地塞米松对大鼠脊髓背角星形胶质细胞P2Y1受体 - 第三军医大学学报.DOCVIP

地塞米松抑制星形胶质细胞P2Y1R表达及TNF-α分泌 施维维 阮怀珍* (第三军医大学基础医学部神经生物学教研室，重庆市神经科学研究所，重庆，400038) 摘要：目的 观察不同浓度地塞米松对大鼠体外培养脊髓背角星形胶质细胞P2Y1受体表达及TNF-α分泌的影响。方法 培养并纯化大鼠脊髓背角星形胶质细胞，地塞米松（终浓度分别为0.1μmol/L、1μmol/L、10μmol/L）处理24h后，采用免疫组织化学染色观察P2Y1受体的表达，双抗夹心间接酶联免疫吸附法测定上清液中TNF-α含量。结果 地塞米松（0.1μmol/L、1μmol/L、10μmol/L）作用24h后，与对照组比较，脊髓背角星形胶质细胞P2Y1受体表达下调，并呈剂量依赖性（P0.01或 P0.001），TNF-α分泌量亦呈剂量依赖性下降（P0.01或 P0.001）。结论 地塞米松抑制体外培养大鼠脊髓背角星形胶质细胞P2Y1受体表达及TNF-α的分泌。 关键字：脊髓背角；星形胶质细胞；P2Y1 受体；TNF-α 中图法分类号：R282.71 文献标识码：A THE CHANGES OF P2Y1 RECEPTOR TNF-α IN CULTURAL ASTROCYTES OF SPINAL DORSAL HORN INDUCED BY DEXAMTHASONE SHI Wei-wei, DU Lu,HE Wen-juan,OU Shan,CUI Jian,RUAN Huai-zhen (Department of Neurobiology, The Third Military Medical University, Chongqing Institute of Neuroscience,Chongqing,400038) Abstract: Objective To investigate the influence of Dexamethasone(DEX) on the changes of P2Y1R and secretory of TNF-α in cultural astrocytes of spinal dorsal horn. Methods The expression of P2Y1R in astrocytes and the concentration of TNF-α were detected by using immunohistochemical staining and ELISA. Astrocytes were classified into 4 groups: Control, final concentration of Dexamethasone(DEX) at 0.1μmol/L、1μmol/L、10μmol/L. Result We found that the expression of P2Y1R was down-regulated by increasing of Dexamethasone(DEX) final concentration comparing with Control（P0.01 or P0.001） , and the degrees of the TNF-α concentration is paralleling to it（P0.01 or P0.001）. Conclusion The series of results indicated that the raise of the Dexamethasone(DEX) dose can obviously inhibit the expression of P2Y1 receptors and secretory of TNF-α in cultural astrocytes of spinal dorsal horn. KEY WORDS：spinal dorsal horn;astrocytes;P2Y1R; TNF-α  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 [1]，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[2、3]。以往研究发现疼痛的发生和维持与嘌呤类P2 受体有关[4]， 近年研究提示，嘌呤类P2Y受体参与伤害性信息传递TNF-α的分泌未见 相关报道。因此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P2Y1受体表达的影响，并通过对TNF-α分泌的改变推测其可能的机