本篇論文中我們利用Long Evans品系之大鼠研究6-hydroxydopamine (6-OHDA)引發膀胱以及前列腺發炎，並探討其中之機轉。我們利用兩種方法來評估發炎反應所引起之血漿滲漏: 第一，我們利用India ink標示滲漏血管來統計膀胱壁滲漏血管密度；第二，我們利用Evans blue dye滯留在組織中之含量來表示膀胱壁以及前列腺血漿滲漏程度。在膀胱整體封裝標本，觀察被India ink標示滲漏血管密度顯示，對照組、6-OHDA 5mg/kg、6-OHDA 10mg/kg各會造成之滲漏血管密度為5.65±1.72 % (N=6), 22.63±3.12 % (N=6), 35.02±2.25 % (N=6)。利用Evans blue dye滯留在組織中之含量來表示膀胱壁以及前列腺血漿滲漏程度，6-OHDA 5mg/kg會造成膀胱壁中Evans blue dye 80.53±60.74 ng/mg滯留，而前列腺48.81±2.83 ng/mg滯留。 6-OHDA 10mg/kg會造成膀胱壁中Evans blue dye 157.73±4.45 ng/mg滯留，而前列腺65.52±4.25 ng/mg滯留。對照組相對只有少量Evans blue dye滯留，膀胱壁中Evans blue dye 18.82±3.74 ng/mg，而前列腺 18.50±2.47 ng/mg。實驗結果證明6-OHDA確實會引起膀胱以及前列腺發炎。並且隨著6-OHDA劑量增加，膀胱以及前列腺血漿滲漏程度亦會增加。我們利用自由基清除劑dimethylthiourea幾乎可以完全抑制6-OHDA之發炎反應，然而tachykinin類 (NK1 receptor)接受器拮抗劑L-732,138卻只是中等程度抑制6-OHDA之發炎反應。我們也利用掃描式電子顯微鏡觀察膀胱壁橫切面中的血管腔以及前列腺腺體內腔，掃描式電子顯微鏡觀察中發現:6-OHDA會引起膀胱壁橫切面中的血管腔內皮細胞間隙(gap junction)打開並且前列腺腺體內腔分泌顆粒增多。本篇研究結論: 6-OHDA確實會造成膀胱以及前列腺血漿滲漏。其中6-OHDA所產生之自由基是引發血漿滲漏的關鍵因子，另外tachykinin在其中扮演次要的角色。

The mechanisms underlying the 6-hydroxydopamine (6-OHDA)-induced inflammatory response in the urinary bladder and prostate in anaesthetized male rats of Long- Evan strain were investigated. The magnitude of inflammatory responses were evaluated by morphometric analysis of the area density of India ink-labeled blood vessels in urinary bladder whole mounts and spectrophotometric analysis of Evans blue dye contents in urinary bladder and prostate. Moreover, scanning electron microscopy was employed to observe the venular endothelium in the urinary bladder wall and glandular epithelium in the prostate gland. Fifteen minutes after local application of 6-OHDA to the urinary bladder, 6-OHDA induced an increase of plasma leakage in a dose-dependent manner. It was revealed that area densities of India ink-labeled blood vessels in the rat urinary bladder whole mount were 5.65±1.72 % (N=6), 22.63±3.12 % (N=6), and 35.02±2.25 % (N=6) respectively, following a local injection of vehicle, 5 mg/kg 6-OHDA, and 10 mg/kg 6-OHDA. Using Evans blue dye as a tracer for spectrophotometric analysis, the results were similar. The Evans blue dye content was 80.53±60.74 ng/mg in the urinary bladder and 48.81±2.83 ng/mg in the prostate following injection of 5 mg/kg 6-OHDA (N=6). The Evans blue dye content was 157.73±4.45 ng/mg in the bladder and 65.52±4.25 ng/mg in the prostate following injection of 10 mg/kg 6-OHDA (N=6). Evans blue dye contents in the vehicle group (N=6) were much lower, 18.82±3.74 ng/mg in the urinary bladder and 18.50±2.47 ng/mg in the prostate, which were significantly smaller than the 6-OHDA treated group. Interestingly, the inflammatory responses were completely abolished by pretreating alone with dimethylthiourea (DMTU), a hydroxyl radical scavenger, and were moderately attenuated by pretreatment with L-732,138, a NK1 receptor antagonist. Under scanning electron microscope observation, 6-OHDA caused endothelial gaps formation in the venules of urinary bladder wall and triggered the release of secretory granules in the prostate gland cells. We concluded that 6-OHDA could induce inflammation in the urinary bladder and prostate gland involving free radical and tachykinin mechanisms.

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ABSTRACT--------------------------------------------------------6
INTRODUCTION-------------------------------------------------8
MATERIALS AND METHODS--------------------------------12
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Apodaca, G. 2004. The uroepithelium: not just a passive barrier. Traffic 5: 117-128.

