【摘要】 研究了 ATP 刺激的大鼠腹腔巨噬细胞[Ca²⁺]_i 升高与氧自由基（ROS）产生的关系和大黄酸抑制作用特征.提取大鼠腹腔巨噬细胞,利用 Ca²⁺探针 Fura-2检测单细胞胞内自由 Ca²⁺浓度([Ca²⁺]_i )变化,同时利用NBT 还原反应强度检测同一细胞 ROS 产生能力.结果发现无 ATP 刺激的巨噬细胞的 ROS 产生量较低;1mmol/L ATP 刺激巨噬细胞单细胞后诱发[Ca²⁺]_i 显著升高由胞内 Ca²⁺释放和胞外 Ca²⁺内流组成,同时 ROS产生增强2倍;胞外无 Ca²⁺条件下 ROS 产生随着[Ca²⁺]_i 下降而减少.10^-5和10^-4mol/L 浓度大黄酸对1mmol/L ATP 刺激巨噬细胞单细胞的[Ca²⁺]_i 升高和 ROS 产生有剂量依赖性的抑制作用;多细胞的统计分析表明10^-5和10^-4mol/L 浓度大黄酸分别抑制了[Ca²⁺]_i 峰值的49%和84%,同时抑制了 ROS 产生的59%和81%.因此认为 ATP 刺激大鼠腹腔巨噬细胞诱发的[Ca²⁺]_i 升高介导了 ROS 的产生,大黄酸剂量依赖性的抑制 ATP 刺激的细胞[Ca²⁺]_i 升高和 ROS 产生能力,并提示大黄酸抑制细胞[Ca²⁺]_i 升高是其抑制 ROS 产生的重要机制.更多还原

【Abstract】 In order to investigate the relation between increase in [Ca²⁺]_i and ROS generat- ing activity stimulated by ATP,and the effect of rhein on increase in [Ca²⁺]_i and ROS generat- ing activity in peritoneal macrophages’（PM）.The [Ca²⁺]_i of single cell was measured by fluo- rescent microscopic imaging system after loading with fluorescent probe fura-2/AM,ROS gen- erating activity was determined by modified NBT test in same cell.Results show that after treatment of 1 mmol/L ATP,the [Ca²⁺]_i was increased by Ca²⁺ release from intracellular stores and Ca²⁺ influx,and the intensity of NBT reduction test was increased two times in same cell.When Ca²⁺-free medium,the ROS generating activity was decreased with [Ca²⁺]_i was de- clined.Rhein dose-dependently inhabited [Ca²⁺]_i and ROS generating activity at 10^-5and 10^-4 mol/L.Multicellular results show 10^-5 and 10^-4 mol/L rhein inhibited 49% and 84% of the peak of [Ca²⁺]_i and 59% and 81% of ROS generating activity respectively.We can come to the conclusion the increase in[Ca²⁺]_i induced ROS generating activity in ATP stimulated PM. rhein dose-dependently inhibites increase in [Ca²⁺]_i and ROS generating activity,rhein inhib- ites ROS generating activity via a pathway dependent on reducing [Ca²⁺]_i.更多还原