臺灣博碩士論文加值系統

本研究將探討番椒晶素衍生物GLNVA與SNA對於天竺鼠離體氣管平滑肌之作用及機轉，並觀察其對於麻醉中天竺鼠呼吸道的作用。在離體實驗中發現，GLNVA (0.01-10 uM) 以及SNA (0.01-100 uM)可使天竺鼠離體氣管產生濃度依賴性的收縮作用，且此種作用能因前處理capsazepine (10 uM)， ruthenium red (10 uM)， theophylline (10 uM)，或者是破壞感覺神經所抑制。同時也發現，累積投予capsaicin (0.01-10 uM)後，加入carbachol 1 uM引起一個收縮後再投予較高濃度之GLNVA 5 uM時，對於呼吸道平滑肌反而產生鬆弛作用。而這個鬆弛作用能完全的被indomethacin (10 uM) 所抑制，也能被TEA (10-2 M) ， glibenclamide (10 uM)，theophylline (10 uM)， 8-PT (adenosine A1 receptor antagonist; 10 uM) 以及 DMPX (adenosine A2 receptor antagonist; 10 uM)部份抑制。若每隻連續7天以GLNVA 與SNA (105.6 mg)，經由頸部皮下注射投予天竺鼠，GLNVA能顯著的抑制ovalbumin (10 uM) 或capsaicin (0.01-10 uM) 所引起的氣管平滑肌之收縮作用，而SNA在抑制capsaicin的作用較小以至於因為去敏化作用的不夠而無法抑制ovalbumin所引起的氣管平滑肌之收縮作用。在活體實驗方面，每隻利用頸部皮下注射長期投予GLNVA 與SNA (105.6 mg的劑量) 前處理過的天竺鼠，麻醉後頸靜脈插管給予capsaicin (100 mg/kg)，GLNVA能顯著抑制capsaicin所引起的呼吸急促，而SNA的抑制作用較小。另外也利用物質P螢光免疫組織化學法觀察天竺鼠呼吸道的SP-like神經纖維免疫反應，結果發現capsaicin及GLNVA都能減少天竺鼠呼吸道感覺神經內的SP-like神經纖維免疫反應。從天竺鼠呼吸道的實驗得到了一個結論，就是capsaicin，GLNVA以及SNA能在感覺神經引起一個化學性的去敏化作用，而且也能利用長期的治療來降低呼吸道對於過敏原所引起的過敏性反應，這可能是因為破壞了感覺神經C-fiber所致。另外GLNVA也能活化ATP-sensitive K+ channel、adenosine receptor以及釋放出內生性的prostanoids而引起呼吸道平滑肌鬆弛作用。

The effects of glyceryl nonivamide (GLNVA) and sodium nonivamide acetate (SNA) on isolated tracheal smooth muscle of guinea-pig in vitro and the desensitizing effect of GLNVA and SNA in anesthetized guinea-pig airways in vivo were first investigated. Both GLNVA (0.01-10 uM) and SNA (0.01-100 uM) produced concentration-dependent contraction in the isolated guinea-pig trachea, and this effect were inhibited by pretreatment with capsazepine (10 uM), ruthenium red (10 uM), theophylline (10 uM), or sensory denervation, respectively. On the other hand, at higher concentration, GLNVA 5 uM could produce relaxation on carbachol-induced contracted tracheal smooth muscle after sensory denervation. Moreover, this relaxation effect was completely blocked by indomethacin (10 uM) and partially blocked by TEA (tetraethylammonium, non-selective potassium channel blocker, 10-2 M), glibenclamide (10 uM), theophylline (10 uM), 8-PT (8-phenyltheophylline, adenosine A1 receptor antagonist, 10 uM) and DMPX (3,7-dimethyl-1-propargylxanthine, adenosine A2 receptor antagonist, 10 uM) . After long-term pretreatment of GLNVA and SNA (105.6 mg, 7 days, s.c.), GLNVA significantly inhibited ovalbumin- or capsaicin-induced tracheal smooth muscle contraction, but SNA did not. In anesthetized guinea-pig, long term treatment of GLNVA and SNA, also significantly inhibited capsaicin-induced pulmonary dysfunction. The distribution of SP-like immunoreactive nerves in guinea pig airway was also displayed in this study. In the present study we find that capsaicin and the nonpungent capsaicin analogue GLNVA can deplete the SP from sensory nerve in guinea pig trachea. In conclusion, the present data indicated that, capsaicin, GLNVA and SNA caused chemical desensitization of sensory nerve in guinea-pig tracheal smooth muscle and renders them insensitive to further exposure of capsaicin. The tracheal hyperresponsivess of guinea-pig induced by ovalbumin were attenuated following chronic treatment with capsaicin, GLNVA and SNA which destroys sensory C-fiber. GLNVA induced- relaxation effect was mediated via the activation of ATP-sensitive K+ channel opening, adenosine receptor modulation and release of endogenous prostanoids of the smooth muscle.

