地塞米松对脊髓背角星形胶质细胞P2Y_1受体表达水平及TNF-α分泌影响的初步研究
|
摘要
疼痛对人类的健康危害甚大。脊髓背角对伤害性信息的传递和调制发挥重要作用。星形胶质细胞在中枢神经系统中广泛分布,参与结构和功能维持、免疫反应以及神经疾病的发病等病理生理过程。研究表明,星形胶质细胞与疼痛有关,脊髓胶质细胞的激活是疼痛发生和持续的关键因素,同时与中枢痛觉过敏的产生与维持密切相关。以往研究已发现,疼痛的发生和维持与嘌呤类P2受体有关,嘌呤类P2Y受体参与伤害性信息传递。本实验室曾俊伟等已经证实,脊髓背角星形胶质细胞表达P2Y_1受体,P2Y_1受体激活能引起炎性致痛因子TNF-α的分泌增加。
地塞米松作为临床常用的糖皮质激素,目前研究多集中于它抑制炎性因子分泌而产生镇痛作用,但机制尚不十分清楚。地塞米松能否通过介导P2Y_1受体抑制脊髓背角星形胶质细胞TNF-α的分泌?需要进一步研究。本实验通过纯化培养大鼠脊髓背角星形胶质细胞,观察不同浓度地塞米松对其P2Y_1受体表达的影响,及地塞米松对ADPβs激活后的背角星形胶质细胞P2Y_1受体表达的影响,并通过其对TNF-α分泌的改变推测可能的机制。
方法:培养并纯化大鼠脊髓背角星形胶质细胞,免疫组织化学观察背角星形胶质细胞P2Y_1受体表达水平的变化;ELISA技术检测背角星形胶质细胞TNF-α的分泌。
结果:
第一部分:
1.大鼠脊髓背角星形胶质细胞的培养与纯化,免疫组织化学鉴定细胞类型。结果表明,脊髓背角星形胶质细胞纯度在98%以上。
2.地塞米松呈剂量依赖性抑制脊髓背角星形胶质细胞P2Y_1受体的表达;糖皮质激素受体阻断剂(RU486)特异性阻断糖皮质激素受体后,可拮抗地塞米松对背角星形胶质细胞P2Y_1受体表达的抑制作用。
3.地塞米松呈剂量依赖性抑制脊髓背角星形胶质细胞TNF-α的分泌。RU486特异性阻断糖皮质激素受体后,可拮抗地塞米松对脊髓背角星形胶质细胞TNF-α分泌的抑制作用。
第二部分:
1.ADPβs激活脊髓背角星形胶质细胞,呈剂量依赖性上调P2Y_1受体的表达,在终浓度为100μmol/L时P2Y_1受体表达最多,并促进背角星形胶质细胞TNF-α分泌;P2Y_1受体阻断剂(MRS2179)特异性阻断P2Y_1受体后,可拮抗ADPβs对P2Y_1受体表达及TNF-α分泌的促进作用。
2.ADPβs激活脊髓背角星形胶质细胞,不同浓度地塞米松可呈剂量依赖性的拮抗ADPβs对背角星形胶质细胞P2Y_1受体表达及TNF-α分泌的促进作用。
结论:
1.地塞米松对脊髓背角星形胶质细胞P2Y_1受体表达及TNF-α的分泌均有抑制作用。
2. ADPβs激活背角星形胶质细胞可以使P2Y_1受体表达上调。
3.地塞米松可抑制激活的脊髓背角星形胶质细胞P2Y_1受体的表达及TNF-α的分泌。
综上所述,地塞米松可能通过抑制P2Y_1受体表达和TNF-α的分泌起到提高痛阈的作用;亦可能通过介导P2Y_1受体抑制炎性因子TNF-α的分泌,说明地塞米松在疼痛状态下起到了镇痛作用。
It is known that pain does harm to health. The spinal dorsal horn plays an important role in nociceptive information transmission from the periphery to central nervous system. While neuronal functioning is altered, there is significant evidence showing that exaggerated pain is regulated by the activation of astrocytes and microglia. More and more data suggest that the activation of spinal cord glia is necessary for the development and maintenance of pathological pain and hyperalgesia.Moreover, envidences have revealed that the development and maintenance of pathological pain has something to do with P2 receptor, which take part in nociceptive information transmission.P2Y_1 receptor express at astrocytes which located in spinal dorsal horn, and activation of P2Y_1 recetor leads to the secretion of TNF-α.
Resesrches on dexamethasone have focused on inhibiting inflammatory factor, however mechanism is unknown.Can dexamethasone ihibit the secretion of TNF-αthrough P2Y_1 receptor pathway,and through activating P2Y_1 receptor pathway?This study will use some methods includeing morphology and ELISA to investigate the relationship between dexamethasone and P2Y_1 receptor in cultured rat dorsal horn astrocytes.
Methods: Primary dissociated culture of dorsal spinal cord astrocytes were prepared from Sprague–Dawley rats. The changes of P2Y_1 receptor which expressed in cultured dorsal horn astrocytes were observed by using immunohistochemical staining.And to observe the secretion of TNF-αby means of ELISA.
Results:
Part one:
1. To observe the puration of astrocytes by using immunohistochemical staining,and that of tham was more than 98%.
2. Dexamethasone (0.01~100μmol/L) can dose-dependently decrease the expression of P2Y_1 receptor in cultured rat dorsal horn astrocytes;While after RU486 (1000μmol/L) block the glucocorticoid reccptor, the wok resulted from Dexamethasone become weaker.
3. Dexamethasone (0.01~100μmol/L) can dose-dependently decrease the secretion of TNF-αin cultured rat dorsal horn astrocytes;While after RU486 (100μmol/L) block the glucocorticoid reccptor, the wok resulted from Dexamethasone become weaker.
Part two:
1. ADPβwhich activates cultured dorsal horn atrocytes can dose-dependently increase the expression of P2Y_1 receptor and the secretion of TNF-α; MRS2179(0.1~1000μmol/L), P2Y_1-specific antagonist, can inhibit increasing of the expression of P2Y_1 receptor and the secretion of TNF-α.
