雷洛昔芬对镉染毒后肾小管上皮细胞存活情况及1-α羟化酶表达的影响

发布时间：2018-05-10 15:30

本文选题：肾小管上皮细胞 + 雷洛昔芬　；参考：《山东医药》2016年30期

【摘要】：目的观察应用雷洛昔芬对镉染毒的肾小管上皮细胞存活情况及1-α羟化酶表达的影响,探讨雷洛昔芬治疗镉中毒导致肾毒性的机制。方法分离纯化SD大鼠肾小管上皮细胞原代细胞。分别用浓度为2.5、5.0、10.0、20.0、40.0μmol/L的氯化镉对肾小管上皮细胞染毒24 h,采用MTT法测算肾小管上皮细胞的相对存活率。选取20.0μmol/L的氯化镉剂量作用于肾小管上皮细胞,分别用0.05、0.10、0.50、1.00μmol/L的雷洛昔芬干预24h,采用MTT法测算肾小管上皮细胞的相对存活率。将肾小管上皮细胞分为空白对照组、氯化镉组、雷洛昔芬组,空白对照组培养于DMEM培养基,氯化镉组以20.0μmol/L的氯化镉染毒肾小管上皮细胞,雷洛昔芬组以20.0μmol/L的氯化镉染毒后给予雷洛昔芬0.10μmol/L干预,均干预24 h,在显微镜下观察细胞形态,采用RT-PCR法检测肾小管上皮细胞1-α羟化酶mRNA表达。结果 2.5、5.0、10.0、20.0、40.0μmol/L的氯化镉干预24 h后,肾小管上皮细胞的相对存活率均降低(P均0.01),且氯化镉浓度较高时细胞相对存活率较低。对镉染毒后的肾小管上皮细胞以0.05、0.10、0.50、1.00μmol/L的雷洛昔芬干预24 h后,肾小管上皮细胞的相对存活率均高于镉染毒细胞(P均0.01)。空白对照组肾小管上皮细胞形态正常,氯化镉组肾小管上皮细胞形态异常、大量细胞死亡,雷洛昔芬组细胞形态异常较氯化镉组明显减轻。氯化镉组肾小管上皮细胞1-α羟化酶mRNA相对表达量低于空白对照组,雷洛昔芬组1-α羟化酶mRNA相对表达量高于氯化镉组(P均0.05)。结论雷洛昔芬对氯化镉染毒的肾小管上皮细胞具有保护作用,可上调镉染毒后肾小管上皮细胞1-α羟化酶mRNA的表达。
[Abstract]:Objective to observe the effects of raloxifene on the survival of cadmium-exposed renal tubular epithelial cells and the expression of 1- 伪 -hydroxylase, and to explore the mechanism of raloxifene in the treatment of renal toxicity induced by cadmium poisoning. Methods SD rat renal tubular epithelial cells were isolated and purified. The relative survival rate of renal tubular epithelial cells was measured by MTT assay after exposure to cadmium chloride of 2.5V 5.0 渭 mol/L (10.0 渭 mol/L) for 24 hours. The renal tubular epithelial cells were treated with 20 渭 mol/L cadmium chloride for 24 h. The relative survival rate of renal tubular epithelial cells was measured by MTT method. Renal tubular epithelial cells were divided into blank control group, cadmium chloride group, raloxifene group, blank control group, cultured in DMEM medium. Cadmium chloride group was exposed to 20 渭 mol/L cadmium chloride on renal tubular epithelial cells. Raloxifene group was treated with 20 渭 mol/L cadmium chloride for 24 h. The cell morphology was observed under microscope and the expression of 1- 伪 hydroxylase mRNA in renal tubular epithelial cells was detected by RT-PCR method. Results the relative survival rate of renal tubule epithelial cells was decreased after 24 hours of cadmium chloride treatment with 2.55.010.0 渭 mol/L (20.0 渭 mol/L), and the relative survival rate of renal tubular epithelial cells was lower when cadmium chloride concentration was higher than that of the control group (P < 0.05), and the relative survival rate of renal tubular epithelial cells was lower when the concentration of cadmium chloride was higher. The relative survival rate of renal tubular epithelial cells exposed to cadmium was higher than that of cadmium exposed cells (P < 0.01). The morphology of renal tubular epithelial cells in the blank control group was normal, and that in the cadmium chloride group was abnormal and a large number of cells died. The abnormal morphology of the renal tubular epithelial cells in the raloxifen group was significantly less than that in the cadmium chloride group. The relative expression of 1- 伪 -hydroxylase mRNA in renal tubular epithelial cells in cadmium chloride group was lower than that in control group, and the relative expression of 1- 伪 -hydroxylase mRNA in raloxifene group was higher than that in cadmium chloride group (P < 0.05). Conclusion Riloxifene has protective effect on renal tubular epithelial cells exposed to cadmium chloride and can up-regulate the expression of 1- 伪 -hydroxylase mRNA in renal tubular epithelial cells after cadmium exposure.
【作者单位】：桂林医学院附属医院;
【基金】：广西医疗卫生重点科研课题(重2012011)
【分类号】：R114;R692

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