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標題: 肥大細胞釋出液對缺血腦組織保護機轉之探討
The mechanism(s) underlying mast cell-derived conditioned medium(MaCM)-mediated brain protection against cerebral ischemia
作者: 吳姿璉
Wu, Tze-Line
關鍵字: cerebral ischemia;腦缺血;GOSD;mast cell;MaCM;histamine;BBB;neutrophil;GOSD;肥大細胞;肥大細胞釋出液(MaCM);組織胺;血腦障壁;嗜中性白血球
出版社: 生命科學系所
引用: 一、 中文部份 行政院衛生署, 2007。死因統計結果 二、英文部份: Abbott, N.J., 2002. Astrocyte‐endothelial interactions and blood‐brain barrier permeability. J Anat. 200, 629‐38. Adachi, N., 2005. Cerebral ischemia and brain histamine. Brain Res Brain Res Rev. 50,275‐86. Adams, H.P., Jr., 2002. Emergent use of anticoagulation for treatment of patients with ischemic stroke. Stroke. 33, 856‐61. Azizkhan, R.G., Azizkhan, J.C., Zetter, B.R., Folkman, J., 1980. Mast cell heparin stimulates migration of capillary endothelial cells in vitro. J Exp Med. 152, 931‐44. Azuma, Y., Shinohara, M., Wang, P.L., Hidaka, A., Ohura, K., 2001. Histamine inhibits chemotaxis, phagocytosis, superoxide anion production, and the production of TNFalpha and IL‐12 by macrophages via H2‐receptors. Int Immunopharmacol. 1,1867‐75. Bankl, H.C., Radaszkiewicz, T., Klappacher, G.W., Glogar, D., Sperr, W.R., Grossschmidt, K., Bankl, H., Lechner, K., Valent, P., 1995. Increase and redistribution of cardiac mast cells in auricular thrombosis. Possible role of kit ligand. Circulation. 91, 275‐83. Bischoff, S.C., Sellge, G., Lorentz, A., Sebald, W., Raab, R., Manns, M.P., 1999. IL‐4 enhances proliferation and mediator release in mature human mast cells. Proc Natl Acad Sci U S A. 96, 8080‐5. Blair, R.J., Meng, H., Marchese, M.J., Ren, S., Schwartz, L.B., Tonnesen, M.G., Gruber, B.L., 1997. Human mast cells stimulate vascular tube formation. Tryptase is a novel, potent angiogenic factor. J Clin Invest. 99, 2691‐700. Bradding, P., Feather, I.H., Wilson, S., Bardin, P.G., Heusser, C.H., Holgate, S.T., Howarth, P.H., 1993. Immunolocalization of cytokines in the nasal mucosa of normal and perennial rhinitic subjects. The mast cell as a source of IL‐4, IL‐5, and IL‐6 in human allergic mucosal inflammation. J Immunol. 151, 3853‐65. Bradding, P., Okayama, Y., Howarth, P.H., Church, M.K., Holgate, S.T., 1995.Heterogeneity of human mast cells based on cytokine content. J Immunol. 155,297‐307. Bury, T.B., Radermecker, M.F., 1989. Depression of polymorphonuclear chemotaxis and T‐lymphocyte proliferation following histamine inhalation in man. Eur Respir J. 2,52 828‐33. Cannon, K.E., Leurs, R., Hough, L.B., 2007. Activation of peripheral and spinal histamine H3 receptors inhibits formalin‐induced inflammation and nociception,respectively. Pharmacol Biochem Behav. 88, 122‐9. Chu, L.F., Wang, W.T., Ghanta, V.K., Lin, C.H., Chiang, Y.Y., Hsueh, C.M., 2008. Ischemic brain cell‐derived conditioned medium protects astrocytes against ischemia through GDNF/ERK/NF‐kB signaling pathway. Brain Res. 1239, 24‐35. Conti, P., DiGioacchino, M., 2001. MCP‐1 and RANTES are mediators of acute and chronic inflammation. Allergy Asthma Proc. 22, 133‐7. Crivellato, E., Ribatti, D., 2005. Involvement of mast cells in angiogenesis and chronic inflammation. Curr Drug Targets Inflamm Allergy. 4, 9‐11. Danilewicz, M., Wagrowska‐Danilewicz, M., 2004. Immunohistochemical analysis of the interstitial mast cells in acute rejection of human renal allografts. Med Sci Monit. 