頸部脊髓損傷後呼吸功能缺陷是病患主要死亡原因，由於直接破壞橫膈運動神經核導致無法正常支配橫膈肌。目前頸部脊髓挫傷模型無法有效模擬呼吸功能缺陷的臨床狀況，因此本論文的首要目的在建立與臨床病徵相符的中段頸部脊髓挫傷模型。先前研究結果顯示頸部第2節半切動物可阻斷延腦傳遞至橫膈運動神經核路徑，並顯著減弱肺迷走神經介導之肺化學反射，而本論文使用中段頸部脊髓挫傷模型可保留部分延腦呼吸傳遞路徑但顯著破壞橫膈運動神經核，有鑑於兩種模型損傷型態有別，因此本論文再進一步探討中段頸部脊髓挫傷模型肺迷走神經介導的呼吸反射變化。將成年雄性Sprague Dawley大鼠在頸部第3-4節脊髓中線進行不同高度撞擊(中度：6.25 mm、重度：12.5 mm)的挫傷手術，於術後1天及1、2、4、8週使用全體腔呼吸測量系統(whole body barometric plethysmography)測量清醒動物於呼吸空氣及高二氧化碳氣體 (7%二氧化碳，21%氧氣，72%氮氣)的呼吸型態變化。損傷後3天、2週與8週由頸靜脈注射辣椒素(1.5 µg/kg)或增加呼氣末壓力(9 cmH2O)觀察肺化學反射及肺脹大反射變化。結果顯示損傷組別在損傷後1天相較於未損傷組別潮氣容積顯著下降、呼吸頻率加快，在刺激氣體下潮氣容積及呼吸頻率皆低於未損傷組別，在損傷後8週各組別呼吸頻率無明顯差異，但損傷組潮氣容積仍然明顯低於未損傷組別。辣椒素刺激可顯著延長未損傷組別呼氣時間，但在損傷後3天挫傷組別呼氣延長反應顯著減弱。肺脹大反射可引起未損傷及中度挫傷組別呼氣時間延長，但重度挫傷組別呼氣延長反應顯著衰弱。損傷後2週及8週肺化學反射及肺脹大反射在所有組別皆無明顯差異。綜合以上結果顯示頸部第3-4節脊髓中線挫傷動物可導致呼吸功能長期缺陷，由肺迷走神經介導的呼吸反射在損傷急性期產生短暫減弱反應至損傷後2-8週則可逐漸恢復。

Cervical spinal cord injury is associated with high mortality and morbidity due to respiratory dysfunction. The current cervical spinal cord injury model can’t simulate the clinical respiratory syndrome. Accordingly, the first aim of the present study is to establish a mid-cervical spinal cord contusion animal model. Previous studies have shown that vagal-mediated pulmonary chemoreflex was attenuated following acute and chronic unilateral C2 hemisection. The second aim of this study is to examine whether vagal-mediated respiratory reflexes was altered following cervical contusion. Moderate and severe contusion was induced at midline C3-4 spinal cord in adult Sprague Dawley rats. The ventilatory behaviors of unanesthetized animals during compressed air breathing and hypercapnic challenge were evaluated by the whole body barometric plethysmography at 1 day and 1, 2, 4, and 8 weeks post-injury. The pulmonary chemoreflex induced by intra-jugular capsaicin injection and Hering-Breuer inflation reflex induced by increasing positive end-expired pressure were evoked at 3 days, 2 or 8 weeks post-surgery. The results showed that tidal volume of unanesthetized rats was decreased in severe animals compared with uninjured animals at all-time points after injury. At 3 days post-injury, capsaicin administration induced a significant apnea, but this response was greatly attenuated in both contused groups. Increased positive end-expired pressure caused an increase in the expiratory duration in uninjured and moderate contused animals but not severe contused animals. At 2 and 8 weeks post-injury, the respiratory reflex induced by capsaicin or lung inflation was similar between uninjured and contused animals. These data suggested that mid-cervical contusion injury can induce a long-term respiratory impairment, and lung vagal-mediated respiratory reflexes were transiently attenuated.

