雷洛昔芬对氯化镉染毒肾小管上皮细胞的影响  

Effect of Raloxifene on Cadmium Chloride-Contaminated Renal Tubular Epithelial Cells

作　　者：肖敏[1] 尹友生[1] 冷斌[1] 李清初[1] 李康慧[1] 韦家智[1] 

机构地区：[1]桂林医学院附属医院肾内科,桂林541000

出　　处：《医药导报》2016年第2期120-124,共5页Herald of Medicine

基　　金：广西医疗卫生重点科研课题(重2012011)

摘　　要：目的观察雷洛昔芬对氯化镉染毒肾小管上皮细胞中雌激素受体α(ERα)、ERβ和金属硫蛋白-1a(MT-1a)基因表达的影响,并探讨其机制。方法选取新生Sprague Dawley(SD)大鼠的肾脏组织进行细胞培养,采用Percoll法分离纯化肾小管后原代培养。选取第2代细胞进行实验。实验分为空白对照组、氯化镉组、雷洛昔芬组。4 h后采用噻唑蓝(MTT)法检测细胞活力,24 h后应用反转录聚合酶链反应(RT-PCR)及免疫组化检测ERα、ERβ及MT-1a mRNA及蛋白水平。结果与空白对照组比较,氯化镉组大量细胞的形态发生改变,细胞大量死亡,存活率为55.3%,同时ERβ,MT-1a mRNA及蛋白表达水平升高(P<0.05);与氯化镉组比较,雷洛昔芬组仅少量细胞形态发生改变,存活率88.6%,ERβ,MT-1a mRNA及蛋白表达水平下降(P<0.05)。而ERα表达在3组间变化不明显,差异无统计学意义(P>0.05)。结论雷洛昔芬可竞争性与ERβ结合,下调MT-1a的表达,减少Cd-MT复合物的形成,而降低镉对肾小管上皮细胞的损伤。Objective To investigate the effect of raloxifene on ERα,ERβ and MT-1a expression in renal tubular epithelial cells contaminated with cadmium chloride,for exploring action mechanism. Methods Renal tubule cells were isolated from kidneys of new born SD rats and purified by Percoll. The cells of the second generation were selected for experiment.The cells were divided into 3 groups： blank control group,cadmium chloride group and raloxifene group. After 4 h,cell viability was detected by MTT,and after 24 h,mRNA and protein expression levels of ERα,ERβ and MT-1a in renal tissues were determined by RT-PCR and immunohistochemistry technology,respectively. Results Compared with the blank control group,morphology of plenty cells were changed in cadmium chloride group,a number of cells were dead,and the survival rate was only55. 3%; at the same time,mRNA and protein expression levels of ERβ and MT-1a significantly increased（ P〈0. 05）. Compared with cadmium chloride group,only a small part of cells underwent morphological changes,and the survival rate was 88. 6% in raloxifene group,and the mRNA and protein expression levels of ER β and MT-1a were significantly decreased（ P〈0. 05）. The expression of ERα showed no significant difference among the three groups（ P 〈0. 05）. Conclusion Raloxifene can bond with ERβ competively,down-regulate MT-1a and decrease Cd-MT synthesis,so as to reduce cadmium-induced injury of renal tubular epithelial cells.

关 键 词：雷洛昔芬 镉 雌激素受体 金属硫蛋白 上皮细胞 肾小管 

分 类 号：R971.1[医药卫生—药品] R965[医药卫生—药学]