10, BR151‐6. Deli, M.A., 2009. Potential use of tight junction modulators to reversibly open membranous barriers and improve drug delivery. Biochim Biophys Acta. 1788,892‐910. Dirnagl, U., Iadecola, C., Moskowitz, M.A., 1999. Pathobiology of ischaemic stroke: an integrated view. Trends Neurosci. 22, 391‐7. Dvorak, A.M., Dvorak, H.F., Galli, S.J., 1983. Ultrastructural criteria for identification of mast cells and basophils in humans, guinea pigs, and mice. Am Rev Respir Dis. 128, S49‐52. Dvorak, A.M., Kissell, S., 1991. Granule changes of human skin mast cells characteristic of piecemeal degranulation and associated with recovery during wound healing in situ. J Leukoc Biol. 49, 197‐210. Farmer, L., 1940. Histamine in Anaphylaxis and Allergy. Bull N Y Acad Med. 16, 618‐30. Fox, S., Shephard, T.J., McCain, N., 1999. Neurologic mechanisms in psychoneuroimmunology. J Neurosci Nurs. 31, 87‐96. Frangogiannis, N.G., Lindsey, M.L., Michael, L.H., Youker, K.A., Bressler, R.B., Mendoza, L.H., Spengler, R.N., Smith, C.W., Entman, M.L., 1998. Resident cardiac mast cells degranulate and release preformed TNF‐alpha, initiating the cytokine cascade in experimental canine myocardial ischemia/reperfusion. Circulation. 98, 699‐710. Fraser, R.A., Simpson, J.G., 1983. Role of mast cells in experimental tumour angiogenesis. Ciba Found Symp. 100, 120‐31. Galli, S.J., 1990. New insights into "the riddle of the mast cells": microenvironmental regulation of mast cell development and phenotypic heterogeneity. Lab Invest. 62, 5‐33.53 Galli, S.J., Tsai, M., 2008. Mast cells: versatile regulators of inflammation, tissue remodeling, host defense and homeostasis. J Dermatol Sci. 49, 7‐19. Garbarg, M., Barbin, G., Bischoff, S., Pollard, H., Schwartz, J.C., 1976. Dual localization of histamine in an ascending neuronal pathway and in non‐neuronal cells evidenced by lesions in the lateral hypothalamic area. Brain Res. 106, 333‐48. Goldschmidt, R.C., Hough, L.B., Glick, S.D., Padawer, J., 1984. Mast cells in rat thalamus: nuclear localization, sex difference and left‐right asymmetry. Brain Res. 323,209‐17. Goldschmidt, R.C., Hough, L.B., Glick, S.D., 1985. Rat brain mast cells: contribution to brain histamine levels. J Neurochem. 44, 1943‐7. Hebda, P.A., Collins, M.A., Tharp, M.D., 1993. Mast cell and myofibroblast in wound healing. Dermatol Clin. 11, 685‐96. Hechtman, D.H., Cybulsky, M.I., Fuchs, H.J., Baker, J.B., Gimbrone, M.A., Jr., 1991.Intravascular IL‐8. Inhibitor of polymorphonuclear leukocyte accumulation at sites of acute inflammation. J Immunol. 147, 883‐92. Heldmann, U., Thored, P., Claasen, J.H., Arvidsson, A., Kokaia, Z., Lindvall, O., 2005. TNF‐alpha antibody infusion impairs survival of stroke‐generated neuroblasts in adult rat brain. Exp Neurol. 196, 204‐8. Hiraga, N., Adachi, N., Liu, K., Nagaro, T., Arai, T., 2007. Suppression of inflammatory cell recruitment by histamine receptor stimulation in ischemic rat brains. Eur J Pharmacol. 557, 236‐44. Hu, W., Xu, L., Pan, J., Zheng, X., Chen, Z., 2004. Effect of cerebral ischemia on brain mast cells in rats. Brain Res. 1019, 275‐80. Hu, W., Shen, Y., Fu, Q., Dai, H., Tu, H., Wei, E., Luo, J., Chen, Z., 2005. Effect of oxygen‐glucose deprivation on degranulation and histamine release of mast cells. Cell Tissue Res. 322, 437‐41. Iadecola, C., Alexander, M., 2001. Cerebral ischemia and inflammation. Curr Opin Neurol. 