2. After ADPβs activated cultured dorsal horn atrocytes, Dexamethasone can dose-dependently the work laeded by ADPβs.
Conclusions:
1. Dexamethasone decrease the expression of P2Y_1 receptor and the secretion of TNF-α.
2. Astrocytes activated by ADPβs can increase the expression of P2Y_1 receptor.
3. Dexamethasone can inhibit the expression of P2Y_1 receptor and the secretion of TNF-αin activated astrocyte.
In summary, these results suggest that Dexamethasone may increase the pain threshold by inhibiting the expression of P2Y_1 receptor and secretion of TNF-αin cultured dorsal horn astrocytes ;We also provide evidence that Dexamethasone can release the pain threshold by inhibiting the secretion of TNF-α.
地塞米松作为临床常用的糖皮质激素,目前研究多集中于它抑制炎性因子分泌而产生镇痛作用,但机制尚不十分清楚。地塞米松能否通过介导P2Y_1受体抑制脊髓背角星形胶质细胞TNF-α的分泌?需要进一步研究。本实验通过纯化培养大鼠脊髓背角星形胶质细胞,观察不同浓度地塞米松对其P2Y_1受体表达的影响,及地塞米松对ADPβs激活后的背角星形胶质细胞P2Y_1受体表达的影响,并通过其对TNF-α分泌的改变推测可能的机制。
方法:培养并纯化大鼠脊髓背角星形胶质细胞,免疫组织化学观察背角星形胶质细胞P2Y_1受体表达水平的变化;ELISA技术检测背角星形胶质细胞TNF-α的分泌。
结果:
第一部分:
1.大鼠脊髓背角星形胶质细胞的培养与纯化,免疫组织化学鉴定细胞类型。结果表明,脊髓背角星形胶质细胞纯度在98%以上。
2.地塞米松呈剂量依赖性抑制脊髓背角星形胶质细胞P2Y_1受体的表达;糖皮质激素受体阻断剂(RU486)特异性阻断糖皮质激素受体后,可拮抗地塞米松对背角星形胶质细胞P2Y_1受体表达的抑制作用。
3.地塞米松呈剂量依赖性抑制脊髓背角星形胶质细胞TNF-α的分泌。RU486特异性阻断糖皮质激素受体后,可拮抗地塞米松对脊髓背角星形胶质细胞TNF-α分泌的抑制作用。
第二部分:
1.ADPβs激活脊髓背角星形胶质细胞,呈剂量依赖性上调P2Y_1受体的表达,在终浓度为100μmol/L时P2Y_1受体表达最多,并促进背角星形胶质细胞TNF-α分泌;P2Y_1受体阻断剂(MRS2179)特异性阻断P2Y_1受体后,可拮抗ADPβs对P2Y_1受体表达及TNF-α分泌的促进作用。
2.ADPβs激活脊髓背角星形胶质细胞,不同浓度地塞米松可呈剂量依赖性的拮抗ADPβs对背角星形胶质细胞P2Y_1受体表达及TNF-α分泌的促进作用。
结论:
1.地塞米松对脊髓背角星形胶质细胞P2Y_1受体表达及TNF-α的分泌均有抑制作用。
2. ADPβs激活背角星形胶质细胞可以使P2Y_1受体表达上调。
3.地塞米松可抑制激活的脊髓背角星形胶质细胞P2Y_1受体的表达及TNF-α的分泌。
综上所述,地塞米松可能通过抑制P2Y_1受体表达和TNF-α的分泌起到提高痛阈的作用;亦可能通过介导P2Y_1受体抑制炎性因子TNF-α的分泌,说明地塞米松在疼痛状态下起到了镇痛作用。
It is known that pain does harm to health. The spinal dorsal horn plays an important role in nociceptive information transmission from the periphery to central nervous system. While neuronal functioning is altered, there is significant evidence showing that exaggerated pain is regulated by the activation of astrocytes and microglia. More and more data suggest that the activation of spinal cord glia is necessary for the development and maintenance of pathological pain and hyperalgesia.Moreover, envidences have revealed that the development and maintenance of pathological pain has something to do with P2 receptor, which take part in nociceptive information transmission.P2Y_1 receptor express at astrocytes which located in spinal dorsal horn, and activation of P2Y_1 recetor leads to the secretion of TNF-α.
Resesrches on dexamethasone have focused on inhibiting inflammatory factor, however mechanism is unknown.Can dexamethasone ihibit the secretion of TNF-αthrough P2Y_1 receptor pathway,and through activating P2Y_1 receptor pathway?This study will use some methods includeing morphology and ELISA to investigate the relationship between dexamethasone and P2Y_1 receptor in cultured rat dorsal horn astrocytes.
Methods: Primary dissociated culture of dorsal spinal cord astrocytes were prepared from Sprague–Dawley rats. The changes of P2Y_1 receptor which expressed in cultured dorsal horn astrocytes were observed by using immunohistochemical staining.And to observe the secretion of TNF-αby means of ELISA.
Results:
Part one:
1. To observe the puration of astrocytes by using immunohistochemical staining,and that of tham was more than 98%.
2. Dexamethasone (0.01~100μmol/L) can dose-dependently decrease the expression of P2Y_1 receptor in cultured rat dorsal horn astrocytes;While after RU486 (1000μmol/L) block the glucocorticoid reccptor, the wok resulted from Dexamethasone become weaker.
3. Dexamethasone (0.01~100μmol/L) can dose-dependently decrease the secretion of TNF-αin cultured rat dorsal horn astrocytes;While after RU486 (100μmol/L) block the glucocorticoid reccptor, the wok resulted from Dexamethasone become weaker.
Part two:
1. ADPβwhich activates cultured dorsal horn atrocytes can dose-dependently increase the expression of P2Y_1 receptor and the secretion of TNF-α; MRS2179(0.1~1000μmol/L), P2Y_1-specific antagonist, can inhibit increasing of the expression of P2Y_1 receptor and the secretion of TNF-α.
