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目的:观察ADP对培养的脊髓背角小胶质细胞内Ca2+浓度([Ca2+]i)的影响及作用机制。方法:培养纯化新生SD大鼠脊髓背角小胶质细胞,免疫组化观察P2Y13受体表达,激光共聚焦显微镜检测ADP作用下,小胶质细胞[Ca2+]i的变化。结果:大鼠脊髓背角小胶质细胞表达P2Y13受体,ADP可导致培养的脊髓背角小胶质细胞[Ca2+]i快速增高,且呈现剂量依赖性;P2Y13受体拮抗剂MRS2211(100μmol/L)能基本阻断ADP的作用,而P2Y1受体拮抗剂MRS2179(100μmol/L)、P2Y12受体拮抗剂MRS2395(100μmol/L)均不影响ADP致小胶质细胞[Ca2+]i升高的效应。结论:ADP可能通过P2Y13受体途径,导致培养的脊髓背角小胶质细胞[Ca2+]快速升高。
Objective: To investigate the effect of ADP on the intracellular Ca2 + concentration ([Ca2 +] i) in cultured spinal dorsal horn microglia and its mechanism. Methods: Purified spinal cord microdialysis cells of neonatal SD rats were cultured and the expression of P2Y13 receptor was observed by immunohistochemistry. The change of [Ca2 +] i in microglia was detected by laser confocal microscopy. Results: P2Y13 receptor was expressed in the dorsal horn microglia of rat spinal cord. ADP induced a rapid increase of [Ca2 +] i in the dorsal horn microglia of spinal cord in a dose-dependent manner. P2Y13 receptor antagonist MRS2211 (100μmol / L) could block the action of ADP, while P2Y1 receptor antagonist MRS2179 (100μmol / L) and P2Y12 receptor antagonist MRS2395 (100μmol / L) did not affect the ADP-induced increase of [Ca2 +] i in microglia effect. CONCLUSION: ADP may pass through the P2Y13 receptor pathway, resulting in a rapid increase of [Ca2 +] in cultured spinal dorsal horn microglia.