14, 89‐94. Ioffreda, M.D., Whitaker, D., Murphy, G.F., 1993. Mast cell degranulation upregulates alpha 6 integrins on epidermal Langerhans cells. J Invest Dermatol. 101, 150‐4. Kadam, S.D., Mulholland, J.D., Smith, D.R., Johnston, M.V., Comi, A.M., 2009. Chronic brain injury and behavioral impairments in a mouse model of term neonatal strokes. Behav Brain Res. 197, 77‐83. Kamei, C., 2001. Involvement of central histamine in amygdaloid kindled seizures in rats.Behav Brain Res. 124, 243‐50. Katsanos, G.S., Anogeianaki, A., Orso, C., Tete, S., Salini, V., Antinolfi, P.L., Sabatino, G., 2008. Mast cells and chemokines. J Biol Regul Homeost Agents. 22, 145‐51. 54 Kovacic, J.C., Gupta, R., Lee, A.C., Ma, M., Fang, F., Tolbert, C.N., Walts, A.D., Beltran, L.E., San, H., Chen, G., St Hilaire, C., Boehm, M., Stat3‐dependent acute Rantes production in vascular smooth muscle cells modulates inflammation following arterial injury in mice. J Clin Invest. 120, 303‐14. Lamalice, L., Le Boeuf, F., Huot, J., 2007. Endothelial cell migration during angiogenesis.Circ Res. 100, 782‐94. Lee, G.A., Lin, C.H., Jiang, H.H., Chao, H.J., Wu, C.L., Hsueh, C.M., 2004.Microglia‐derived glial cell line‐derived neurotrophic factor could protectSprague‐Dawley rat astrocyte from in vitro ischemia‐induced damage. Neurosci Lett. 356, 111‐4. Levi‐Schaffer, F., Pe''er, J., 2001. Mast cells and angiogenesis. Clin Exp Allergy. 31, 521‐4. Lin, C.H., Cheng, F.C., Lu, Y.Z., Chu, L.F., Wang, C.H., Hsueh, C.M., 2006. Protection of ischemic brain cells is dependent on astrocyte‐derived growth factors and their receptors. Exp Neurol. 201, 225‐33. Martres, M.P., Baudry, M., Schwartz, J.C., 1975. Histamine synthesis in the developing rat brain: evidence for a multiple compartmentation. Brain Res. 83, 261‐75. Meng, H., Tonnesen, M.G., Marchese, M.J., Clark, R.A., Bahou, W.F., Gruber, B.L., 1995.Mast cells are potent regulators of endothelial cell adhesion molecule ICAM‐1 and VCAM‐1 expression. J Cell Physiol. 165, 40‐53. Miyamoto, K., Iwase, M., Nyui, M., Arata, S., Sakai, Y., Gabazza, E.C., Kimura, H., Homma, I., 2006. Histamine type 1 receptor deficiency reduces airway inflammation in a murine asthma model. Int Arch Allergy Immunol. 140,215‐22. Moller, A., Lippert, U., Lessmann, D., Kolde, G., Hamann, K., Welker, P., Schadendorf, D., Rosenbach, T., Luger, T., Czarnetzki, B.M., 1993. Human mast cells produce IL‐8. J Immunol. 151, 3261‐6. Nienartowicz, A., Sobaniec‐Lotowska, M.E., Jarocka‐Cyrta, E., Lemancewicz, D., 2006. Mast cells in neoangiogenesis. Med Sci Monit. 12, RA53‐6. Norrby, K., 2002. Mast cells and angiogenesis. APMIS. 110, 355‐71. Otsuka, R., Adachi, N., Hamami, G., Liu, K., Yorozuya, T., Arai, T., 2003. Blockade of central histaminergic H2 receptors facilitates catecholaminergic metabolism and aggravates ischemic brain damage in the rat telencephalon. Brain Res. 974,117‐26. Pang, X., Letourneau, R., Rozniecki, J.J., Wang, L., Theoharides, T.C., 1996. Definitive characterization of rat hypothalamic mast cells. Neuroscience. 73, 889‐902. Reed, J.A., Albino, A.P., McNutt, N.S., 1995. Human cutaneous mast cells express basic fibroblast growth factor. Lab Invest. 