2. After ADPβs activated cultured dorsal horn atrocytes, Dexamethasone can dose-dependently the work laeded by ADPβs.
Conclusions:
1. Dexamethasone decrease the expression of P2Y_1 receptor and the secretion of TNF-α.
2. Astrocytes activated by ADPβs can increase the expression of P2Y_1 receptor.
3. Dexamethasone can inhibit the expression of P2Y_1 receptor and the secretion of TNF-αin activated astrocyte.
In summary, these results suggest that Dexamethasone may increase the pain threshold by inhibiting the expression of P2Y_1 receptor and secretion of TNF-αin cultured dorsal horn astrocytes ;We also provide evidence that Dexamethasone can release the pain threshold by inhibiting the secretion of TNF-α.
引文
[1] Sung B,Lim G,Mao J. Altered spinal arachidonic acid turnover after peripheral nerve injury regulates regional glutamate concentration and neuropathic pain behaviors in rats[J]. Pain,2007,131(1-2):121-131.
[2] Campbell JN,Meyer RA. Mechanisms of Neuropathic Pain[J]. Neuron, 2006, 52(1):77-92.
[3]张水文,孙凤岭,臧桐,等.星形胶质细胞活化对慢性前列腺炎疼痛大鼠脑诱发电位的影响[J].中国现代医学杂志, 2008, 18(10):1328-1330 .
[4] JT Porter and KD McCarthy,Astrocytic neurotransmitter receptors in situ and in vivo. Prog Neurobiol, Mar 1997; 51(4): 439-55.
[5] James T. potter and Ken D. Mccarthy, Astrocytic neurotransmitterReceptors[J]. Neurobiology, 1997, 51,439-455. .
[6]马媛媛, SP及其NK-1受体促进大鼠伤害性传入引起的脊髓后角星形胶质细胞的激活,河北医科大学.
[7] DOLY Stéphane ; FISCHER Jacqueline ; SALIO Chiara ; CONRATH Marie ; The vanilloid receptor-1 is expressed in rat spinal dorsal horn astrocytes , Neuroscience letters.
[8] L Hosli, E Hosli, and H Kaser, Colocalization of cholinergic, adrenergic and peptidergic receptors on astrocytes. Neuroreport, June 1, 1993; 4(6): 679-82.
[9]李茂全,王艳艳,杨书,陈朝琼,杨晓虹,余争平,糖皮质激素致星形胶质细胞内钙浓度的快速升高,第四军医大学学报, 2006,26(23),2124-2127.
[10] Jun-wei Zeng,Xiaohong Liu,Jinhai-Zhang,Xigui-Wu,Huai-zhen Ruan,P2Y1 receptor-mediated glutamate relesae from cultured dorsal spinal cord astrocytes,Journal of neurochemistry,2008. 106:2106-2118.
[11] Wang S,Lin G,Yang L,et al. Downregulation of spinal glutamate transporter EAAC1 following nerve injury is regulated by central glucocorticoid receptors in rats[J]. pain 2006,120(1-2):78-85
[12]金保方,黄宇烽,杨晓玉,等.加味枸橘汤治疗慢性盆腔疼痛综合征的临床研究[J] .医学研究生学报, 2005, 18(8):700-701.
[13]孙怡,田玉科,项红兵,氢化可的松对大鼠脊髓星形胶质细胞体外分泌TNF-alpha及分泌EAAs的影响[J].中国现代医学杂志, 2004, 14 (7):103-105.
[14] Brewer JA,Khor B,Vogt SK,Muglia LM,Fujiwara H,Haegele KE,Sleckman BP,Muglia LJ. T-cell glucocorticoid receptor is required to suppress cox-2-mediated lethal immune activation. Nat Med,2003 Oct;9(10):1318-22.
[15] Maria Domercq, Liliana Brambilla, Ethel Pilati, et al . P2Y1 Receptor-evoked Glutamate Exocytosis from Astrocytes control by tumor necrosis factor-αand prostaglandins[J], The Journal Of Biological Chemistry . 2006, 281( 41): 30684–30696.
[16] Ruan HZ, Burnstock G. Localisation of P2Y1 and P2Y4 receptors in dorsal root, nodose and trigeminal ganglia of the rat. Histochem Cell Biol. 2003; 120: 415-426.
[17] Ruan HZ, Birder LA, De Groat WC, et al. Localization of P2X and P2Y receptors in dorsal root ganglia of the cat. J Histochem Cytochem. 2005, 53: 1273-82.
[18] Gallagher CJ, Salter MW. Differential properties of astrocyte calcium waves mediated by P2Y1 and P2Y2 receptors. J Neurosci. 2003; 23: 6728-6739.
[19] Kobayashi K, Fukuoka T, Yamanaka H, Dai Y, Obata K, Tokunaga A, Noguchi K. Neurons and Glial Cells differentially express P2Y receptor mRNAs in the rat dorsal root ganglion and spinal cford. The Journal of comparative Neurology. 2006; 498: 443–454.
[20] Watkins LR, Milligan ED, Maier SF,et al. Glial activation: A driving force for pathological pain[J]. Trends neurosci 2001; 24: 450–5.
[21] Tsuda M, Shigemoto-Mogami Y, Koizumi S,et al. . P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury[J]. Nature 2003; 424: 778–83.
[22] Salter M. W., Hicks J. L. ATP-evoked increases in intracellular calcium in neurons and glia from the dorsal spinal cord [J]. Neurosci. 1994,14, 1563–1575.
[23] Jun-Wei Zeng,1 Xiao-Hong Liu,1,2 Yan-Dong Zhao.Role of P2Y1 Receptor in astroglia-to-Neuron Signaling at Dorsal Spinal Cord. Journal of Neuroscience Research 87:2667–2676 (2009)
[24] Jun-Wei Zeng ,Xiao-Hong Liu , Huai-Zhen Ruan. Inhibition of ATP-Induced Glutamate Release by MRS2179 in Cultured Dorsal Spinal Cord Astrocytes.Pharmacology, 2008;82:257–263.