Baluk, P., Hirata, A., Thurston, G., Fujiwara, T., Neal, C. R., Michel, C. C., McDonald, D. M. 1997. Endothelial gaps: time course of formation and closure in inflamed venules of rats. Am. J. Physiol. 272 (Lung Cell Mol Physiol 16):L155-L170.

Bharali, L. A. M., Lisney, S. J. W. 1992. The relationship between unmyelinated afferent type and neurogenic plasma extravasation in normal and reinnervated skin. Neuroscience. 47: 703-712.

Bradley, J. R., Thiru, S., Pober, J. S. 1995. Hydrogen peroxide-induced endothelial retraction is accompanied by a loss of the normal spatial organization of endothelial cell adhesion molecules. Am. J. Pathol. 147: 627-641.

Candenas, L., Lecci, A., Pinto, F. M., Patak, E., Maggi, C. A., Pennefather, J. N. 2005. Tachykinins and tachykinin receptors: effect in the genitourinary tract. Life Sci. 76(8):835-62.

Cambridge, H., Ajuebor, M. N., Brain, S. D. 1996. Investigation of 6-hydroxydopamine-induced plasma extravasation in rat skin. Eur. J. Pharmacol. 301:151-157.

Coderee, T. J., Basbaum, A. I., and Levine, J. D. 1989. Neural control of vascular permeability: interactions between primary afferents, mass cells, and sympathetic efferents. J. Neurophysiol. 62: 48-58.

Donnerer, J., Amann, R., Lembeck, F. 1991. Neurogenic and non-neurogenic inflammation in the rat paw following chemical sympathectomy. Neurosci. 45: 761-765.

Eglezos, A. S., Guiliani, G. V., Maggi, C. A. 1991. Direct evidence that capsaicin-induced plasma extravasation is mediated through tachykinin NK-receptors. Eur. J. Pharmacol. 209: 277-279.

Evans, T. W., Rogers, D. F., Aursudkij, B., Chung, K. F., Barnes, P. J. 1988. Inflammatory mediators involved in antigen-induced airway microvascular leakage in guinea pigs. Am. Rev. Respir. Dis. 138: 395-399.

Fox, R.B. 1984. Prevention of granulocyte-mediated oxidant lung injury in rats by a hydroxyl radical scavenger, dimethylthiourea. J. Clin. Invest. 74: 1456-1464.

Fox, R.B., Harada R.N., Tate R.M., Repine J.E. 1983. Prevention of thiourea-induced pulmonary edema by hydroxyl radical scanvengers. J. Appl. Physiol. 55: 1456-1459.

Gabella, G., Davis, C. 1998. Distribution of afferent axons in the bladder of rats. J. Neurocytol. 27: 141-155.

Glinka, Y., Gassen, M., Youdim, M. B. 1997. Mechanism of 6-hydroxydopamine neurotoxicity. J. Neural Trans. 50 (Suppl.): 55-66.

Graham, D. G., Tiffany, S. M., Bell, W. R., Jr., Gutknecht, W. F. 1978. Autoxidation versus Covalent Binding of Quinones as the Mechanism of Toxicity of Dopamine, 6-Hydroxydopamine, and Related Compounds toward C1300 Neuroblastoma Cells in Vitro. Mol. Pharmacol. 14: 644-653.

Habler, H. J., Janig, W., and Koltzenburg, M. 1990. Activation of unmyelinated afferent fibers by mechanical stimuli and inflammation of the urinary bladder in the cat. J. Physiol. 425:545-562.

Heikkila, R.E. and Cohen, G. 1973. 6-hydroxydopamine evidence for superoxide radical as an oxidative intermediate. Science. 181:456-457.

Higgins, J. R., Gosling, J. A. 1989. Studies on the structure and intrinsic innervation of the normal human prostate. Prostate. Supplement 2: 5-16.

Holtz, W. A., and O’Malley, K. L. 2003. Parkinsonian Mimetics Induce Aspects of Unfolded Protein Response in Death of Dopaminergic Neurons. J. Biol. Chem. 278, 19367-19377.

Holzer, P. 1988. Local effector functions of capsaicin-sensitive sensory nerve endings: involvement of tachykinins, calcitonin gene-related peptide and other neuropeptides. Neuroscience. 24:739-768.

Hwang, T., Huang, H. T., Tsao, C. F. 1999. Thoracic vagus section distal to the recurrent laryngeal nerve reduces substance P-immunoreactive innervation in the rat bronchial traa. Anat. Embryol. 200:153-160.

Jackson J.H., White C.W., Parker N.B., Ryan J.W., Repine J.E. 1985. Dimethylthiourea consumption reflects H2O2 concentrations and the severity of acute lung injury. J. Appl. Physiol. 59: 1995-1998.

