信號監測 14
1-3-1 動作電位傳導 14
1-3-2 感覺神經病變 16
1-3-3 體感覺誘發電位（Somatosensory Evoked Potential，SSEP） 16
1-4 研究動機與目的 22
1-4-1 實驗藥物CINN 23
1-4-2 臨床缺血性中風治療藥物 24
1-4-3 研究目的 26
第二章 材料與方法 27
2-1 實驗流程 27
2-2 實驗方法 29
2-2-1 動物準備，麻醉和監控 29
2-2-2 實驗模型 29
2-2-3 LCBF監測 31
2-2-4 體感覺誘發電位（SSEP）監測 32
2-2-5 神經行為測試及體重測量 37
2-2-6 動物犧牲和局部缺血傷害的量化 40
2-3 統計分析 43
第三章 結果 44
3-1 生理參數之記錄 44
3-2 核心體溫之監控 45
3-3 局部腦皮質血流量之監測 46
3-4 體感覺誘發電位之量測 48
3-5 體重改變量及神經行為評估系統之評量 54
3-6 腦梗塞與腦水腫體積之比較 55
3-7 皮質與紋狀體腦梗塞體積大小之比較 56
第四章 討論 57
4-1 探討體感覺誘發電位量測之結果 57
4-2 就實驗程序探討影響體感覺誘發電位記錄之因素 58
4-3 神經行為的評估 60
4-4 CINN之神經保護效果 61
第五章結論與未來展望 63
5-1 結論 63
5-2 未來展望 64
參考文獻 65
自述 71

1、Newman MF, Grocott HP,Mathew JP, et al. Report of subatudy assessing the impact of neurocognitive function on quality of life 5 years after cardic surgery. Stroke. 2001；17：927-938
2、陳婉玲（民95）‧Magnolol於離體海馬迴培養缺血時的神經保護劑量及治療期效之研究‧國立成功大學醫學工程研究所碩士論文。
3、蔡宜殷（民92）‧以Melatonin治療暫時性局部腦缺血白鼠有助其電生理及神經行為之改善‧國立成功大學醫學工程研究所碩士論文。
4、邱浩彰（民88）‧神經學新知‧臺北：臺灣醫學會。
5、TOT癌症關懷照護網（民94）‧93年國人十大死因癌症連續23年蟬聯榜首‧TOT 台灣東洋癌症科學發展處：國際厚生數位科技股份有限公司。
6、林家樑（(民89）‧「膽固醇」是腦血管疾病的元兇‧臺北：亞洲醫藥網健康生活家。
7、陳仁澤 （民93）‧中風之危險因子‧臺北：行政院衛生署國民健康局。
8、張承能（民93）‧中風病患的治療與復健‧臺北：行政院衛生署國民健康局。
9、國內醫療新聞 （民93）‧漫談腦中風‧秀傳醫療體系：國際厚生數位科技股份有限公司。
10、Heart Center Online (HCO)；http://heart.healthcentersonline.com/stroke/tia.cfm
11、About，Inc.；http://adam.about.com/reports/000045.htm
12、陳俐璇（民93）‧以鎂離子治療暫時性局部腦缺血白鼠之電生理及神經行為之研究‧國立成功大學醫學工程研究所碩士論文。
13、朱復禮 （民81）‧臨床神經醫學‧臺北：合記。
14、吳進安 （民85）‧基礎神經學‧臺北：合記。
15、台灣醫學會‧台灣地區的腦梗塞‧行政院衛生署八十一年度委託研究計畫研究報告；http://www.fma.org.tw/medici al_data/taiwan19.htm#01
16、吳進安、朱復禮 （民71）‧體感覺誘發電位‧臨床醫學，9（3），218-223。
17、楊明杰、余政展、吳佳穎、吳承學、吳孟儒、李威吽BL均一、周昆達、張曜任、莊暘安、黃嘉文、詹東霖、趙又麟、趙勇全 （民87）‧Ganong醫學生理學（1）‧臺北：合記。
18、Sakatani, K., Iizuka, H., & Young, W.. Somatosensory evoked potentials in rat cerebral cortex before and after middle cerebral artery occlusion. Stroke 1990;21（1）, 124-132.
19、SiesjÖ BK. Pathophysiology and treatment of focal cerebral ischemia. Part II: Mechanisms of damage and treatment. J Neurosurg 1992; 77:337-354.
20、Traystman RJ, Kirsch JR, Koehler RC. Oxygen radical mechanisms of brain injury following ischemia and reperfusion. J Appl Physiol 1991; 71:1185-1195.
