摘要
目的探讨氯化镉对人肾小管上皮细胞(HK-2细胞)肾损伤分子-1(KIM-1)表达的影响。方法取对数生长期的HK-2细胞分为对照组和5个氯化镉处理组,分别以终浓度为0.0、5.0、10.0、20.0、50.0、100.0μmol/L(以磷酸盐缓冲液为溶剂)的氯化镉处理。培养24 h后进行细胞病理学观察,采用噻唑蓝实验检测HK-2细胞存活率,分别以逆转录聚合酶链反应法、免疫印迹法检测KIM-1的mRNA和蛋白表达水平。结果对照组和5.0、10.0μmol/L组HK-2细胞形态无明显变化;20.0和50.0μmol/L组HK-2细胞出现不同程度的肿胀或空泡样改变;100.0μmol/L组HK-2细胞大量死亡。20.0、50.0和100.0μmol/L组HK-2细胞存活率分别低于对照组和5.0、10.0μmol/L组(P<0.05);20.0、50.0和100.0μmol/L组细胞存活率两两比较,差异均有统计学意义(P<0.05)。20.0和50.0μmol/L组KIM-1的mRNA和蛋白表达水平分别高于对照组和5.0、10.0μmol/L组(P<0.05);50.0μmol/L组细胞KIM-1的mRNA和蛋白表达水平均高于20.0μmol/L组(P<0.05)。结论一定水平范围的氯化镉可上调HK-2细胞KIM-1的mRNA和蛋白表达;KIM-1可能是镉致肾小管损伤效应的生物标志物之一。
Objective To investigate the effect of cadmium chloride on the expression of kidney injury molecule-1（ KIM-1）in human renal tubular epithelial cells（ HK-2 cells）. Methods HK-2 cells at logarithmic phase were divided into a control group and 5 treatment groups that were treated with 5. 0,10. 0,20. 0,50. 0 and 100. 0 μmol / L of cadmium chloride dissolved in phosphate buffer solution. Cell pathology observation was carried out after 24 hours of cultivation. The methyl thiazolyl tetrazolium assay was used to calculate the survival rate of HK-2 cells. The expression of KIM-1 mRNA and protein were detected by the reverse transcription-polymerase chain reaction and Western blotting analysis respectively.Results There were no cellular morphologic change in HK-2 cells in the control group,the 5. 0 and 10. 0 μmol / L groups;the HK-2 cells showed different degree of swellings or vacuoles in the 20. 0 and 50. 0 μmol / L groups; a large number of cells were found dead in the 100. 0 μmol / L group. The cell survival rates of HK-2 cells in the 20. 0,50. 0 and 100. 0μmol /L groups were lower than those of control group,the 5. 0 and 10. 0 μmol /L groups（ P〈0. 05）. The pairwise comparison among survival rates of the 20. 0,50. 0 and 100. 0 μmol / L groups showed significant difference（ P〈0. 05）.The expression levels of KIM-1 mRNA and protein in the 20. 0 and 50. 0 μmol / L groups were higher than those of control group,the 5. 0 and 10. 0 μmol / L groups（ P〈0. 05）. The levels of KIM-1 mRNA and protein in the 50. 0 μmol / L group were higher than those of the 20. 0 μmol / L group（ P〈0. 05）. Conclusion Cadmium chloride at certain concentration can increase the expression of KIM-1 mRNA and protein in HK-2 cells. Therefore,the expression of KIM-1 could be used as one of the effect biomarkers for cadmium induced kidney tubule injury.
出处
《中国职业医学》
CAS
北大核心
2016年第1期37-41,共5页
China Occupational Medicine
基金
国家职业病临床重点专科(WY2011873)
深圳市科技创新计划项目(JCYJ20130401092802783)
深圳市重大职业病诊疗技术重点实验室建设项目(CXB201111250112A)