1. 摘要 ------------------------------------------------------------1
2. 英文摘要 ------------------------------------------------------------3
3. 縮寫表 ------------------------------------------------------------5
4. 緒論 ------------------------------------------------------------6
5. 研究材料 -----------------------------------------------------------10
6. 研究方法 -----------------------------------------------------------13
7. 研究結果 -----------------------------------------------------------21
8. 討論 -----------------------------------------------------------30
9. 參考文獻 -----------------------------------------------------------35
10. 附表與附圖 -----------------------------------------------------------42

1.Alexander SPH, Curtis AR, Hill SJ and Kendall DA. A1 adenosine receptor inhibition of cyclic AMP formation and radioligand binding in the guinea-pig cerebral cortex. Br J Pharmacol 113: 1501-1507; 1994a.
2.Alexander SPH, Losinski A, Kendall DA and Hill SJ. A comparison of A2 adenosine receptor stimulated cyclic AMP generation in cerebral cortex and relaxation of precontracted aorta. Br J Pharmacol 111: 185-190; 1994b)
3.Andersson, P. and Bergstrand, H. Antigen-induced bronchial anaphylaxis in actively sensitized guinea-pig: Effect of long-term treatment with sodium cromoglycat and aminophylline. Br J Pharmac 74: 601-609; 1981.
4.Barnes PJ. Is Asthma a Nervous Disease ? Chest 107 (3): 119s-125s; 1995.
5.Barnes PJ. Modulation of neurotransmission in airway. Physiol Rev 72: 699-729; 1992a.
6.Barnes PJ. Neural control of airways function: new perspectives. Molec Aspects Med 11: 351-423; 1990.
7.Barnes PJ. Neural control of human airways in health and disease. Am Rev Respir Dis 134: 1289-1314; 1986.
8.Barnes PJ. New drugs for asthma. Eur Respir J 5: 1126-1136; 1992.
9.Barnes PJ, Baraniuk J and Belvisi MG. Neuropeptides in the respiratory tract. Am Rev Respir Dis 144 : 1187-1198; 1991.
10.Barnes PJ, Baraniuk J, Belvisi MG. Neuropeptides in the respiratory tract. Am Rev Respir Dis 144: 1187-1198, 1391-1399; 1991.
11.Barnes PJ, Belvisi MG and Rogers DF. Modulation of neurogenic inflammation: novel approaches to inflammatory diseases. Trends Pharmacol Sci 11: 185-189; 1990.
12.Bevan S. and Yeats JC. Protons activate a sustained inward current in a sub-population of rat isolated trigeminal ganglion (TG) neurones. Physiol 433: 145-161; 1991.
13.Buck SH. and Burks TF. The neuropharmacology of capsaicin: review of some recent observations. Pharmacol Rev 38: 179-226; 1986.
14.Chand N, Nolan K, Pillar J, Lomask M, Diamants W. and Sofia RD. Aeroallergen-induced dyspnea in Freely moving guinea pigs : quantitative measurement by bias flow ventilated whole body plethysmography. Allergy 48: 230-235; 1993.
15.Chen IJ, Yang JM, Yeh JL, Wu BN, Lo YC. and Chen SJ. Hypotensive and antinociceptive effects of eitherlinked and relatively non-pungent analogues of N-nonanoyl vanillylamide. Eur J Med Chem 27: 187-192; 1992.