21、Kuroda S, SiesjÖ BK. Reperfusion damage following focal ischemia: pathophysiology and therapeutic windows. Clin Neurosci 1997; 4:199-212.
22、Chen JH, Wu CC, George H, Yen MH. Magnolol induces apoptosis vascular smooth muscle. Naunyn-Schmiedberg's Arch Pharmacology 2003；368：127-133
23、Zhang L, Zhang ZG, Zhang RL, et al. Postischemic（6-Hour）treatment with recombinant human tissue plasminogen activator and proteasome inhibitor PS-519 reduces infarction in a rat model of embolic focal cerebral ischemia. Stroke 2001; 32:2926-2931
24、Maeda M, Furuichi Y, Ueyama N, et al. A combined treatment with tacrolimus （FK506）and recombinant tissue plasminogen activator for thrombotic focal cerebral ischemia in rats: increased neuroprotective efficacy and extended therapeutic time window. J Cereb Blood Flow Metab 2002; 22:1205-1211.
25、Fisher M. Characterizing the target of acute stroke therapy. Stroke 1997; 28:866-872.
26、Wang YF, Tsirka SE, Strickland S, et al. Tissue plasminogen activator（t-PA）increases neuronal damage after focal cerebral ischemia in wild-type and t-PA-deficient mice. Nat Med 1998; 4:228-231.
27、Clark WM, Wissman S, Albers GW, et al. Recombinant tissue-type plasminogen activator（Alteplase）for ischemic stroke 3 to 5 hours after symptom onset. The ATLANTIS Study: a randomized controlled trial. Alteplase Thrombolysis for Acute Noninterventional Therapy in Ischemic Stroke. JAMA 1999; 282:2019-2026.
28、Asahi M, Asahi K, Wang X, et al. Reduction of tissue plasminogen activator-induced hemorrhage and brain injury by free radical spin trapping after embolic focal cerebral ischemia in rats. J Cereb Blood Flow Metab 2000; 20:452-457.
29、Lee E-Jian, Chen Hung-Yi, Lee Ming-Yang, Chen Tsung-Ying, Hsu Yun-Shang, Hu Yu-Ling, Chang Guan-Liang, Wu Tian-Shung*: Cinnamophilin reduces oxidative damage and protects against transient focal cerebral ischemia in mice. Free Radic. Biol. Med. 2005: 39(4):495-510.
30、Hsiao, G.; Teng, C.M.; Sheu, J.R.; Cheng, Y.W.; Lam, K.K.; Lee, Y.M.; Wu, T.S.; Yen, M.H. Cinnamophilin as a novel antiperoxidative cytoprotectant and free radical scavenger. Biochim. Biophys. Acta. 1525:77-88; 2001.
31、Yu, S.M.; Wu, T.S.; Teng, C.M. Pharmacological characterization of cinnamophilin, a novel dual inhibitor of thromboxane synthase and thromboxane A2 receptor. Br. J. Pharmacol. 111:906-912; 1994.
32、Yu, S.M.; Ko, F.N.; Wu, T.S.; Lee, J.Y.; Teng, C.M. Cinnamophilin, a novel thromboxane A2 receptor antagonist, isolated from Cinnamomum philippinense. Eur. J. Pharmacol. 256:85-91; 1994.
33、Su, M.J.; Chen, W.P.; Lo, T.Y.; Wu, T.S. Ionic mechanisms for the antiarrhythmic action of cinnamophilin in rat heart. J. Biomed. Sci. 6:376-86; 1999.
34、Cheng, H.T.; Chang, H. Reduction of reperfusion injury in rat skeletal muscle following administration of cinnamophilin, a novel dual inhibitor of thromboxane synthase and thromboxane A2 receptor. Thorac. Cardiovasc. Surg. 43:73-76; 1995.
35、中老年保健網‧中風防治‧中風病患的治療與復健；http://health91.bhp.doh.gov.tw/article/apoplexy9211b.htm
36、國際厚生健康園區‧腦中風病人的藥物治療‧秀傳醫療體系；http://www.24drs.com/special_report/stroke/4_4.asp
37、中老年保健網‧健康久一點‧健康殺手-腦中風；http://health91.bhp.doh.gov.tw/1-1-1.htm#top
38、台灣腦中風學會‧靜脈血栓溶解劑治療急性缺血性腦中風之一般準則；http://www.stroke.org.tw/guideline_3.htm
39、科學月刊雜誌社‧金台灣資訊事業有限公司‧圖龍文化事業股份有限公司‧新世紀生命科學課程改進計劃；http://www.dls.ym.edu.tw/lesson2/0520.htm
40、Lee, E.J.; Chen, H.Y.; Wu, T.S.; Chen, T.Y.; Ayoub, I.A.; Maynard, K.I. Acute administration of Ginkgo biloba extract（EGb 761）affords neuroprotection against permanent and transient focal cerebral ischemia in Sprague-Dawley rats. J. Neurosci. Res. 68:636-645; 2002.
