白藜芦醇对碘普罗胺诱导人肾小管上皮HK-2细胞氧化应激及Klotho蛋白表达的影响  

The effect of resveratrol on oxidative stress and Klotho expression in iopromide-induced human renal tubular epithelial HK-2 cells

作　　者：张滨[1] 钟小华[1] 廖丽贞 

机构地区：[1]广东药科大学附属第一医院康复医学科,广东广州510080 [2]广东药科大学健康学院,广东广州510006

出　　处：《广东医学》2018年第4期501-505,共5页Guangdong Medical Journal

基　　金：广东省中医药局建设中医药强省立项资助科研课题(编号:20152155)

摘　　要：目的探讨白藜芦醇对碘普罗胺诱导的人肾小管上皮HK-2细胞氧化应激及Klotho蛋白表达的影响。方法应用不同剂量(137.5、275.0和550.0 mg I/mg)的碘普罗胺处理人肾小管上皮HK-2细胞12 h建立离体的对比剂肾病模型,以MTT比色法检测细胞存活率、试剂盒法检测乳酸脱氢酶酶(LDH)活力以及流式细胞术检测细胞凋亡百分率。加入不同剂量(1、10和100μmol/L)的白藜芦醇或N-乙酰半胱氨(NAC)处理,检测碘普罗胺诱导的HK-2细胞存活率、LDH和细胞凋亡百分率的变化,同时使用CMH2DCFDA荧光法检测细胞内活性氧簇(ROS)荧光强度,采用Western bolt法检测各组细胞的Klotho蛋白表达情况。结果碘普罗胺处理人肾小管上皮HK-2细胞12 h后,HK-2细胞的存活率呈剂量依赖性下降,LDH和凋亡率均呈剂量依赖性地升高;加入不同浓度白藜芦醇处理后,白藜芦醇能呈剂量依赖性地抑制碘普罗胺所致的细胞存活率下降、LDH和凋亡率的升高,效果与NAC类似。此外,与对照组比较,碘普罗胺组的ROS荧光染色增加。加入白藜芦醇或NAC后,均能抑制由碘普罗胺所致的ROS荧光染色增加。在Klotho蛋白表达方面,与对照组比较,碘普罗胺组的Klotho蛋白表达下调,加入白藜芦醇后,能有效抑制由碘普罗胺所致的Klotho蛋白表达下调,而加入NAC后,并不能有效抑制由碘普罗胺所致的Klotho蛋白表达下调,其Klotho蛋白表达与单纯碘普罗胺组相当。结论白藜芦醇具有抑制碘普罗胺所致的HK-2细胞毒性作用,效果与NAC相近,其机制可能与白藜芦醇上调Klotho蛋白从而降低碘普罗胺所致的HK-2细胞氧化应激水平有关。Objective To investigate the effect of resveratrol on oxidative stress and Klotho expression in iopro- mide - induced human renal tubular epithelial HK - 2 cells. Methods Different doses of iopromide （ 137.5 mg/mg, 275.0 mgI/mg and 550.0 mgI/mg） were added in human renal tubular epithelial HK - 2 cells for 12 h to mimic contrast -induced nephropathy in vitro. MTT assay was used to measure the cell viability. Kit assay was used to measure lactate dehydrogenase enzyme （LDH） vitality and flow cytometry was used to measure percentage of apoptosis. Different doses （1 μmol/L, 10 μmol/L and 100 μmol/L） of resveratrol or N -acetylcysteine （NAC） was added, and the cell viability, LDH vitality and percentage of apoptosis were detected. CM - H2DCFDA assay was used to measure cell fluorescence in- tensity of reactive oxygen species （IOS）. Western bolt assay was used to measure the expression of Klotho protein. Re- sults Iopromide treatment for 12 h in human renal tubular epithelial HK -2 cells caused cell viability decrease, and ele- vated LDH vitality and apoptosis in a dose - dependent manner. When different doses of resveratrol was added, the above cell changes were inhibited in a dose - dependent manner. The effect of resveratrol was similar to the NAC. In addition, iopromide treatment caused ROS fluorescence intensity upregulation, which was inhibited by both resveratrol and NAC. The Klotho protein expression was reduced in the iopromide group when compared with the control group, which was inhib- ited by resveratrol but not NAC. Conclusion Resveratrol alleviates iopromide - induced toxic effects on HK - 2 cells as the same as NAC. It may be related to its effects on upregulation of Klotho protein and downregulation of oxidative stress inHK - 2 cells.

关 键 词：白藜芦醇 碘普罗胺 人肾小管上皮HK^2细胞 KLOTHO蛋白 氧化应激 凋亡 

分 类 号：R692[医药卫生—泌尿科学]