16.Colasurdo GN, Loader JE, Graves JP. and Larsen GL. Modulation of acetylcholine release in rabbit airway in vitro. Am J Physiol 268 (Lung Cell. Mol. Physiol. 12): L432-437; 1995.
17.Collier JG. and Fuller RW. Capsaicin inhalation in man and the effects of sodium cromoglycate. Br J Pharmacol 81: 113-117; 1984.
18.Collis MG, Hourani SMO. Adenosine receptor subtypes. Trends Pharmacol Sci 14: 360-366; 1993.
19.Delpierre S, Grimaud CH, Jammes Y. and Mei N. Changes in activity of vagal bronchopulmonary C fibres by chemical and physical stimuli in the cat. J physiol (Loden) 316: 61-74; 1981.
20.Ellis JL. and Undem BJ. Pharmacology of non-adrenergic, non-cholinergic nerve in airway smooth muscle. Pulmon Pharmacol 7: 205-24; 1994.
21.Gamse R, Holzer P. and Lembeck F. Decrease of substance P in primary afferent neurones and impairment of neurogenic plasma extravasation by capsaicin. Br J Pharmacol 68: 207-13; 1980.
22.Ishida K, Kelly LJ, Thomson RJ, Beattie LL. and Schellenberg RR. Repeated antigen challenge induces airway hyperresponsiveness with tissue eosinophilia in guinea pigs. J Appl Physiol 67: 1133-9; 1989.
23.Karlsson JA, Sain`Ambrogio G. and Widdicombe J. Afferent neural pathways in cough and reflex bronchoconstriction. J Appl Physiol 65: 1007-23; 1998.
24.Knox, Aj. And Tattersfield, AE. Airway smooth muscle. In: Szekeres, L., Papp, J.G. (Eds.), Handbook of Experimental Pharmacolgy, Vol. 111. Springger-Verlag, Berlin, pp. 405-443; 1994.
25.Kotlikoff, MI. Potassium channels in airway smooth muscle: a tonic airway smooth muscle: initiation of phasic electrical activity. Am J Physiol 228: 633-636; 1975.
26.Lilly CM, Bai TR, Shore SA, Hall AE. and Drazen JM. Neuropeptide content of lungs from asthmatic and nonasthmatic patients. Am J Respir Crit Care Med 151: 548-553; 1995.
27.Lo YC, Huang W. and Chen IJ. Glyceryl Nonivamide, a Nonpungent Analogue of Capsaicin, Enhances Substance P Release and Renal Function in Rats. Pharmacology 54: 127-134; 1997.
28.Lo YC, Wu JR, Wu SN. and Chen IJ. Glyceryl Nonivamide : A Capsaicin Derivative with Cardiac Calcitonin Gene-Related Peptide Releasing, K+ Channel Opening and Vasorelaxation Properties. J Pharmacol Exp Thera 281: 253-260; 1997.
29.Lundberg JM, Brodin E. and Saria A. Effects and distribution of vagal capsaicin-sensitive substance P neurons with special reference to the trachea and lungs. Acta Physiol Scand 119: 243-52; 1983.
30.Lundberg JM, Martling CR and Saria A. Substance P and capsaicin-induced contraction of human bronchi. Acta Physiol Scand 119: 49-53; 1983.
31.Maggi CA. Tachykinins and calcitonin gene-related peptide (CGRP) as co-transmitters released from peripheral endings of sensory nerves. Prog Neurobiol 48: 1-98; 1995.
32.Manzini S. Bronchodilatation by tachykinins and capsaicin in the mouse main bronchus. Br J Pharmacol 105: 968-972; 1992.
33.Maria G, Belvisi C, David S, Magdi Y. and Barnes PJ. Nitric oxide is the endogenous neurotransmitter of bronchodilator nerve in humans. Eur J Pharmacol 210: 221-222; 1992.
34.Mariem EL Sayah, Valdir Cechinel Filho, Rosendo A. Yunes and Joao B. Calixto. Action of the Extract of Drymis winteri on Contraction Induced by Inflammatiory Mediators, Compound 48/80 and Ovalbumin of the Guinea-Pig Trachea In Vitro. Gen Pharmac 28(5): 699-704; 1997.