[25] Kantor P,Lucien A,Kozak R,et al. The antianginal drug trimetazidine shifts cardiac energy merabolism from fatty acid oxidation to glucose oxidation by inhibitingmitochondrial long chain 3-Ketoacyl coenzyme A thiolase. Circ Res,2000;86:580-588
[26]刘晓红,曾俊伟,阮怀珍,等.地塞米松对大鼠背根神经节细胞ATP激活电流的快速抑制作用[J],解放军医学杂志, 2008:5(33), 590-592
[27] Park Y S,Ha-Choi Y,Park C H,et al. Nogenomic glucocorticoid effects on activity-dependent potentiation of catecholamine release in chromaffin cells[J]. Endocrinology,2008,149(10):4921-4927
[28] Han J Z,Liu W,Chen Y Z,et al. Inhibition of ATP -induced calcium influx in HT4 cells by glucocorticoids:involvement of protein kinase A[J]. Acta Pharmacol Sin,2005,26(2): 199-204.
[29]刘晓红,曾俊伟,阮怀珍,等,地塞米松快速抑制大鼠背根神经节细胞中ATP所致的钙升高[J],第三军医学报, 2009, 31(14):1329-1332。
[30] Kim H M, Lee E H, Nah J, et a1.Efect of Poly a tenuifolia root extract on the tumor necrosis factor-alpha secretion from mouse astrocytes[J].J Ethnopharmacol, 1998, 61(3):201-208.
[31] STALDER A K, CARSON M J.PAGENSTECHER A, et a1.Late-on-set chronic inflammatory encephaopathy in immune-competent and severe combined immune- deficient(SCID)mice with astrocyte-targeted expression oftumor necrosisfactor[J]. Am J Pathol, 1998, 153(3):767-783.
[32] Brewer JA,Khor B,Vogt SK, et al. T-cell glucocorticoid receptor is required to euppress cox-2-mediated lethal immune activation. Nature Medicine 9, 1318-1322.
[33] Kim HT,Park SK,et al. Non-noxious A fiber afferent input enhances capsaicin-induced mechanical hyperalgesia in the rat[J]. Pain,2001,94:169-175.
[1] Collier HO, James GW, Schneider C. Antagonism by aspirin and fenamates of bronchoconstriction and nociception induced by adenosine-5S-triphosphate. Nature 1966; 212: 411–2.
[2] Bleehen T, Keele CA. Observations on the algogenic actions of adenosine compounds on the human blister base preparation. Pain 1977; 4: 367–77.
[3] Chen CC, Akopian AN, Sivilotti L et al. A P2X purinoceptor expressed by a subset of sensory neurons. Nature 1995; 377: 428–31.
[4] Lewis C, Neidhart S, Holy C et al. Coexpression of P2X2 and P2X3 receptor subunits can account for ATP-gated currents in sensory neurons. Nature 1995; 377: 432–5.5. Hoffmann, C., Moro, S., Nicholas, R. A., Harden, K. T., and Jacobson,K. A. (1999). J. Biol. Chem. 274: 14639–14647.
[5] Rivera-Arconada I, Martinez-Gomez J, Lopez-Garcia JA, et al. M-current modulators alter rat spinal nociceptive transmission: an electrophysiological study in vitro. Neuropharmacology 2004; 46: 598–606.
[6] Burnstock, G. Purinergic receptors and pain [J]. Curr Pharm Des 2009, 15(15): 1717-35
[7] Takeshi Hiramoto,Wataru Nemoto,Takeshi Kikuchi,and Norihisa Fujita,Construction of Hypothetical Three-Dimensional Structure of P2Y1 Receptor Based on Fourier Transform Analysis Journal of Protein Chemistry, Vol. 21, No. 8, November 2002
[8] Burasteck G.A basis for distinguishing two types of purinergic receptor.In:Cell membrane receptors for drugs and hormones:A muhidisciplinary approach (Straub LB ed).New Y0rk:Raven Press,1978:107-118
[9] von Ku¨gelgen I, Wetter A. Molecular pharmacology of P2Y-receptors. Naunyn-Schmiedeberg’s Arch Pharmacol 2000; 362: 310–23.
[10] Jacobson KA, Costanzi S, Ohno M et al. Molecular recognition at purine and pyrimidine nucleotide (P2) receptors. Curr Top Med Chem 2004; 4: 805–19.
[11] Burnstock G. Introduction: P2 receptors. Curr Top Med Chem 2004;4: 793–803.
[12] Fischer B.P2-receptor agonists:From molecular recognition studies to po tentialclinical applications.Drug Dev Res 2000;50:338-354
[13] lllesa P,Alexandre Ribeiro J.Molecular physiology of P2 receptors in the central nervous system.Eur J Pharmacol,2004;483:5—17
[14] Tominaga M, Wada M, Masu M. Potentiation of capsaicin receptor activity by metabotropic ATP receptors as a possible mechanism for ATP-evoked pain and hyperalgesia. Proc Natl Acad Sci USA 2001;98: 6951–6.
[15] Sanada M, Yasuda H, Omatsu-Kanbe M et al. Increase in intracellular Ca2+ and calcitonin gene-related peptide release through metabotropic P2Y receptors in rat dorsal root ganglion neurons.Neuroscience 2002; 111: 413–22.
[16] Ruan HZ, Burnstock G. Localisation of P2Y1 and P2Y4 receptors in dorsal root, nodose and trigeminal ganglia of the rat. Histochem Cell Biol 2003; 120: 415–26.
[17] Xiao HS, Huang QH, Zhang FX et al. Identification of gene expression profile of dorsal root ganglion in the rat peripheral axotomy model of neuropathic pain. Proc Natl Acad Sci USA 2002;99: 8360–5.
[18] Moriyama T, Iida T, Kobayashi K et al. Possible involvement of P2Y2 metabotropic receptors in ATP-induced transient receptor potential vanilloid receptor 1-mediated thermal hypersensitivity. J Neurosci 2003; 23: 6058–62.
