摘要
研究了 ATP 刺激的大鼠腹腔巨噬细胞[Ca^(2+)]_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 Ca^(2+)探针 Fura-2检测单细胞胞内自由 Ca^(2+)浓度([Ca^(2+)]_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[Ca^(2+)]_i 显著升高由胞内 Ca^(2+)释放和胞外 Ca^(2+)内流组成,同时 ROS产生增强2倍;胞外无 Ca^(2+)条件下 ROS 产生随着[Ca^(2+)]_i 下降而减少.10^(-5)和10^(-4)mol/L 浓度大黄酸对1mmol/L ATP 刺激巨噬细胞单细胞的[Ca^(2+)]_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^(-4)mol/L 浓度大黄酸分别抑制了[Ca^(2+)]_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[Ca^(2+)]_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[Ca^(2+)]_i 升高和 ROS 产生能力,并提示大黄酸抑制细胞[Ca^(2+)]_i 升高是其抑制 ROS 产生的重要机制.
In order to investigate the relation between increase in [Ca^2＋]i and ROS generating activity stimulated by ATP, and the effect of rhein on increase in [Ca^2＋]i and ROS generating activity in peritoneal macrophages＇ （PM）. The [Ca^2＋]i of single cell was measured by fluorescent microscopic imaging system after loading with fluorescent probe fura-2/AM, ROS generating activity was determined by modified NBT test in same cell. Results show that after treatment of 1 mmol/L ATP , the [-Caz＋ ]i was increased by Ca^2＋ release from intracellular stores and Ca^2＋ influx, and the intensity of NBT reduction test was increased two times in same cell. When Ca^2＋ -free medium, the ROS generating activity was decreased with [Ca^2＋]i was declined . Rhein dose-dependently inhabited [Ca^2＋]i and ROS generating activity at 10^-5 and 10^-4 mol/L. Multicellular results show 10^-5 and 10^-4 mol/L rhein inhibited 49 % and 84 % of the peak of [Ca^2＋]i and 59 % and 81 %of ROS generating activity respectively. We can come to the conclusion the increase in [Ca^2＋]i induced ROS generating activity in ATP stimulated PM. rhein dose-dependently inhibites increase in [Ca^2＋]i and ROS generating activity, rhein inhib- ites ROS generating activity via a pathway dependent on reducing [Ca^2＋]i.
出处
《南开大学学报（自然科学版）》
CAS
CSCD
北大核心
2007年第5期96-102,共7页
Acta Scientiarum Naturalium Universitatis Nankaiensis
基金
天津市科技攻资助项目(983113411)
