去甲肾上腺素引起C6胶质瘤细胞中钙的动员

目的:研究去甲肾上腺素引起大鼠C6神经胶质瘤细胞中钙离子浓度([Ca~(2 )])增加的机理.方法:以荧光染料fura-2为指示剂,采用双波长荧光比值成像 的方法测定细胞中钙离子浓度.结果:通过激活细胞上的α_1肾上腺素能受体,去甲肾上腺素剂量依赖地使C6细胞中钙离子浓度增加.这种反应不依赖细胞外钙,且不受百日咳毒素(PTX)处理的影响.将细胞与磷酯酶C(PLC)抑制剂U73122或内质网Ca~(2 )-ATP酶抑制剂thapsigargin预孵育,去甲肾上腺素引起的胞内钙反应则消失;蛋白激酶C激动剂佛波醉酯(PMA)预处理细胞可以使去甲肾上腺素引起的胞内钙离子浓度升高幅度降低,而佛波醇酯的效应能够被蛋白激酶C抑制剂Ro31-8220或GF-109203X完全阻断.但是,改变胞内蛋白激酶A活性的药物对去甲肾上腺素的作用没有影响.结论:通过激活胞内的磷酯酶C,去甲肾上腺素使C6细胞的胞内钙库释放钙离子.去甲肾上腺素引起的细胞内钙离子浓度增加受蛋白激酶C的负性调节.

Norepinephrine-induced calcium mobilization in C6 glioma cells

AIM: To investigate the mechanism underlying the norepinephrine-induced elevation in intracellular calcium concentration ([Ca2+]i) in C6 glioma cells. METHODS: Measurement of [Ca2+]i was carried out using the dual-wavelength fluorescence method with fura-2 as the indicator. RESULTS: Norepinephrine was found to induce concentration-dependent increases in [Ca2+ ]i through alpha1-adrenoreceptors. The [Ca2+]i elevations were extracellular-calcium independent and not influenced by the treatment of pertussis toxin. Pretreatments with either U73122 or thapsigargin abolished the subsequent cellular calcium responses to norepinephrine. Preincubation with phorbol 12-myristate 13-acetate (PMA) significantly reduce d the [Ca2+]i elevations, while protein kinase C inhibitors Ro31-8220 or GF-109203X completely blocked the inhibitory action of PMA. However, drugs either activating or inhibiting the function of protein kinase A had no effect on the [Ca2+]i elevations. CONCLUSION: Norepinephrine induces calcium mobilization from internal stores by activation of phospholipase C in C6 cells. The [Ca2+]i elevation is negatively regulated by the activation of protein kinase C.