[19] Nakamura F, Strittmatter SM. P2Y1 purinergic receptors in sensory neurons: Contribution to touch-induced impulse generation. Proc Natl Acad Sci USA 1996; 93: 10465–70.
[20] Fong AY, Krstew EV, Barden J, Lawrence AJ. Immunoreactive localisation of P2Y1 receptors within the rat and human nodose ganglia and rat brainstem: Comparison with [a33P]deoxyadenosine 5S-triphosphate autoradiography. Neuroscience 2002; 113: 809–23.
[21] Gerevich Z, Borvendeg SJ, Schro¨der W et al. Inhibition of N-type voltage-activated calcium channels in rat dorsal root ganglion neurons by P2Y receptors is a possible mechanism of ADP-induced analgesia. J Neurosci 2004; 24: 797–807
[22] Borvendeg SJ, Gerevich Z, Gillen C, Illes P. P2Y receptor-mediated inhibition of voltage-dependent Ca2+ channels in rat dorsal root ganglion neurons. Synapse 2003;47: 159–61.
[23] Pannese E. The satellite cells of the sensory ganglia. Adv Anat Embryol Cell Biol 1981; 65: 1–111.
[24] Woodham P, Anderson PN, Nadim W, Turmaine M. Satellite cells surrounding axotomised rat dorsal root ganglion cells increase expression of a GFAP-like protein. Neurosci Lett 1989; 98: 8–12.
[25] Stephenson JL, Byers MR. GFAP immunoreactivity in trigeminal ganglion satellite cells after tooth injury in rats. Exp Neurol 1995;131: 11–22.
[26] Li L, Zhou XF. Pericellular Griffonia simplicifolia I isolectin B4-binding ring structures in the dorsal root ganglia following peripheral nerve injury in rats. J Comp Neurol 2001; 439: 259–74.
[27] Hanani M, Huang TY, Cherkas PS et al. Glial cell plasticity in sensory ganglia induced by nerve damage. Neuroscience 2002; 114:279–83.
[28] Weick M, Cherkas PS, Hartig W et al. P2 receptors in satellite glial cells in trigeminal ganglia of mice. Neuroscience 2003; 120: 969–77.
[29] Salter MW, Hicks JL. ATP-evoked increases in intracellular calcium in neurons and glia from the dorsal spinal cord. J Neurosci 1994; 14:1563–75.
[30] Fam SR, Gallagher CJ, Salter MW. P2Y1 purinoceptor-mediated Ca2+ signaling and Ca2+ wave propagation in dorsal spinal cord astrocytes. J Neurosci 2000; 20: 2800–8.
[31] Ho C, Hicks J, Salter MW. A novel P2-purinoceptor expressed by a subpopulation of astrocytes from the dorsal spinal cord of the rat. Br J Pharmacol 1995; 116: 2909–18.
[32] Idestrup CP, Salter MW. P2Y and P2U receptors differentially release intracellular Ca2+ via the phospholipase cinositol 1,4,5-triphosphate pathway in astrocytes from the dorsal spinal cord.Neuroscience 1998; 86: 913–23.
[33] Jun-wei Zeng,Xiaohong Liu,Jinhai-Zhang,Xigui-Wu,Huai-zhen Ruan,P2Y1 receptor-mediated glutamate relesae from cultured dorsal spinal cord astrocytes,Journal of neurochemistry,2008.106:2106-2118
[34]曾俊伟,刘晓红,阮怀珍.ATP通过P2Y1受体引发脊髓背角星形胶质细胞[Ca2+]i升高和GFAP表达上调.免疫学杂志第2,4卷第2期2OO8年3月
[35] Gallagher CJ, Salter MW. Differential properties of astrocyte calcium waves mediated by P2Y1 and P2Y2 receptors. J Neurosci 2003; 23: 6728–39.
[36] Pharmacol Rev,Burnstock G.Receptor for purines and pyrimidines.Pharmacol Rev,1998,50(3):413-492
[37] Bowden A,Patel V,Brown C,et al.Evidence for requirement of tyrosine phosphory-lation in endothelial P2Y-and P2u-purinoceptoe stimulation of prostacyclin release.Br J Pharmacol,1995,116(6):2563-2568.
[38] PatelV,Brown C,Goodwin A,et al.Phoosphorylation and activation of P32 and P44 mitogen-activated protein kinase are required for the P2 purinoceptor stimulation of endothelial prostacyclin production.Biochem J,1996,320(1):221-226.
[39] Berti-Mattera LN,wilkins PL,Mahum Z,et al.P2-purinergic receptors regulate phosphlipase C and adenylate cyclase activatise in immortalized Schwann cell.Biochem J.1996,314(2):555-561.
[40] Webb TE,Feolde E,Vigne P,et al.The P2Yreceptors in rat brain microvascular endothelial cells couple to inhibition of adenylate cyclase.Br J Pharmacol, 1996,119:1385-1392.
[41] Ikeuch Y,Nishizaki.P2-purinoceptor-orperated postassium channel in rat cerebellar neurons.Biochem Biophys Res Commum,1996,218(1):67-71
[42] Usachev YM, DeMarco SJ, Campbell C et al. Bradykinin and ATP accelerate Ca2+ efflux from rat sensory neurons via protein kinase C and the plasma membrane Ca2+ pump isoform 4. Neuron 2002; 33:113–22.
[43] Watkins LR, Milligan ED, Maier SF. Glial activation: A driving force for pathological pain. Trends Neurosci 2001; 24: 450–5.
[44] Tsuda M, Shigemoto-Mogami Y, Koizumi S et al. P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury.Nature 2003; 424: 778–83.
[45] Safiulina V,Mzalov R,Khiroug L et a1.Reactive oxygen species med iate the po tentiating efects of ATP on GABAergic synaptic transmission in the immature hippocam pus.J Biol Chem,2006;281:23464—23470
[46] Kucher BM,Neary JT.Bi.functional effects of ATP/P2 receptor activation on tumornecrosis factor·alpha release in lipo po lysaccharide stimulated astrocytes . J Neurechem,2005;92:525—535
[47] Maria Domercq, Liliana Brambilla, Ethel Pilati, et al .P2Y1 Receptor-evoked Glutamate Exocytosis from Astrocytes control by tumor necrosis factor-αand prostaglandins[J]. The Journal Of Biological Chemistry .2006 , 281( 41) : 30684–30696.
