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目的研究糖皮质激素醋酸可的松对大鼠下丘脑-垂体-肾上腺轴(HPAA)游离钙(Ca2+)及钙调素依赖的蛋白激酶Ⅱ(CaMPKⅡ)的影响。方法用流式细胞仪及钙荧光探针Flou-3/AM来测定细胞中的Ca2+,通过液体闪烁仪测定γ-32P-ATP参入反应底物的摩尔数,用来表示蛋白激酶的活性。结果醋酸可的松组大鼠下丘脑细胞[Ca2+]i荧光强度为129.32±17.74,显著高于对照组60.65±10.64,P<0.05。而醋酸可的松组大鼠下丘脑、肾上腺组织中CaMPKⅡ活性分别为14.07+3.15和3.87+1.47(nmolPi.min-1.mg-1protein),与对照组的7.91+3.2和2.23+0.52(nmolPi.min-1.mg-1protein)相比显著升高,差异有统计学意义,P<0.001。结论糖皮质激素对下丘脑-垂体-肾上腺轴的调控作用与Ca2+、CaMPKⅡ密切相关。
Objective To investigate the effect of glucocorticoids cortisone on calcium (Ca2 +) and calmodulin-dependent protein kinase Ⅱ (CaMPK Ⅱ) in rat hypothalamus-pituitary-adrenal axis (HPAA). Methods Flow cytometry and calcium fluorescence probe Flou-3 / AM were used to determine Ca2 + in cells. The molar number of γ-32P-ATP involved in the reaction was determined by liquid scintillation to express the activity of protein kinase. Results [Ca2 +] i fluorescence intensity of rat cortisone acetate group was 129.32 ± 17.74, significantly higher than that of control group 60.65 ± 10.64, P <0.05. However, the activity of CaMPKⅡin the hypothalamus and adrenal tissue of cortisone acetate group was 14.07 ± 3.15 and 3.87 ± 1.47 nmolPi · min-1.mg-1protein, respectively, which was significantly higher than that of 7.91 ± 3.2 and 2.23 ± 0.52 (nmolPi .min-1.mg-1protein) was significantly higher, the difference was statistically significant, P <0.001. Conclusion The regulation of glucocorticoid on hypothalamus-pituitary-adrenal axis is closely related to Ca2 + and CaMPKⅡ.