41、Lee, E.J.; Wu, T.S.; Lee, M.Y.; Chen, T.Y.; Tsai, Y.Y.; Chuang, J.I.; Chang, G.L. Delayed treatment with melatonin enhances electrophysiological recovery following transient focal cerebral ischemia in rats. J. Pineal Res. 36:33-42; 2004.
42、Lee, E.J.; Lee, M.Y.; Chang, G.L.; Chen, L.H.; Hu, Y.L.; Chen, T.Y.; Wu, T.S. Delayed treatment with magnesium reduces brain infarction and improves electrophysiological recovery following transient focal cerebral ischemia in rats. J Neurosurg. 2005; 102:1085-93
43、Lee, E. J., Ayoub, I. A., Harris, F. B., Hassan, M., Ogilvy, C. S., & Maynard,K. I.. Mexiletine and magnesium independently, but not combined, protect against permanent focal cerebral ischemia in Wistar rats. J. Neurosci. Res.,1999; 58（3）, 442-448.
44、Lee, E.J.; Lee, M.Y.; Chen, H.Y.; Hsu, Y.S.; Wu, T.S.; Chen, S.T.; Chang, G.L. Melatonin attenuates gray and white matter damage in a mouse model of transient focal cerebral ischemia. J. Pineal Res. 38:42-52; 2005.
45、Pixinos G, Watson C. The rat brain in stereotaxic coordinates. Academic Press, New York, 1982.
46、Nagasawa H, Kogure K. Correlation between cerebral blood flow and histologic changes in a new rat model of middle cerebral artery occlusion. Stroke 1989; 20:1037-1043.
47、Memezawa H, Minamisawa H, Smith ML, et al. Ischemic penumbra in a model of reversible middle cerebral artery occlusion in the rat. Exp Brain Res. 1992; 89:67-78.
48、Hu, Y., Luk, K. D. K., Lu, W. W., Holmes, A., & Leong, J. C. Y.. Prevention of spinal cord injury with time-frequency analysis of evoked potentials: An experimental study. J. Neurology Neurosurgery and Psychiatry, 2001; 71（6）, 732-740.
49、Belayev, L., Alonso, O. F., Busto, R., Zhao, W., & Ginsberg, M. D.. Middle cerebral artery occlusion in the rat by intraluminal suture. Neurological and pathological evaluation of an improved model. Stroke 1996, 27, 1616-1623.
50、Clark, W. M., Rinker, L. G., Lessov, N. S., Hazel, K., Hill, J. K., Stenzel-Poore, M., & Eckenstein, F.. Lack of interleukin-6 expression is not protective against focal central nervous system ischemia. Stroke 2000, 31（7）, 1715-1720.
51、Blazevich, L.. Voltage-gated calcium channel. Continental United States: 2004; Sigma-Aldrich.
52、Colon, E. J., & Visser, S. L.. Evoked potential manual: A practical guide to clinical applications. Boston: 1990; Nijhoff.
53、Yeleswaram K, McLaughlin LG, Knipe JO, Schabdach D. Pharmacokinetics and oral bioavailability of exogenous melatonin in preclinical animal models and clinical implications. J Pineal Res 1997; 22:45-51.
54、Bolay H, Dalkara T. Mechanisms of motor dysfunction after transient MCA occlusion: persistent transmission failure in cortical synapses is a major determinant. Stroke 1998; 29:1988-1994.
55、Whishaw IQ, O'Connor WT, Dunnett SB. The contributions of motor cortex, nigrostriatal dopamine and caudate-putamen to skilled forelimb use in the rat. Brain 1986; 109:805-843.
56、Nakano S, Kogure K, Fujikura H. Ischemia-induced slowly progressive neuronal damage in the rat brain. Neuroscience 1990; 38:115-124.
57、Du C, Hu R, Csernansky CA, Hsu CY, Choi DW. Very delayed infarction after mild focal cerebral ischemia: a role for apoptosis? J Cereb Blood Flow Metab 1996; 16:195-201