[48]施维维,何文娟,欧珊,等.地塞米松抑制星形胶质细胞P2Y1R表达及TNF-α分泌.第三军医大学学报,2010,32卷第1期,9-12.
[2] Campbell JN,Meyer RA. Mechanisms of Neuropathic Pain[J]. Neuron, 2006, 52(1):77-92.
[3]张水文,孙凤岭,臧桐,等.星形胶质细胞活化对慢性前列腺炎疼痛大鼠脑诱发电位的影响[J].中国现代医学杂志, 2008, 18(10):1328-1330 .
[4] JT Porter and KD McCarthy,Astrocytic neurotransmitter receptors in situ and in vivo. Prog Neurobiol, Mar 1997; 51(4): 439-55.
[5] James T. potter and Ken D. Mccarthy, Astrocytic neurotransmitterReceptors[J]. Neurobiology, 1997, 51,439-455. .
[6]马媛媛, SP及其NK-1受体促进大鼠伤害性传入引起的脊髓后角星形胶质细胞的激活,河北医科大学.
[7] DOLY Stéphane ; FISCHER Jacqueline ; SALIO Chiara ; CONRATH Marie ; The vanilloid receptor-1 is expressed in rat spinal dorsal horn astrocytes , Neuroscience letters.
[8] L Hosli, E Hosli, and H Kaser, Colocalization of cholinergic, adrenergic and peptidergic receptors on astrocytes. Neuroreport, June 1, 1993; 4(6): 679-82.
[9]李茂全,王艳艳,杨书,陈朝琼,杨晓虹,余争平,糖皮质激素致星形胶质细胞内钙浓度的快速升高,第四军医大学学报, 2006,26(23),2124-2127.
[10] Jun-wei Zeng,Xiaohong Liu,Jinhai-Zhang,Xigui-Wu,Huai-zhen Ruan,P2Y1 receptor-mediated glutamate relesae from cultured dorsal spinal cord astrocytes,Journal of neurochemistry,2008. 106:2106-2118.
[11] Wang S,Lin G,Yang L,et al. Downregulation of spinal glutamate transporter EAAC1 following nerve injury is regulated by central glucocorticoid receptors in rats[J]. pain 2006,120(1-2):78-85
[12]金保方,黄宇烽,杨晓玉,等.加味枸橘汤治疗慢性盆腔疼痛综合征的临床研究[J] .医学研究生学报, 2005, 18(8):700-701.
[13]孙怡,田玉科,项红兵,氢化可的松对大鼠脊髓星形胶质细胞体外分泌TNF-alpha及分泌EAAs的影响[J].中国现代医学杂志, 2004, 14 (7):103-105.
[14] Brewer JA,Khor B,Vogt SK,Muglia LM,Fujiwara H,Haegele KE,Sleckman BP,Muglia LJ. T-cell glucocorticoid receptor is required to suppress cox-2-mediated lethal immune activation. Nat Med,2003 Oct;9(10):1318-22.
[15] Maria Domercq, Liliana Brambilla, Ethel Pilati, et al . P2Y1 Receptor-evoked Glutamate Exocytosis from Astrocytes control by tumor necrosis factor-αand prostaglandins[J], The Journal Of Biological Chemistry . 2006, 281( 41): 30684–30696.
[16] Ruan HZ, Burnstock G. Localisation of P2Y1 and P2Y4 receptors in dorsal root, nodose and trigeminal ganglia of the rat. Histochem Cell Biol. 2003; 120: 415-426.
[17] Ruan HZ, Birder LA, De Groat WC, et al. Localization of P2X and P2Y receptors in dorsal root ganglia of the cat. J Histochem Cytochem. 2005, 53: 1273-82.
[18] Gallagher CJ, Salter MW. Differential properties of astrocyte calcium waves mediated by P2Y1 and P2Y2 receptors. J Neurosci. 2003; 23: 6728-6739.
[19] Kobayashi K, Fukuoka T, Yamanaka H, Dai Y, Obata K, Tokunaga A, Noguchi K. Neurons and Glial Cells differentially express P2Y receptor mRNAs in the rat dorsal root ganglion and spinal cford. The Journal of comparative Neurology. 2006; 498: 443–454.
[20] Watkins LR, Milligan ED, Maier SF,et al. Glial activation: A driving force for pathological pain[J]. Trends neurosci 2001; 24: 450–5.
[21] Tsuda M, Shigemoto-Mogami Y, Koizumi S,et al. . P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury[J]. Nature 2003; 424: 778–83.
[22] Salter M. W., Hicks J. L. ATP-evoked increases in intracellular calcium in neurons and glia from the dorsal spinal cord [J]. Neurosci. 1994,14, 1563–1575.
[23] Jun-Wei Zeng,1 Xiao-Hong Liu,1,2 Yan-Dong Zhao.Role of P2Y1 Receptor in astroglia-to-Neuron Signaling at Dorsal Spinal Cord. Journal of Neuroscience Research 87:2667–2676 (2009)
[24] Jun-Wei Zeng ,Xiao-Hong Liu , Huai-Zhen Ruan. Inhibition of ATP-Induced Glutamate Release by MRS2179 in Cultured Dorsal Spinal Cord Astrocytes.Pharmacology, 2008;82:257–263.
