摘要
目的观察体外培养的脊髓背角星形胶质细胞P2X1-7受体表达,以及不同浓度三磷酸腺苷(adenosine triphosphate,ATP)对P2X1-7受体及胶质纤维酸性蛋白(glialfibrillary acidic protein,GFAP)表达的影响。方法培养并纯化脊髓背角星形胶质细胞,采用免疫组织化学染色观察P2受体表达。随后在无血清培养条件下,加入不同浓度ATP(100μM,500μM,1mM,2mM)作用24h后,观察脊髓背角星形胶质细胞P2X1-7受体表达及GFAP表达的变化。对照组加入0.01MPBS处理24h。IPP图象分析软件得到P2X受体以及GFAP阳性细胞平均光密度。结果体外培养的大鼠脊髓背角星形胶质细胞表达P2X1,P2X2,P2X4,P2X5,P2X6和P2X7受体;在ATP(100μM,500μM,1mM,2mM)作用下,P2X1,P2X2,P2X4,P2X5,P2X6受体表达未见明显变化,而P2X7受体和GFAP表达逐渐上调,并具量效关系。结论体外培养的大鼠脊髓背角星形胶质细胞表达P2X1,P2X2,P2X4,P2X5,P2X6和P2X7受体,而ATP可以诱发星形胶质细胞GFAP及P2X7受体表达上调,这一结果提示,其中P2X7可能参与脊髓背角星形胶质细胞活性的调节。
Objective To observe the expression of P2X receptor in cultured spinal cord dorsal horn astrocytes,and to assess the effect of adenosine triphosphate（ATP） concentration on the expression of P2X receptor and GFAP. Methods Astrocytes were cultured in free serum medium mixed with different concentrations of ATP （100μM, 500μM, 1 mM, 2mM） for 24 hours. The control group was dealt with 0. 01M PBS for 24 hours. The expression of GFAP and P2X1-7 receptor in cultured astrocytes were observed by using immunohistochemical staining. Image-Pro Plus （IPP） system was used to obtain the number and average optical density （AOD） of the expression of GFAP and P2X1-7 receptor. Results Immunohistochemical results showed the expression of P2X1 , P2X2, P2X4, P2X5, P2X6 and P2X7 receptor protein in cultured dorsal horn astrocytes. The expression of GFAP increased with the increase of ATP concentration. ATP （500μM, 1mM, 2mM） significantly increased the expression of P2X7 receptor. However, the expression of P2X1 , P2X2, P2X4, P2X5, P2X6 receptors was not influenced by ATP. Conclusion The cultured spinal cord dorsal horn astrocytes expressed P2X1 , P2X2, P2X4,P2X5 ,P2X6 and P2X7 receptor. ATP caused the activation of cultured astrocytes. The expression of P2X7 receptor increased after ATP administration, which suggested the participation of P2X7 in ATP-evoked activation.
出处
《华南国防医学杂志》
CAS
2008年第3期1-5,共5页
Military Medical Journal of South China
基金
国家自然科学基金项目(30470557
30672022)