72, 215‐22.55 Ribatti, D., Roncali, L., Nico, B., Bertossi, M., 1987. Effects of exogenous heparin on the vasculogenesis of the chorioallantoic membrane. Acta Anat (Basel). 130, 257‐63. Rosenberg, G.A., Estrada, E.Y., Dencoff, J.E., 1998. Matrix metalloproteinases and TIMPs are associated with blood‐brain barrier opening after reperfusion in rat brain. Stroke. 29, 2189‐95. Schoch, H.J., Fischer, S., Marti, H.H., 2002. Hypoxia‐induced vascular endothelial growth factor expression causes vascular leakage in the brain. Brain. 125,2549‐57. Sorbo, J., Jakobsson, A., Norrby, K., 1994. Mast‐cell histamine is angiogenic through receptors for histamine1 and histamine2. Int J Exp Pathol. 75, 43‐50. Takahashi, H.K., Yoshida, A., Iwagaki, H., Yoshino, T., Itoh, H., Morichika, T., Yokoyama, M., Akagi, T., Tanaka, N., Mori, S., Nishibori, M., 2002. Histamine regulation of interleukin‐18‐initiating cytokine cascade is associated with down‐regulation of intercellular adhesion molecule‐1 expression in human peripheral blood mononuclear cells. J Pharmacol Exp Ther. 300, 227‐35. Thompson, H.L., Thomas, L., Metcalfe, D.D., 1993. Murine mast cells attach to and migrate on laminin‐, fibronectin‐, and matrigel‐coated surfaces in response to Fc epsilon RI‐mediated signals. Clin Exp Allergy. 23, 270‐5. Tomita, M., Matsuzaki, Y., Onitsuka, T., 2000. Effect of mast cells on tumor angiogenesis in lung cancer. Ann Thorac Surg. 69, 1686‐90. Tosaki, A., Szerdahelyi, P., Joo, F., 1994. Treatment with ranitidine of ischemic brain edema. Eur J Pharmacol. 264, 455‐8. Tuttolomondo, A., Di Sciacca, R., Di Raimondo, D., Renda, C., Pinto, A., Licata, G., 2009. Inflammation as a therapeutic target in acute ischemic stroke treatment. Curr Top Med Chem. 9, 1240‐60. Wahl, M., Unterberg, A., Baethmann, A., Schilling, L., 1988. Mediators of blood‐brain barrier dysfunction and formation of vasogenic brain edema. J Cereb Blood Flow Metab. 8, 621‐34. Wolburg, H., Lippoldt, A., 2002. Tight junctions of the blood‐brain barrier: development, composition and regulation. Vascul Pharmacol. 38, 323‐37. Zauberman, H., Michaelson, I.C., Bergmann, F., Maurice, D.M., 1969. Stimulation of neovascularization of the cornea by biogenic amines. Exp Eye Res. 8, 77‐88
肥大細胞(mast cell)一般都被認為是-促發炎細胞,可釋放組織胺
(histamine)造成不同組織包括缺血腦組織的發炎反應。但近年來透過對不同組織胺受體的研究,發現組織胺可經由活化H2 受體保護缺血腦組織,使肥大細胞的角色另起爭議。本論文主要在評估肥大細胞釋出液(mast cell-derived condition medium or MaCM),對缺血腦組織是否具保護性及其作用機轉,以進一步釐清肥大細胞在缺血腦組織中所扮演的角色及調控機轉。實驗中,利用結紮大白鼠腦部血管的體內缺血/再灌流模式,及細胞缺糖、缺氧、缺血清(glucose-oxygen- and serum-deprivation or GOSD)的體外缺血模式,做為研究平台,針對肥大細胞在缺血腦組織中及體外GOSD環境壓力下,其存活力、數量及釋放組織胺的能力等變化做分析。接著更將經GOSD處理後所產生之MaCM 或組織胺,送入缺血腦組織皮質區,評估兩者對缺血鼠之腦組織及運動功能的受損,血腦障壁(blood brain barrier or BBB)的被破壞,及腦缺血區occludin 及血管生長因子(VEGF)的蛋白表現等,是否有保護功能。而MaCM 或組織胺,對經GOSD 處理後之腦細胞(神經細胞、星芒狀細胞、微膠細胞)及內皮細胞的存活力或移動力之影響;或對缺血鼠脾臟中嗜中性白血球的移動力及氧化活性的影響,亦在評估之列。研究結果顯示,在GOSD 的壓力下,肥大細胞的存活明顯降低,但其釋放組織胺的能力顯著提高。大鼠右大腦半球缺血皮質區,肥大細胞的數目及去顆粒化現象和左大腦半球之皮質區相比並無明顯差異,但在右大腦半球的視丘區,肥大細胞去顆粒化及釋放組織胺的現象明顯提升。MaCM 對缺血腦組織明顯具保護性,此保護作用除和H3 受體之活化有關外,亦可能經由抑制VEGF 和提升occludin 的蛋白表現,來降低缺血區BBB 的通透性,及抑制嗜中性白血球的移動力或氧化活性所致。MaCM 對於GOSD 所導致的腦細胞受損,及缺血所導致的運動功能受損情形,則不具保護性。整體而言,肥大細胞在面對缺血壓力時,可經由穩固BBB 結構、抑制促發炎細胞(嗜中性白血球)的氧化活性及移動力等特質,來降低缺血腦組織的受損程度。肥大細胞的活化(去顆粒化)明顯對缺血組織具保護功能,部分是經由釋放組織胺活化H3 受體所致。此外,MaCM 在缺血性腦中風的治療上亦具療效,其保護功能除部分仰賴組織胺外,明顯亦依賴其他分子的參與,如能進一步釐清MaCM 中具保護功能的其他成份,對未來抗中風藥物的篩選或創新研發上,均具參考及應用價值。