[25] Kantor P,Lucien A,Kozak R,et al. The antianginal drug trimetazidine shifts cardiac energy merabolism from fatty acid oxidation to glucose oxidation by inhibitingmitochondrial long chain 3-Ketoacyl coenzyme A thiolase. Circ Res,2000;86:580-588
[26]刘晓红,曾俊伟,阮怀珍,等.地塞米松对大鼠背根神经节细胞ATP激活电流的快速抑制作用[J],解放军医学杂志, 2008:5(33), 590-592
[27] Park Y S,Ha-Choi Y,Park C H,et al. Nogenomic glucocorticoid effects on activity-dependent potentiation of catecholamine release in chromaffin cells[J]. Endocrinology,2008,149(10):4921-4927
[28] Han J Z,Liu W,Chen Y Z,et al. Inhibition of ATP -induced calcium influx in HT4 cells by glucocorticoids:involvement of protein kinase A[J]. Acta Pharmacol Sin,2005,26(2): 199-204.
[29]刘晓红,曾俊伟,阮怀珍,等,地塞米松快速抑制大鼠背根神经节细胞中ATP所致的钙升高[J],第三军医学报, 2009, 31(14):1329-1332。
[30] Kim H M, Lee E H, Nah J, et a1.Efect of Poly a tenuifolia root extract on the tumor necrosis factor-alpha secretion from mouse astrocytes[J].J Ethnopharmacol, 1998, 61(3):201-208.
[31] STALDER A K, CARSON M J.PAGENSTECHER A, et a1.Late-on-set chronic inflammatory encephaopathy in immune-competent and severe combined immune- deficient(SCID)mice with astrocyte-targeted expression oftumor necrosisfactor[J]. Am J Pathol, 1998, 153(3):767-783.
[32] Brewer JA,Khor B,Vogt SK, et al. T-cell glucocorticoid receptor is required to euppress cox-2-mediated lethal immune activation. Nature Medicine 9, 1318-1322.
[33] Kim HT,Park SK,et al. Non-noxious A fiber afferent input enhances capsaicin-induced mechanical hyperalgesia in the rat[J]. Pain,2001,94:169-175.
[1] Collier HO, James GW, Schneider C. Antagonism by aspirin and fenamates of bronchoconstriction and nociception induced by adenosine-5S-triphosphate. Nature 1966; 212: 411–2.
[2] Bleehen T, Keele CA. Observations on the algogenic actions of adenosine compounds on the human blister base preparation. Pain 1977; 4: 367–77.
[3] Chen CC, Akopian AN, Sivilotti L et al. A P2X purinoceptor expressed by a subset of sensory neurons. Nature 1995; 377: 428–31.
[4] Lewis C, Neidhart S, Holy C et al. Coexpression of P2X2 and P2X3 receptor subunits can account for ATP-gated currents in sensory neurons. Nature 1995; 377: 432–5.5. Hoffmann, C., Moro, S., Nicholas, R. A., Harden, K. T., and Jacobson,K. A. (1999). J. Biol. Chem. 274: 14639–14647.
[5] Rivera-Arconada I, Martinez-Gomez J, Lopez-Garcia JA, et al. M-current modulators alter rat spinal nociceptive transmission: an electrophysiological study in vitro. Neuropharmacology 2004; 46: 598–606.
[6] Burnstock, G. Purinergic receptors and pain [J]. Curr Pharm Des 2009, 15(15): 1717-35
[7] Takeshi Hiramoto,Wataru Nemoto,Takeshi Kikuchi,and Norihisa Fujita,Construction of Hypothetical Three-Dimensional Structure of P2Y1 Receptor Based on Fourier Transform Analysis Journal of Protein Chemistry, Vol. 21, No. 8, November 2002
[8] Burasteck G.A basis for distinguishing two types of purinergic receptor.In:Cell membrane receptors for drugs and hormones:A muhidisciplinary approach (Straub LB ed).New Y0rk:Raven Press,1978:107-118
[9] von Ku¨gelgen I, Wetter A. Molecular pharmacology of P2Y-receptors. Naunyn-Schmiedeberg’s Arch Pharmacol 2000; 362: 310–23.
[10] Jacobson KA, Costanzi S, Ohno M et al. Molecular recognition at purine and pyrimidine nucleotide (P2) receptors. Curr Top Med Chem 2004; 4: 805–19.
[11] Burnstock G. Introduction: P2 receptors. Curr Top Med Chem 2004;4: 793–803.
[12] Fischer B.P2-receptor agonists:From molecular recognition studies to po tentialclinical applications.Drug Dev Res 2000;50:338-354
[13] lllesa P,Alexandre Ribeiro J.Molecular physiology of P2 receptors in the central nervous system.Eur J Pharmacol,2004;483:5—17
[14] Tominaga M, Wada M, Masu M. Potentiation of capsaicin receptor activity by metabotropic ATP receptors as a possible mechanism for ATP-evoked pain and hyperalgesia. Proc Natl Acad Sci USA 2001;98: 6951–6.
[15] Sanada M, Yasuda H, Omatsu-Kanbe M et al. Increase in intracellular Ca2+ and calcitonin gene-related peptide release through metabotropic P2Y receptors in rat dorsal root ganglion neurons.Neuroscience 2002; 111: 413–22.
[16] Ruan HZ, Burnstock G. Localisation of P2Y1 and P2Y4 receptors in dorsal root, nodose and trigeminal ganglia of the rat. Histochem Cell Biol 2003; 120: 415–26.
[17] Xiao HS, Huang QH, Zhang FX et al. Identification of gene expression profile of dorsal root ganglion in the rat peripheral axotomy model of neuropathic pain. Proc Natl Acad Sci USA 2002;99: 8360–5.
[18] Moriyama T, Iida T, Kobayashi K et al. Possible involvement of P2Y2 metabotropic receptors in ATP-induced transient receptor potential vanilloid receptor 1-mediated thermal hypersensitivity. J Neurosci 2003; 23: 6058–62.
[19] Nakamura F, Strittmatter SM. P2Y1 purinergic receptors in sensory neurons: Contribution to touch-induced impulse generation. Proc Natl Acad Sci USA 1996; 93: 10465–70.
[20] Fong AY, Krstew EV, Barden J, Lawrence AJ. Immunoreactive localisation of P2Y1 receptors within the rat and human nodose ganglia and rat brainstem: Comparison with [a33P]deoxyadenosine 5S-triphosphate autoradiography. Neuroscience 2002; 113: 809–23.
