Yuan Lin, Han Guangliang
School of Public Health, Xinxiang Medical University, Xinxiang 453003, China.
Wei Sheng Yan Jiu. 2018 Mar;47(2):290-295.
To observe the effects of cadmium chloride on mitochondrial function and the expression of PGC-1α in HK-2 cells, and explore the possible role of cadmium chloride in the process of mitochondrial oxidative damage of HK-2 cells.
HK-2 cells were exposed to 0, 10, 20, 30, 40, 50 and 60 μmol/L of CdCl_2 respectively for 24 h in vitro. MTT assay was used to observe the viability of HK-2 cells. The mitochondrial ROS was detected by Mito SOXTMRed staining. Mitochondrial membrane potential was measured by JC-1 staining flow cytometry. The activity of respiratory chain complex III was determined by Multi-functional microplate reader and PGC-1α expression was detected by Western blot analysis.
Compared with control, CdCl_2 treatment at 20 and 60 μmol/L for 24 h the survival rate of HK-2 cells was decreased from( 77. 60 ± 0. 82) % ( 41. 97 ± 1. 22) %( P < 0. 01). The activity ofrespiratory chain complex III was decreased in a concentration-dependent manner( P <0. 05), while the mitochondrial ROS production was increased at the same time( P <0. 05), the JC-1 monomers positive ratio by the percentage of the treated cells was 1. 51, 1. 58, 1. 71, 2. 41 3. 47 times higher than untreated control( P < 0. 01). In addition, the expression of PGC-1α was decreased at 24 h( P < 0. 05).
Cadmium chloride may inhibit the expression of PGC-1α and the activity of respiratory chain complex III, and by inducing the mitochondrial ROS production and reducing mitochondrial membrane potential, leading to the damage of HK-2 cells.
观察氯化镉对HK-2细胞线粒体功能及PGC-1α表达的影响，探讨氯化镉在HK-2细胞线粒体氧化损伤过程中的可能作用。
将HK-2细胞分别体外暴露于0、10、20、30、40、50和60 μmol/L的CdCl₂ 24小时。采用MTT法观察HK-2细胞的活力。通过Mito SOXTMRed染色检测线粒体活性氧。采用JC-1染色流式细胞术检测线粒体膜电位。用多功能酶标仪测定呼吸链复合体III的活性，通过蛋白质免疫印迹分析检测PGC-1α的表达。
与对照组相比，20和60 μmol/L的CdCl₂处理24小时后，HK-2细胞的存活率从(77.60±0.82)%降至(41.97±1.22)%(P<0.01)。呼吸链复合体III的活性呈浓度依赖性降低(P<0.05)，而线粒体活性氧的产生同时增加(P<0.05)，处理组细胞的JC-1单体阳性率是未处理对照组的1.51、1.58、1.71、2.41、3.47倍(P<0.01)。此外，24小时时PGC-1α的表达降低(P<0.05)。
氯化镉可能抑制PGC-1α的表达和呼吸链复合体III的活性，并通过诱导线粒体活性氧的产生和降低线粒体膜电位，导致HK-2细胞损伤。