Mast cell has been considered as a pro-inflammatory cell and through the release of histamine it actively involved in inflammation of many tissues including ischemic brain. Nevertheless, an anti-inflammatory activity of histamine receptor (H2 subtype)has recently been suggested to protect brain from ischemic insult may also reflect an anti-inflammatory role for mast cells in cerebral ischemia. The primary goal of the thesis was to assess the mechanism(s) underlying mast cell-derived conditioned medium (MaCM)-mediated brain protection against cerebral ischemic. Through the study, the regulatory role (pro- or anti-inflammatory) of mast cell in cerebral ischemia can be further delineated. In the study, a reversible focal ischemia was first developed in Spraque‐Dawley (S.D.) rats by subjecting animals to 90 min of bilateral common carotid artery occlusion (CCAO) plus unilateral middle cerebral artery occlusion (MCAO) followed by reperfusion for 24h. In the meantime, primary neurons, astrocytes and microglia were isolated and exposed to glucose‐ oxygen‐ and serum‐deprivation (GOSD)condition to mimic cerebral ischemia in vitro. Both experimental platforms were applied to analyze the viability, number and histamine releasing capability of mast cells under either ischemic or GOSD condition. The protective effect of MaCM or histamine on ischemic brain, motor activity, blood brain barrier (BBB) or the expression of occludin and vascular endothelial growth factor (VEGF) were further examine by injecting MaCM or histamine directly into the core area (cortex) of ischemic brain.
The effects of MaCM or histamine on: survival of GOSD brain cells (neuron, astrocyte and microglia) and endothelial cells, motility of GOSD endothelial cells and neutrophils, and the release of superoxide from neutrophils were also investigated in vitro under GOSD condition. Our results showed that GOSD inhibited viability of but increased histamine release from mast cells. The number and degranulation of mast cells at cortical region of ipsilateral hemisphere remained similar to that at same region of contralateral hemisphere. Degranulation and the release of histamine from mast cells however, were significantly increased at thalamus of ipsilateral hemisphere. MaCM collected at 6 h after GOSD, significantly reduced brain infarct volume caused by ischemia/reperfusion, in a histamine receptor (H3-subtype)-dependent manner. MaCM-mediated brain protection may act through the inhibition of VEGF and stimulation of occludin expression, reduction of BBB permeability, and inhibition of neutrophil motility and release of superoxide. MaCM however, did not protect brain cells from GOSD insult in vitro, nor restored motor activity of ischemic rats in vivo.In summary, in response to ischemic stress, the activation of mast cells appeared to be brain protective by releasing substances such as histamine and/or other unidentified factors to stabilize BBB and inhibit brain entrance and oxidative activity of neutrophils that eventually led to significant reduction of brain infarction. MaCM appeared to have therapeutic value in the control of cerebral ischemia. Identification of other protective substances within MaCM may provide additional choices for future design of new anti-ischemic drugs.
其他識別: U0005-0802201011323300
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