論文審定書 i
中文摘要 ii
Abstract iii
圖次 vi
表次 viii
第一章 前言 1
1-1 頸部脊髓損傷對於呼吸功能影響 1
1-2 迷走傳入神經對於呼吸的調控作用 4
1-3 脊髓損傷後治療方向 5
1-4 目的 7
第二章 材料與方法 9
2-1 實驗組別 9
2-2 頸部脊髓挫傷手術 9
2-3 清醒動物呼吸測量 10
2-4 麻醉動物呼吸、心跳與血壓測量 11
2-5 麻醉動物血液參數測量 11
2-6 辣椒素配置 11
2-7 肺脹大刺激裝置 12
2-8 實驗計畫表 12
2-9 數據分析與統計 14
第三章 結果 17
3-1 動物於頸部脊髓挫傷後的存活狀況與體重變化 17
3-2 頸部脊髓挫傷後清醒動物的呼吸型態 17
3-3 頸部脊髓挫傷後麻醉動物的呼吸型態、心跳及血壓 18
3-4 辣椒素誘發之肺化學反射於頸部脊髓損傷後的變化 18
3-5 肺脹大誘發肺反射於頸部脊髓損傷的影響 19
3-6 切除雙側迷走神經對於呼吸功能與心跳和血壓影響 20
3-7 切除雙側迷走神經在辣椒素誘發肺化學反射於頸部脊髓損傷後的變化 21
3-8 切除雙側迷走神經在肺脹大誘發肺反射於頸部脊髓損傷的影響 21
3-9 組織學 21
3-10 逆向神經追蹤劑標定橫膈運動神經元 22
第四章 討論 23
4-1 方法討論 23
4-2 清醒動物與麻醉動物基礎值下呼吸功能差異 24
4-3 頸部脊髓損傷後肺化學反射及肺脹大反射的變化 25
4-4 橫膈運動神經元數量對於損傷後呼吸功能的影響 28
4-5 結論 28
第五章 圖與說明 30
第六章 表與說明 53
第七章 參考文獻 60
第八章 附錄 66

Allbutt, H. N., P. J. Siddall and K. A. Keay (2007). "Contusive spinal cord injury evokes localized changes in NADPH-d activity but extensive changes in Fos-like immunoreactivity in the rat." J Anat 211(3): 352-370.
Alvarez-Argote, S., H. M. Gransee, J. C. Mora, J. M. Stowe, A. J. Jorgenson, G. C. Sieck and C. B. Mantilla (2016). "The Impact of Midcervical Contusion Injury on Diaphragm Muscle Function." J Neurotrauma 33(5): 500-509.
Baussart, B., J. C. Stamegna, J. Polentes, M. Tadie and P. Gauthier (2006). "A new model of upper cervical spinal contusion inducing a persistent unilateral diaphragmatic deficit in the adult rat." Neurobiol Dis 22(3): 562-574.
Bergren, D. R. and D. F. Peterson (1993). "Identification of vagal sensory receptors in the rat lung: are there subtypes of slowly adapting receptors?" J Physiol 464: 681-698.
Berlly, M. and K. Shem (2007). "Respiratory management during the first five days after spinal cord injury." J Spinal Cord Med 30(4): 309-318.
Bolser, D. C., S. C. Jefferson, M. J. Rose, N. J. Tester, P. J. Reier, D. D. Fuller, P. W. Davenport and D. R. Howland (2009). "Recovery of airway protective behaviors after spinal cord injury." Respir Physiol Neurobiol 169(2): 150-156.
Carr, M. J. and B. J. Undem (2003). "Bronchopulmonary afferent nerves." Respirology 8(3): 291-301.
Carr, M. J. and B. J. Undem (2003). "Pharmacology of vagal afferent nerve activity in guinea pig airways." Pulm Pharmacol Ther 16(1): 45-52.
Choi, H., W. L. Liao, K. M. Newton, R. C. Onario, A. M. King, F. C. Desilets, E. J. Woodard, M. E. Eichler, W. R. Frontera, S. Sabharwal and Y. D. Teng (2005). "Respiratory abnormalities resulting from midcervical spinal cord injury and their reversal by serotonin 1A agonists in conscious rats." J Neurosci 25(18): 4550-4559.
Chuaychoo, B., M. G. Lee, M. Kollarik and B. J. Undem (2005). "Effect of 5-hydroxytryptamine on vagal C-fiber subtypes in guinea pig lungs." Pulm Pharmacol Ther 18(4): 269-276.
Coleridge, J. C. and H. M. Coleridge (1984). "Afferent vagal C fibre innervation of the lungs and airways and its functional significance." Rev Physiol Biochem Pharmacol 99: 1-110.
Davies, A., M. Dixon, D. Callanan, A. Huszczuk, J. G. Widdicombe and J. C. Wise (1978). "Lung reflexes in rabbits during pulmonary stretch receptor block by sulphur dioxide." Respir Physiol 34(1): 83-101.
Delpierre, S., C. Grimaud, Y. Jammes and N. Mei (1981). "Changes in activity of vagal bronchopulmonary C fibres by chemical and physical stimuli in the cat." J Physiol 316: 61-74.
