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【目的】研究铅对肾上腺皮质细胞氧化应激和线粒体功能的影响 ,为了解其肾上腺皮质毒作用机制提供依据。【方法】原代分离培养豚鼠肾上腺皮质细胞 ,以 0、6 2 5、12 5、2 5、5 0、10 0 μmol/L醋酸铅 (PbAc)处理细胞 ,观察PbAc诱导肾上腺皮质细胞活性氧 (ROS)产生和线粒体损伤作用。ROS检测采用荧光分光光度法 ,线粒体膜电位 (MMP)和细胞存活状态采用Rh12 3和PI双标记、流式细胞术检测 ,细胞ATP水平采用化学发光法测定。【结果】PbAc染毒后肾上腺皮质细胞ROS形成水平随剂量增加而增加 ,具有剂量效应关系 [^y=4 16 +10 2 1× 1g(x+1) ,P <0 0 1,R2 =0 6 41];线粒体膜电位呈剂量依赖性降低 ,Rh12 3的平均荧光强度 (MFI)在 6 2 5～ 10 0 μmol/L各剂量组依次为 1 0 1,0 94,0 96 ,0 95和 0 91,与对照组(1 35 )比较具有显著性差异 (P <0 0 1) ;染毒后细胞死亡率轻度增加 ,与对照组 (1 0 2 % )比较 ,5 0 μmol/L和 10 0 μmol/L的PbAc组分别为 3 16 %和 3 40 % ,差异具有显著性 (P <0 0 5 ) ;ATP水平降低与PbAc剂量之间存在剂量效应关系 [^y =2 12 96 5 7- 5 15 92 5× 1g(x +1) ,P <0 0 1,R2 =0 5 6 8],剂量和时间对ATP水平的影响呈协同抑制作用 (P <0 0 1)。【结论】线粒体氧化应激介导肾?
【Objective】 To investigate the effect of lead on oxidative stress and mitochondrial function in adrenocortical cells, and to provide basis for understanding its mechanism of adrenocortical toxicity. 【Methods】 Primary cultured rat adrenal cortical cells were cultured in the presence of 0, 62, 125, 25, 25, 50 and 10 0 μmol / L lead acetate (PbAc) to observe the effects of PbAc on ROS production in adrenocortical cells ) And mitochondrial damage. ROS detection using fluorescence spectrophotometry, mitochondrial membrane potential (MMP) and cell viability using Rh12 3 and PI double labeling, flow cytometry, cellular ATP levels were determined by chemiluminescence. 【Result】 The results showed that the level of ROS formation in adrenocortical cells increased with dose after exposure to PbAc, with a dose-response relationship [y = 4 16 +10 2 1 × 1g (x + 1), P 0 01, R2 0 The mitochondrial membrane potential decreased in a dose-dependent manner. The mean fluorescence intensity (MFI) of Rh12 3 in the range of 6 2 5 ~ 10 0 μmol / L was 1 0 1, 0 94, 0 96, 0 95 and There was a significant difference (P <0.01) between the control group and the control group (P <0.01). The cell death rate was slightly increased after the treatment. Compared with the control group (100% The difference was significant (P <0 05) between 10 0 μmol / L PbAc group and 3 16%, 3 40%, respectively. There was a dose-response relationship between decreased ATP level and PbAc dose The effect of dose and time on the ATP level was synergistically inhibited (P <0.01). 【Conclusion】 Mitochondrial oxidative stress mediates renal?