氨氯地平对造影剂致人肾小管上皮细胞毒性的保护作用  

Protective effect of amlodipine on the cytotoxicity induced by contrast media in human kidney cells

作　　者：周晓蓉[1] 段绍斌[2] 彭佑铭[2] 刘伏友[2] 叶云[1] 刘瑞洪[1] 李桂源[3] 

机构地区：[1]中南大学湘雅二医院老年内科,长沙410011 [2]中南大学湘雅二医院肾内科,长沙410011 [3]中南大学肿瘤研究所,长沙410078

出　　处：《中南大学学报（医学版）》2007年第5期806-811,共6页Journal of Central South University ：Medical Science

基　　金：湖南省科委社会发展基金(1013-98)~~

摘　　要：目的:从细胞分子水平探讨氨氯地平对造影剂(泛影葡胺)致人肾小管上皮细胞毒性的保护作用及其机制。方法:选择人肾小管上皮细胞株(human kidney cell,HKC)为研究对象,实验分为4组,即模型组(泛影葡胺111g/L)、预防组(泛影葡胺111g/L+氨氯地平10-5mol/L)、氨氯地平对照组(氨氯地平10-5mol/L)和培养基对照组(单纯无血清DMEM-F12培养基),采用四甲基偶氮唑盐试验和细胞培养上清乳酸脱氢酶(lactate dehydrogenase,LDH)测定泛影葡胺对HKC细胞毒性,采用Ho-chest33258荧光染色和流式细胞仪DNA分析观察泛影葡胺诱导的肾小管上皮细胞凋亡。采用Western blot和免疫荧光法检测凋亡调控基因Bax的蛋白表达,采用荧光比色法检测半胱氨酸天冬氨酸蛋白酶-3(caspase-3)活性。结果:与模型组相比,预防组细胞活力明显增高,细胞培养上清中LDH浓度明显减低,细胞凋亡率显著降低,Bax蛋白表达下调,caspase-3活性降低。结论:氨氯地平可显著减轻泛影葡胺所致的肾小管上皮细胞损伤,对泛影葡胺诱导的肾小管上皮细胞凋亡有保护作用,其机制可能为通过Bax和caspase-3介导调控造影剂诱导的肾小管上皮细胞凋亡。Objective To explore the protective effect of amlodipine on the cytotoxicity in- duced by contrast media （meglumine diatrizoate ） in human kidney cells （HKC）. Methods An HKC line was used. The experiment was divided into 4 groups： a model group （diatrizoate 111 g/ L）, a prevention group （diatrizoate 111 g/L ＋ amlodipine 10^-5 mol/L） , an amlodipine control group （amlodipine 10^-5 mol/L） , and a culture medium control group （simple none blood serum DMEM-F12 medium）. Cytotoxicity induced by meglumine diatrizoate was analysed by methyl thia- zolyl tetrazolium （MTT） test, lactate dehydrogenase （LDH） assay, Hochest33258 fluorescence stained cytospins, and flow cytometric DNA analysis. The protein expression of Bax was determined by Western blot, and caspase-3 activity was examined by fluorometric method. Results In the pre-vention group, the cell viability increased significantly （ P 〈 0. 05 ） , LDH levels decreased （ P 〈 0.05 ） , and the apotosis was lower than that of the model group （ P 〈 0.05 ） . Bax protein expression and caspase 3 activity decreased （P 〈 0.05 ）. Conclusion Amlodipine can inhibit the HKC apoptosis and protect the renal ruble cell from injury induced by meglumine diatrizoate.

关 键 词：造影剂 人肾小管上皮细胞 细胞凋亡 钙拮抗剂 BAX CASPASE-3 

分 类 号：R96[医药卫生—药理学]