DeVivo, M. J. (2016). "Spinal Cord Injury (SCI) Facts and Figures at a Glance." J Spinal Cord Med 39(3): 370-371.
Dobbins, E. G. and J. L. Feldman (1994). "Brainstem network controlling descending drive to phrenic motoneurons in rat." J Comp Neurol 347(1): 64-86.
Dutta, A., A. Akella and S. B. Deshpande (2013). "A study to investigate capsaicin-induced pressure response in vagotomized rats." Indian J Pharmacol 45(4): 365-370.
Ellenberger, H. H. (1988). "Monosynaptic transmission of respiratory drive to phrenic motoneurons from brainstem bulbospinal neurons in rats." J Comp Neurol 269(1): 47-57.
Ellenberger, H. H., J. L. Feldman and H. G. Goshgarian (1990). "Ventral respiratory group projections to phrenic motoneurons: electron microscopic evidence for monosynaptic connections." J Comp Neurol 302(4): 707-714.
Forgione, N., S. K. Karadimas, W. D. Foltz, K. Satkunendrarajah, A. Lip and M. G. Fehlings (2014). "Bilateral contusion-compression model of incomplete traumatic cervical spinal cord injury." J Neurotrauma 31(21): 1776-1788.
Fuller, D. D., N. J. Doperalski, B. J. Dougherty, M. S. Sandhu, D. C. Bolser and P. J. Reier (2008). "Modest spontaneous recovery of ventilation following chronic high cervical hemisection in rats." Exp Neurol 211(1): 97-106.
Fuller, D. D., A. G. Zabka, T. L. Baker and G. S. Mitchell (2001). "Phrenic long-term facilitation requires 5-HT receptor activation during but not following episodic hypoxia." J Appl Physiol (1985) 90(5): 2001-2006; discussion 2000.
Golder, F. J., P. W. Davenport, R. D. Johnson, P. J. Reier and D. C. Bolser (2005). "Augmented breath phase volume and timing relationships in the anesthetized rat." Neurosci Lett 373(2): 89-93.
Golder, F. J., D. D. Fuller, P. W. Davenport, R. D. Johnson, P. J. Reier and D. C. Bolser (2003). "Respiratory motor recovery after unilateral spinal cord injury: eliminating crossed phrenic activity decreases tidal volume and increases contralateral respiratory motor output." J Neurosci 23(6): 2494-2501.
Golder, F. J., D. D. Fuller, M. R. Lovett-Barr, S. Vinit, D. K. Resnick and G. S. Mitchell (2011). "Breathing patterns after mid-cervical spinal contusion in rats." Exp Neurol 231(1): 97-103.
Golder, F. J., P. J. Reier, P. W. Davenport and D. C. Bolser (2001). "Cervical spinal cord injury alters the pattern of breathing in anesthetized rats." J Appl Physiol (1985) 91(6): 2451-2458.
Goshgarian, H. G. (1984). "The ultrastructure and synaptic architecture of phrenic motor neurons in the spinal cord of the adult rat." J Neurocytol 13(1): 85-109.
Hong, J. L., K. Kwong and L. Y. Lee (1997). "Stimulation of pulmonary C fibres by lactic acid in rats: contributions of H+ and lactate ions." J Physiol 500 ( Pt 2): 319-329.
Hoy, K. C., Jr. and W. J. Alilain (2015). "Acute theophylline exposure modulates breathing activity through a cervical contusion." Exp Neurol 271: 72-76.
Kalderon, N., M. Muruganandham, J. A. Koutcher and M. Potuzak (2007). "Therapeutic strategy for acute spinal cord contusion injury: cell elimination combined with microsurgical intervention." PLoS One 2(6): e565.
Kalia, M. and D. Richter (1985). "Morphology of physiologically identified slowly adapting lung stretch receptor afferents stained with intra-axonal horseradish peroxidase in the nucleus of the tractus solitarius of the cat. II. An ultrastructural analysis." J Comp Neurol 241(4): 521-535.
Kanamaru, A., T. Mutoh, R. Nishimura, N. Sasaki, M. Kuwahara and H. Tsubone (2001). "Respiratory and cardiovascular reflexes elicited by nasal instillation of capsaicin to anesthetized, spontaneously breathing dogs." J Vet Med Sci 63(4): 439-443.
Knowlton, G. C. and M. G. Larrabee (1946). "A unitary analysis of pulmonary volume receptors." Am J Physiol 147: 100-114.
