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本研究旨在阐明过氧化氢（Ｈ＿２Ｏ＿２）和膜钠钙交换蛋白相互作用对胞浆钙〔Ｃａ￣２＋〕＿ｉ的调控。在稳定表达钠钙交换蛋白ＣＫ１．４细胞上，用￣（４５）Ｃａ同位素液闪计数法测定钠钙交换蛋白的活性；用ｆｕｒａ─２荧光探针和３４０／３８０ｎｍ双兴奋波长荧光影像技术测定钙释放和〔Ｃａ￣２＋〕ｉ。两因素两水平和三因素两水平正交分析表明１０ｍｍｏｌ／ＬＨ＿２Ｏ＿２与１５０ｍｍｏｌ／Ｌ细胞外钠〔Ｎａ￣＋〕ｏ、１ｍｍｏｌ／Ｌ细胞外钙〔Ｃａ￣２＋〕。相互作用或１０ｍｍｏｌ／ＬＨ＿２Ｏ＿２分别与１５０ｍｍｏｌ／Ｌ〔Ｎａ￣＋〕。或１ｍｍｏｌ／Ｌ〔Ｃａ￣２＋〕。相互协同，均增加钠钙交换蛋白的活性，抑制钙内流。１４０ｍｍｏｌ／Ｌ〔Ｎａ￣＋〕。激活钠钙交换蛋白，排出细胞内钙离子，降低〔Ｃａ￣２＋〕＿ｉ。当〔Ｎａ￣＋〕。递减至０ｍｍｏｌ／Ｌ时，１０ｍｍｏｌ／ＬＨ＿２Ｏ＿２直接抑制钠钙交换蛋白的活性，增加钙释放和升高〔Ｃａ￣２＋〕＿ｉ。在不同〔Ｎａ￣＋〕。梯度中，１０ｍｍｏｌ／ＬＨ＿２Ｏ＿２对膜钠钙交换活动和〔Ｃａ￣２＋〕＿ｉ起双重调节作用，即抑制或增加钙内流和〔Ｃａ￣２＋〕＿ｉ。１０ｍｍｏｌ／ＬＨ＿２Ｏ＿２与膜钠钙交换蛋白和〔Ｃａ￣２＋〕?
The purpose of this study was to elucidate the regulation of cytoplasmic calcium [Ca2 +] i by the interaction of hydrogen peroxide (H2O2) and membrane sodium-calcium exchange protein. The activity of sodium-calcium exchanger was determined by ~ (45) Ca isotope liquid scintillation counting assay on stable expression of sodium-calcium exchange protein CK1.4. The fluorescence intensity of 340 ± 380 nm Calcium release and 〔Ca ~ 2 +〕 i. Orthogonal analysis of two factors and two levels and three factors and two levels showed that 10mmol / L H2O2 and 150mmol / L extracellular Na [Na ~ +] o, 1mmol / L extracellular calcium [Ca ~ 2 +]. Interaction or 10mmol / LH_2O_2 with 150mmol / L 〔Na ~ +〕. Or 1 mmol / L [Ca ~ 2 +]. Mutual synergy, increase sodium-calcium exchange protein activity, inhibit calcium influx. 140 mmol / L [Na ~ +]. Activate sodium-calcium exchange protein, excrete intracellular calcium, reduce [Ca ~ 2 +] _i. When 〔Na ~ +〕. Decreased to 0mmol / L, 10mmol / LH_2O_2 directly inhibit the activity of sodium-calcium exchanger, increase calcium release and increase [Ca ~ 2 +] _i. In different 〔Na ~ +〕. In the gradient, 10 mmol / LH 2 O 2 plays a dual regulatory role for membrane sodium-calcium exchange activity and [Ca 2+] i, which inhibits or increases calcium influx and [Ca 2+] i. 10mmol / LH_2O_2 and membrane sodium-calcium exchange protein and 〔Ca ~ 2 +〕?