摘要
体外培养的HK-2细胞分为正常对照组（NG组,5.5mmol/LD-葡萄糖）、高糖处理组（HG组,30mmol/LD-葡萄糖）、高渗对照组（MG组,5.5mmol/LD-葡萄糖＋24.5mmol/LD-甘露醇）、高糖培养液中又加入不同浓度的1,25-二羟维生素D3[1,25-（OH）2D3]组（V1～V3组）、N-乙酰半胱氨酸药效对照组（NAC组,30mmol/LD-葡萄糖＋1.0mmol/LN-乙酰半胱氨酸）和无水乙醇溶剂对照组（SG组,30mmol/LD-葡萄糖＋6.86×10-2mol/L乙醇）。检测细胞活性氧簇荧光强度、线粒体膜电位、解耦联蛋白2（UCP2）mRNA、蛋白表达、总超氧化物歧化酶（SOD）活力和丙二醛水平。结果HG组细胞的线粒体膜电位明显低于NG组（P〈0.01）,1,25-（OH）2D3处理后细胞的线粒体膜电位与HG组比较显著下降（均P〈0.01）;HG组总SOD活力显著低于NG组（P〈0.01）,丙二醛含量明显高于NG组（P〈0.01）,而1,25-（OH）2D3处理后细胞总SOD活力较HG组显著增高（P〈0.05）,丙二醛水平显著低于HG组（P〈0.01）;HG组UCP2mRNA与蛋白表达显著高于NG组（P〈0.05）,而1,25-（OH）2D3处理后细胞UCP2的表达与HG组比较显著降低（P〈0.01）。结果表明高糖可诱导体外培养的HK-2细胞氧化应激损伤;1,25-（OH）2D3可能通过降低线粒体膜电位、活性氧簇生成及调节胞内UCP2表达抑制高糖诱导的氧化应激反应。
The HK-2 cells with different culture media were divided into normal glucose group（NG group, 5.5 mmol/L D-glucose）; high glucose group（HG group, 30 mmol/L D-glucose）; mannitol group（MG group, 5.5 mmol/L D-glucose＋24.5 mmol/L mannitol）; 1,25-dihydroxyvitamin D3[1,25-（OH）2D3] groups（V1-V3 group）which were exposed to medium containing 30 mmol/L D-glucose and different concentrations of 1,25-（OH）2D3; Nethyl-cysteim control group（NAC group, 30 mmol/L D-glucose ＋1.0 mmol/L N-Nethyl-cysteim）; and ethanol control group（SG group, 30 mmol/L D-glucose＋6.86×10-2 mol/L ethanol）. The level of intracellular reactive oxygen species, mitochondrial membrane potential, activity of total-superoxide dismutase（T-SOD）, level of malondialdehyde, expression of UCP2 mRNA and protein in HK-2 cells were detected. Compared with NG group, the mitochondrial membrane potential significantly decreased in HG group（P〈0.01）, and the mitochondrial membrane potential in V group was lower than that in HG group（P〈0.01）. The activity of T-SOD in HG group was significantly lower than that in NG group（P〈0.01）, while its level of malondialdehyde was significantly higher than that in NG group（P〈0.01）. Compared with HG group, the activity of T-SOD in V groups was significantly increased（P〈0.05）and the level of malondialdehyde in these groups significantly decreased（P〈0.01）. The mRNA expression of UCP2 in HG group was increased significantly in comparison with NG group（P〈0.05）and the expression in V groups was significantly decreased in comparison with HG group（ P〈0.01）. The results suggest that 1,25-（OH）2D3 could reduce the mitochondrial membrane potential, the production of reactive oxygen species, and regulate the expression of UCP2 in order to suppress the oxidative stress induced by high glucose.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2013年第10期849-852,共4页
Chinese Journal of Endocrinology and Metabolism
关键词
糖尿病肾病
1
25-二羟维生素D3
HK-2细胞
解耦联蛋白2
氧化应激
Diabetic nephropathy
1,25-dihydroxyvitamin D3
Human renal tubular epithelial cells
Uncoupling protein 2
Oxidative stress