Jen, P. Y., Dixon, J. S. 1995. Development of peptide-containing nerves in the human fetal prostate gland. J. Anat. 187:169-179.

Jonsson, G., Malmfors, T., Sachs, C. 1975. In 6-Hydroxydopamine as a Denervation Tool in Catecholamine Research: Proceedings of a Conference Held in Goteborg, Sweden, July 17-19, 1975. American Elsevier, Goteborg, Sweden.

Kostrzewa, R. M. and Jacobowitz, D. M. 1974. Pharmacological actions of 6-hydroxydopamine. Pharmacol. Rev. 26: 200-287.

Lai, Y. L. 1990. Oxygen radicals in capsaicin-induced bronchoconstriction. J. Appl. Physiol. 68: 568-573.

Lai, C. J., Kou, Y. R. 1998. Stimulation of vagal pulmonary C-fibers by inhaled wood smoke in rats. J. Appl. Physiol. 84: 30-36.

Lee, A. P., Johnson, K. W. 1997. The non-peptide NK-1 receptor antagonist LY303870 inhibits neurogenic dural inflammation in guinea pigs. Life Sci. 60: 1553-1561.

Lei, Y. H., Barnes, P. J., Rogers, D. F. 1995. Mechanisms and modulation of airway plasma exudation after direct inhalation of cigarette smoke. Am. J. Respir. Crit. Care Med. 151: 1752-1762.

Levine J. D., Taiwo, Y. O., Collins, S. D., Tam, J. K. 1986. Noradrenaline hyperalgesia is mediated through interaction with sympathetic postganglionic neurone terminals rather than activation of primary afferent nociceptors. Nature. 323: 158-160.

Levites, Y., Youdim, M. B., Maor, G. and Mandel, Silvia. 2002. Attenuation of 6-OHDA induced nuclear factor Kappa B activation and cell death by tea extracts in neuronal cultures. Biochemical Pharmacol. 63:21-29.

Lin, Y. S., Kou, Y. R. 2000. Acute neurogenic airway plasma exudation and edema induced by inhaled wood smoke in guinea pigs: role of tachykinins and hydroxyl radical. Eur. J. Pharmacol. 394: 139-148.

Lo, Y. K., Huang, H. T. 1997. A novel procedure for mediastinal vaotomy inhibits neurogenic inflammation in rat bronchial tree. J. auton. Nerv. Syst. 67:79-88.

Maggi, C. A., Parlani, M., Astolfi, M., Santicioli, P., Rovero, P., Abelli, L., Somma, V., Giuliana, S., Regoli, D., Patacchini, R. 1988. Neurokinin receptors in the rat lower urinary tract. J. Pharmacol. Exp. Ther. 246:308-315.

Maggi, C. A. 1995. The mammalian tachykinin receptors. Gen. Pharmacol. 26: 911-944.

Malmfors, T. and Sachs, C. 1968. Degeneration of adrenergic nerves produced by 6-hydroxydopamine. Eur. J. Pharmacol. 3:89-92.

McDonald, D.M. 1994. Endothelial gaps and permeability of venules in rat trachea exposed to inflammatory stimuli. Am. J. Physiol. 266:L61-L83.

Nappi, A. J., Vass, E., Prota, G., Memoli, S. 1995. The effects of hydroxyl radical attack on dopa, dopamine, 6-hydroxydopa and 6-hydroxydopamine. Pigment Cell Res. 8: 283-293.

Prystowsky, J. B., Rege, R. V. 1997. Neurogenic inflammation in cholecystitis. Dig. Dis. Sci. 42(7): 1489-1494.

Ryu, E. J., Harding, H. P., Angelastro, J. M., Vitolo, O. V., Ron, D., and Greene, L. A. 2002. Endoplasmic Reticulum Stress and the Unfolded Protein Response in Cellular Models of Parkinson’ s Disease. J. Neurosci. 22: 10690-10698.

Sulakvelidze, I., Baluk, P., and McDonald, D. M. 1994, Plasma extravasation induced in rat trachea by 6-hydroxydopamine is mediated by sensory nerves, not by sympathetic nerves. J. Appl. Physiol. 76: 701-707.

Szolcsanyi, J. 1990. Capsaicin-sensitive chemoceptive B-afferents: A neural system with dual sensory-efferent function. Behav. Brain. Sci. 13:316.

Thurston, G., Baluk, P., Hirata, A., McDonald, D. M. 1996. Permeability-related changes revealed at endothelial cell borders in inflamed venules by lectin binding. Am. J. Physiol. 271(Heart Circ Physiol 40): H2547-H2562.

Wasil, M., Halliwell, B., Grootveld, M., Moorhouse, C. P., Hutchison, D. C. S., Baum, H. 1987. The specificity of thiourea, dimethylthiourea and dimetyl superoxide as scavengers of hydroxyl radicals. Biochem. J. 243: 867-870.