35.Matsuse T, Thompson RJ, Chen XR, Salari H. and Schellenberg RR. Capsaicin inhibits airway hyperresponsiveness but not lipoxygenase activity or eosinophilia after repeated aerosolized antigen in guinea pigs. Am Rev Respir Dis 144: 368-72; 1991.
36.Myer AC. and Undem BJ. Electrophysiological effects of tachykinins and capsaicin on guinea-pig bronchial parasympathetic ganglion neurones. J Phsiol (Lond.) 470: 665-679; 1993.
37.O`Neill TP. Mechanism of capsaicin action : recent learnings. Respir Med 85: 35-41; 1991.
38.Ollerenshaw SL, Javis D, Sullivan CE. and Woolcock AJ. Substance P immunoreactive nerves in airways from asthmatics and non-asthmatics. Eur Respir Med 85: 35-41; 1991.
39.Pretolani M, Lefort J, Dumarey C. and Vargaftig B.B. Role of lipoxygenaes metabolites for the hyper-responsiveness to platelet-activating factor of lungs from actively sensitized guinea-pig. J Pharmac Exp Ther 248: 353-359; 1989.
40.Schelemper V. and Calixto JB. Mechanisms involved in the relaxant response of bradykinin in epithelium intact strips of the guinea-pig trachea. Eur J Pharmac 282: 177-184; 1995.
41.Sonea IM, Bowker RM, Robinson NE. and Broadstone RV. Substance P and calcitonin gene-related peptide-like immunoreactive nerve fibers in lung from adult equides. Am J Vet Res 55: 1066-1074; 1994.
42.Spina D. Airway sensory nerve: a burning issue in asthma? Thorax 51(3): 335-337; 1996.
43.Spina D, McKenniff MG, Coyle AJ, Seeds EAM, Perretti F, Tramontana M, et al. Effect of capsaicin on PAF-induced bronchial hyperresponsiveness and pulmonary cell accumulation in the rabbit. Br J Pharmacol 102: 1268-74; 1991.
44.Stead RH, Perdue MH. and Blennerhassett MG. The innervation of mast cell. In: The Neuroendocrine-Immune Network, edited by S. Freier. Boca Raton, FL: CRC, p.19-37; 1990.
45.Stretton CD, Miura M, Belvisi MG. and Barnes PJ. Calcium-activated potassium channels mediate prejunctional inhibition of peripheral sensory nerve. Proc Natl Acad Sci USA 89: 1325-1329; 1992.
46.Szarek JL, Stewart NL, Spurlock B. and Schneider C. Sensory nerve- and neuropeptide-mediated relaxation responses in airways of Sprague-Dawley rats. J Appli Physiol 78(5): 1679-87; 1995.
47.Tater M. and Widdicombe JG. Lung C-fibrereceptor activation and defensive reflexes in anaesthetized cats. J Physiol (Lond) 402: 411-420; 1988.
48.Therieut E, Otsuka M. and Jessel T. Capsaicin evoked release of substance P from primary sensory neurones. Brain Res 170: 209-213; 1979.
49.Yang JM, Tang CS, Yu KL, Yang YC, Chang WC. and Chen IJ. Sodium Nonivamide Acetate: A Non-Pungently Antinociceptive Capsaicin Derivative With Unusual Anti-Inflammatory Properties. Gen Pharmac 27(1): pp.141-150; 1996.
50.Yu M, Robinson NE, Wang Z. and Derksen FJ. Independent modulation of horse airway smooth muscle by epithelium and prostanoids. Respir Physiol 93: 279-288; 1993.
51.Yu M, Wang Z, Robinson NE. and Leblanc PH. Inhibitory nerve distribution and mediation of NANC relaxation by nitric oxide in horse airways. J Appl Physiol 76: 339-334; 1994.
52.Zhu FX, Zhang XY, Olszewski MA. and Robinson NE. Mechanism of capsaicin-induced relaxation in equine tracheal smooth muscle. Am J Physiol 273(5 Pt 1): L997-1001; 1997.