[21] Gerevich Z, Borvendeg SJ, Schro¨der W et al. Inhibition of N-type voltage-activated calcium channels in rat dorsal root ganglion neurons by P2Y receptors is a possible mechanism of ADP-induced analgesia. J Neurosci 2004; 24: 797–807
[22] Borvendeg SJ, Gerevich Z, Gillen C, Illes P. P2Y receptor-mediated inhibition of voltage-dependent Ca2+ channels in rat dorsal root ganglion neurons. Synapse 2003;47: 159–61.
[23] Pannese E. The satellite cells of the sensory ganglia. Adv Anat Embryol Cell Biol 1981; 65: 1–111.
[24] Woodham P, Anderson PN, Nadim W, Turmaine M. Satellite cells surrounding axotomised rat dorsal root ganglion cells increase expression of a GFAP-like protein. Neurosci Lett 1989; 98: 8–12.
[25] Stephenson JL, Byers MR. GFAP immunoreactivity in trigeminal ganglion satellite cells after tooth injury in rats. Exp Neurol 1995;131: 11–22.
[26] Li L, Zhou XF. Pericellular Griffonia simplicifolia I isolectin B4-binding ring structures in the dorsal root ganglia following peripheral nerve injury in rats. J Comp Neurol 2001; 439: 259–74.
[27] Hanani M, Huang TY, Cherkas PS et al. Glial cell plasticity in sensory ganglia induced by nerve damage. Neuroscience 2002; 114:279–83.
[28] Weick M, Cherkas PS, Hartig W et al. P2 receptors in satellite glial cells in trigeminal ganglia of mice. Neuroscience 2003; 120: 969–77.
[29] Salter MW, Hicks JL. ATP-evoked increases in intracellular calcium in neurons and glia from the dorsal spinal cord. J Neurosci 1994; 14:1563–75.
[30] Fam SR, Gallagher CJ, Salter MW. P2Y1 purinoceptor-mediated Ca2+ signaling and Ca2+ wave propagation in dorsal spinal cord astrocytes. J Neurosci 2000; 20: 2800–8.
[31] Ho C, Hicks J, Salter MW. A novel P2-purinoceptor expressed by a subpopulation of astrocytes from the dorsal spinal cord of the rat. Br J Pharmacol 1995; 116: 2909–18.
[32] Idestrup CP, Salter MW. P2Y and P2U receptors differentially release intracellular Ca2+ via the phospholipase cinositol 1,4,5-triphosphate pathway in astrocytes from the dorsal spinal cord.Neuroscience 1998; 86: 913–23.
[33] Jun-wei Zeng,Xiaohong Liu,Jinhai-Zhang,Xigui-Wu,Huai-zhen Ruan,P2Y1 receptor-mediated glutamate relesae from cultured dorsal spinal cord astrocytes,Journal of neurochemistry,2008.106:2106-2118
[34]曾俊伟,刘晓红,阮怀珍.ATP通过P2Y1受体引发脊髓背角星形胶质细胞[Ca2+]i升高和GFAP表达上调.免疫学杂志第2,4卷第2期2OO8年3月
[35] Gallagher CJ, Salter MW. Differential properties of astrocyte calcium waves mediated by P2Y1 and P2Y2 receptors. J Neurosci 2003; 23: 6728–39.
[36] Pharmacol Rev,Burnstock G.Receptor for purines and pyrimidines.Pharmacol Rev,1998,50(3):413-492
[37] Bowden A,Patel V,Brown C,et al.Evidence for requirement of tyrosine phosphory-lation in endothelial P2Y-and P2u-purinoceptoe stimulation of prostacyclin release.Br J Pharmacol,1995,116(6):2563-2568.
[38] PatelV,Brown C,Goodwin A,et al.Phoosphorylation and activation of P32 and P44 mitogen-activated protein kinase are required for the P2 purinoceptor stimulation of endothelial prostacyclin production.Biochem J,1996,320(1):221-226.
[39] Berti-Mattera LN,wilkins PL,Mahum Z,et al.P2-purinergic receptors regulate phosphlipase C and adenylate cyclase activatise in immortalized Schwann cell.Biochem J.1996,314(2):555-561.
[40] Webb TE,Feolde E,Vigne P,et al.The P2Yreceptors in rat brain microvascular endothelial cells couple to inhibition of adenylate cyclase.Br J Pharmacol, 1996,119:1385-1392.
[41] Ikeuch Y,Nishizaki.P2-purinoceptor-orperated postassium channel in rat cerebellar neurons.Biochem Biophys Res Commum,1996,218(1):67-71
[42] Usachev YM, DeMarco SJ, Campbell C et al. Bradykinin and ATP accelerate Ca2+ efflux from rat sensory neurons via protein kinase C and the plasma membrane Ca2+ pump isoform 4. Neuron 2002; 33:113–22.
[43] Watkins LR, Milligan ED, Maier SF. Glial activation: A driving force for pathological pain. Trends Neurosci 2001; 24: 450–5.
[44] Tsuda M, Shigemoto-Mogami Y, Koizumi S et al. P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury.Nature 2003; 424: 778–83.
[45] Safiulina V,Mzalov R,Khiroug L et a1.Reactive oxygen species med iate the po tentiating efects of ATP on GABAergic synaptic transmission in the immature hippocam pus.J Biol Chem,2006;281:23464—23470
[46] Kucher BM,Neary JT.Bi.functional effects of ATP/P2 receptor activation on tumornecrosis factor·alpha release in lipo po lysaccharide stimulated astrocytes . J Neurechem,2005;92:525—535
[47] Maria Domercq, Liliana Brambilla, Ethel Pilati, et al .P2Y1 Receptor-evoked Glutamate Exocytosis from Astrocytes control by tumor necrosis factor-αand prostaglandins[J]. The Journal Of Biological Chemistry .2006 , 281( 41) : 30684–30696.
[48]施维维,何文娟,欧珊,等.地塞米松抑制星形胶质细胞P2Y1R表达及TNF-α分泌.第三军医大学学报,2010,32卷第1期,9-12.