Kubin, L., H. Kimura and R. O. Davies (1991). "The medullary projections of afferent bronchopulmonary C fibres in the cat as shown by antidromic mapping." J Physiol 435: 207-228.
Lane, M. A., K. Z. Lee, K. Salazar, B. E. O'Steen, D. C. Bloom, D. D. Fuller and P. J. Reier (2012). "Respiratory function following bilateral mid-cervical contusion injury in the adult rat." Exp Neurol 235(1): 197-210.
Lee, K. Z. and Y. S. Chang (2014). "Recovery of the pulmonary chemoreflex and functional role of bronchopulmonary C-fibers following chronic cervical spinal cord injury." J Appl Physiol (1985) 117(10): 1188-1198.
Lee, K. Z., M. S. Sandhu, B. J. Dougherty, P. J. Reier and D. D. Fuller (2010). "Influence of vagal afferents on supraspinal and spinal respiratory activity following cervical spinal cord injury in rats." J Appl Physiol (1985) 109(2): 377-387.
Lin, R. L., Y. J. Lin, M. J. Geer, R. Kryscio and L. Y. Lee (2013). "Pulmonary chemoreflex responses are potentiated by tumor necrosis factor-alpha in mice." J Appl Physiol (1985) 114(11): 1536-1543.
Lin, Y. S. and L. Y. Lee (2002). "Stimulation of pulmonary vagal C-fibres by anandamide in anaesthetized rats: role of vanilloid type 1 receptors." J Physiol 539(Pt 3): 947-955.
Liu, B. Y., Y. J. Lin, H. F. Lee, C. Y. Ho, T. Ruan and Y. R. Kou (2015). "Menthol suppresses laryngeal C-fiber hypersensitivity to cigarette smoke in a rat model of gastroesophageal reflux disease: the role of TRPM8." J Appl Physiol (1985) 118(5): 635-645.
Mansoor, J. K., D. M. Hyde and E. S. Schelegle (1997). "Contribution of vagal afferents to breathing pattern in rats with lung fibrosis." Respir Physiol 108(1): 45-61.
Mutoh, T., J. P. Joad and A. C. Bonham (2000). "Chronic passive cigarette smoke exposure augments bronchopulmonary C-fibre inputs to nucleus tractus solitarii neurones and reflex output in young guinea-pigs." J Physiol 523 Pt 1: 223-233.
Navarrete-Opazo, A., S. Vinit, B. J. Dougherty and G. S. Mitchell (2015). "Daily acute intermittent hypoxia elicits functional recovery of diaphragm and inspiratory intercostal muscle activity after acute cervical spinal injury." Exp Neurol 266: 1-10.
Nicaise, C., D. M. Frank, T. J. Hala, M. Authelet, R. Pochet, D. Adriaens, J. P. Brion, M. C. Wright and A. C. Lepore (2013). "Early phrenic motor neuron loss and transient respiratory abnormalities after unilateral cervical spinal cord contusion." J Neurotrauma 30(12): 1092-1099.
Nicaise, C., T. J. Hala, D. M. Frank, J. L. Parker, M. Authelet, K. Leroy, J. P. Brion, M. C. Wright and A. C. Lepore (2012). "Phrenic motor neuron degeneration compromises phrenic axonal circuitry and diaphragm activity in a unilateral cervical contusion model of spinal cord injury." Exp Neurol 235(2): 539-552.
Nicaise, C., R. Putatunda, T. J. Hala, K. A. Regan, D. M. Frank, J. P. Brion, K. Leroy, R. Pochet, M. C. Wright and A. C. Lepore (2012). "Degeneration of phrenic motor neurons induces long-term diaphragm deficits following mid-cervical spinal contusion in mice." J Neurotrauma 29(18): 2748-2760.
Oh, E. J., S. B. Mazzone, B. J. Canning and D. Weinreich (2006). "Reflex regulation of airway sympathetic nerves in guinea-pigs." J Physiol 573(Pt 2): 549-564.
Pagliardini, S., J. J. Greer, G. D. Funk and C. T. Dickson (2012). "State-dependent modulation of breathing in urethane-anesthetized rats." J Neurosci 32(33): 11259-11270.
Palecek, F., G. Sant'Ambrogio, F. B. Sant'Ambrogio and O. P. Mathew (1989). "Reflex responses to capsaicin: intravenous, aerosol, and intratracheal administration." J Appl Physiol (1985) 67(4): 1428-1437.
Pisarri, T. E., A. Jonzon, J. C. Coleridge and H. M. Coleridge (1990). "Rapidly adapting receptors monitor lung compliance in spontaneously breathing dogs." J Appl Physiol (1985) 68(5): 1997-2005.
