Rapid Elevation of Calcium Concentration in Cultured Dorsal Spinal Cord Astrocytes by Corticosterone
Rapid Elevation of Calcium Concentration in Cultured Dorsal Spinal Cord Astrocytes by Corticosterone
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皮质酮快速升高培养的背脊髓星形胶质细胞中的钙浓度
DOI：
10.1007/s11064-012-0929-4
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发表时间：
2013-02
期刊：
影响因子：
--
通讯作者：
刘晓红
中科院分区：
文献类型：
--
作者：
刘晓红
除了经典的基因组效应外，越来越多的证据表明，GC可以通过非基因组途径对许多组织和细胞产生多种快速效应。用激光共聚焦扫描显微镜观察皮质酮(Cort)对培养的脊髓背侧星形胶质细胞[Ca~(2+)]_i的影响，用荧光指示剂Fluo-4/AM实时监测[Ca~(2+)]_i的变化。皮质酮(CORT，0.01~10μM)可引起培养的脊髓星形胶质细胞[Ca~(2+)]_i迅速升高，并呈剂量依赖关系。百日咳毒素(G蛋白激活阻断剂，100 ng/ml)可阻断皮质醇对星形胶质细胞[Ca~(2+)]i的作用，而糖皮质激素受体拮抗剂RU38486(10μM)对其无影响。此外，放线菌酮(蛋白质合成抑制剂，10μg/ml)预处理不能抑制皮质醇诱导的[Ca~(2+)]i升高。此外，氯化白屈菜红碱(蛋白激酶C抑制剂，10μM)可阻断CORT诱导的钙动员，而蛋白激酶A抑制剂H89(10μM)对此无明显影响。这些观察结果提示，皮质醇对培养的脊髓背侧星形胶质细胞[Ca~(2+)]_i的影响可能与非基因组途径有关。此外，我们的结果还提出了一种可能性，即可能是百日咳毒素敏感的MGCR(G蛋白偶联膜结合型糖皮质激素受体)和蛋白激酶C的下游激活可能与皮质醇诱导培养的脊髓背侧星形胶质细胞钙动员有关。
In addition to the classic genomic effects, increasing evidence suggests that GC can generate multiple rapid effects on many tissues and cells through nongenomic pathway. In the present study, the effects of corticosterone (CORT) on the intracellular calcium concentration ([Ca2+]i) in cultured dorsal spinal cord astrocytes were detected with confocal laser scanning microscopy using fluo-4/AM as a calcium fluorescent indicator that could monitor real-time alterations of [Ca2+]i. CORT (0.01–10 μM) caused a rapid increase in [Ca2+]i with a dose-dependent manner in cultured dorsal spinal cord astrocytes. The action of CORT on astrocytic [Ca2+]i was blocked by pertussis toxin (a blocker of G protein activation, 100 ng/ml), but was unaffected by RU38486 (glucocorticoid receptor antagonist, 10 μM). In addition, cycloheximide (protein-synthesis inhibitor, 10 μg/ml) pretreatment could not impair the CORT-evoked [Ca2+]i elevation. Furthermore, Ca2+mobilization induced by CORT was abolished by chelerythrine chloride (protein kinase C inhibitor, 10 μM), but was not impaired by H89 (protein kinase A inhibitor, 10 μM). These observations suggest that a nongenomic pathways might be involved in the effect of CORT on [Ca2+]i in cultured dorsal spinal cord astrocytes. In addition, our results also raise a possibility that a putative pertussis toxin-sensitive mGCR (G-protein-coupled membrane-bound glucocorticoid receptor) and the downstream activation of protein kinase C may be responsible for CORT-induced Ca2+mobilization in cultured dorsal spinal cord astrocytes.
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