Rabchevsky, A. G., I. Fugaccia, P. G. Sullivan, D. A. Blades and S. W. Scheff (2002). "Efficacy of methylprednisolone therapy for the injured rat spinal cord." J Neurosci Res 68(1): 7-18.
Routal, R. V. and G. P. Pal (1999). "Location of the phrenic nucleus in the human spinal cord." J Anat 195 ( Pt 4): 617-621.
Ruan, T., Q. Gu, Y. R. Kou and L. Y. Lee (2005). "Hyperthermia increases sensitivity of pulmonary C-fibre afferents in rats." J Physiol 565(Pt 1): 295-308.
Schelegle, E. S., W. F. Walby, J. K. Mansoor and A. T. Chen (2001). "Lung vagal afferent activity in rats with bleomycin-induced lung fibrosis." Respir Physiol 126(1): 9-27.
Selman, M., T. E. King and A. Pardo (2001). "Idiopathic pulmonary fibrosis: prevailing and evolving hypotheses about its pathogenesis and implications for therapy." Ann Intern Med 134(2): 136-151.
Sera, T., H. Yokota, K. Uesugi and N. Yagi (2013). "Airway distension during lung inflation in healthy and allergic-sensitised mice in vivo." Respir Physiol Neurobiol 185(3): 639-646.
Silver, W. L., L. G. Farley and T. E. Finger (1991). "The effects of neonatal capsaicin administration on trigeminal nerve chemoreceptors in the rat nasal cavity." Brain Res 561(2): 212-216.
Simard, J. M., P. G. Popovich, O. Tsymbalyuk, J. Caridi, R. P. Gullapalli, M. J. Kilbourne and V. Gerzanich (2013). "MRI evidence that glibenclamide reduces acute lesion expansion in a rat model of spinal cord injury." Spinal Cord 51(11): 823-827.
Teng, Y. D., I. Mocchetti, A. M. Taveira-DaSilva, R. A. Gillis and J. R. Wrathall (1999). "Basic fibroblast growth factor increases long-term survival of spinal motor neurons and improves respiratory function after experimental spinal cord injury." J Neurosci 19(16): 7037-7047.
Teng, Y. D., I. Mocchetti and J. R. Wrathall (1998). "Basic and acidic fibroblast growth factors protect spinal motor neurones in vivo after experimental spinal cord injury." Eur J Neurosci 10(2): 798-802.
Tsai, I. L. and K. Z. Lee (2014). "Attenuation of the pulmonary chemoreflex following acute cervical spinal cord injury." J Appl Physiol (1985) 116(7): 757-766.
Undem, B. J., B. Chuaychoo, M. G. Lee, D. Weinreich, A. C. Myers and M. Kollarik (2004). "Subtypes of vagal afferent C-fibres in guinea-pig lungs." J Physiol 556(Pt 3): 905-917.
Wang, R. and F. Xu (2006). "Postnatal development of right atrial injection of capsaicin-induced apneic response in rats." J Appl Physiol (1985) 101(1): 60-67.
Widdicombe, J. (2006). "Reflexes from the lungs and airways: historical perspective." J Appl Physiol (1985) 101(2): 628-634.
Widdicombe, J. G. (1954). "Receptors in the trachea and bronchi of the cat." J Physiol 123(1): 71-104.
Wu, J. C., Y. C. Chen, L. Liu, T. J. Chen, W. C. Huang, H. Cheng and T. P. Su (2012). "Effects of age, gender, and socio-economic status on the incidence of spinal cord injury: an assessment using the eleven-year comprehensive nationwide database of Taiwan." J Neurotrauma 29(5): 889-897.
Xu, J., W. Yang, G. Zhang, Q. Gu and L. Y. Lee (2007). "Calcium transient evoked by nicotine in isolated rat vagal pulmonary sensory neurons." Am J Physiol Lung Cell Mol Physiol 292(1): L54-61.
Zhang, G., R. L. Lin, M. Wiggers, D. M. Snow and L. Y. Lee (2008). "Altered expression of TRPV1 and sensitivity to capsaicin in pulmonary myelinated afferents following chronic airway inflammation in the rat." J Physiol 586(23): 5771-5786.
Zimmer, M. B. and H. G. Goshgarian (2007). "Spinal cord injury in neonates alters respiratory motor output via supraspinal mechanisms." Exp Neurol 206(1): 137-145.
Zimmer, M. B., K. Nantwi and H. G. Goshgarian (2007). "Effect of spinal cord injury on the respiratory system: basic research and current clinical treatment options." J Spinal Cord Med 30(4): 319-330.