达格列净对高糖诱导的人近端肾小管细胞损伤的保护作用  

Protective effect of dapagliflozin on high glucose-induced injury in human proximal renal tubular cells

作　　者：孙凌云[1] 蔡佳盈[1] 沈世忠[1] 王晓松[1] 吴家祥 SUN Ling-yun;CAI Jia-ying;SHEN Shi-zhong;WANG Xiao-song;WU Jia-xiang(Department of Nephrology,Quanzhou First Hospital,Fujian Medical University,Quanzhou 362000,Fujian Province,China;School of Clinical Medicine,Quanzhou Medical College,Quanzhou 362000,Fujian Province,China)

机构地区：[1]福建医科大学附属泉州第一医院肾内科,福建泉州362000 [2]福建省泉州医学高等专科学校临床医学院,福建泉州362000

出　　处：《中国临床药理学杂志》2022年第8期777-781,共5页The Chinese Journal of Clinical Pharmacology

基　　金：福建省教育厅中青年教师教育科研基金资助项目(科技类,JAT171166);泉州医学高等专科学校重点科技基金资助项目(XJK1602A)。

摘　　要：目的研究达格列净对高糖诱导的人近端肾小管上皮细胞(HK-2)损伤的保护作用。方法将HK-2细胞随机分为空白组、对照组、模型组和实验组。空白组不进行任何干预;对照组加入5μmol·L^(-1)达格列净;模型组加入25 mmol·L^(-1)葡萄糖;实验组加入25 mmol·L^(-1)葡萄糖和5μmol·L^(-1)达格列净,各组细胞培养24 h后进行后续检测。用流式细胞术检测各组细胞凋亡情况,用噻唑蓝(MTT)法检测各组细胞增殖情况,用酶生化法和荧光显微镜法检测各组细胞内丙二醛(MDA)和活性氧(ROS)水平,用酶联免疫吸附法检测各组细胞内超氧化物歧化酶(SOD)和转化生长因子-β1(TGF-β1)、肝细胞生长因子(HGF)水平。结果空白组、对照组、模型组和实验组HK-2细胞的凋亡率分别为(4.73±0.46)%,(4.46±0.61)%,(35.42±2.37)%和(19.55±1.87)%;细胞增殖光密度值分别为0.52±0.09,0.49±0.07,0.13±0.03和0.29±0.05;细胞内MDA水平分别为(3.59±0.37),(4.13±0.52),(9.87±2.31)和(5.46±0.94)μmol·L^(-1);SOD水平分别为(3.75±0.31),(3.46±0.27),(2.03±0.16)和(3.12±0.22)kU·g Hb^(-1);ROS水平分别为(3.12±0.55)%,(3.24±0.53)%,(14.67±2.64)%和(8.37±1.43)%;细胞内TGF-β1水平分别为(446.31±55.42),(435.68±53.22),(1667.49±104.52)和(1235.74±95.35)pg·mg^(-1);HGF水平分别为(949.75±84.35),(964.84±68.26),(635.34±66.81)和(808.12±77.10)pg·mg^(-1)。以上指标,模型组与空白组比较,差异均有统计学意义(均P<0.05);实验组与模型组比较,差异均有统计学意义(均P<0.05)。结论达格列净能有效抑制高糖诱导的HK-2细胞凋亡,促进HK-2细胞,还能有效降低细胞内氧化应激和肾损伤标志物的水平,可能有助于糖尿病肾病的治疗和预防。Objective To study the protective effect of dapagliflozin on human proximal renal tubular epithelial cell(HK-2)injury induced by high glucose.Methods HK-2 cells were randomly divided into blank group,control group,model group and experimental group.Blank group were not given any intervention;control group was added with 5μmol·L^(-1) dapagliflozin;25 mmol·L^(-1) glucose was added to model group;experimental group was added with 25 mmol·L^(-1) glucose and 5μmol·L^(-1) dapagliflozin,and cells in each group were cultured for 24 h for subsequent detection.Cell apoptosis was detected by flow cytometry,cell proliferation was detected by thiazolyl blue(MTT)method,and the levels of malondialdehyde(MDA)and reactive oxygen species(ROS)were detected by enzyme biochemistry and fluorescence microscopy.The activity of superoxide dismutase(SOD),TGF-β1(TGF-β1)and hepatocyte growth factor(HGF)protein levels in each group were determined by enzyme-linked immunosorbent assay.Results The apoptosis rates of HK-2 cells in blank group,control group,model group and experimental group were(4.73±0.46)%,(4.46±0.61)%,(35.42±2.37)%and(19.55±1.87)%;the optical density of cell proliferation were 0.52±0.09,0.49±0.07,0.13±0.03 and 0.29±0.05;MDA levels were(3.59±0.37),(4.13±0.52),(9.87±2.31)and(5.46±0.94)μmol·L^(-1);SOD levels were(3.75±0.31),(3.46±0.27),(2.03±0.16)and(3.12±0.22)kU·g HB-1;ROS levels were(3.12±0.55)%,(3.24±0.53)%,(14.67±2.64)%and(8.37±1.43)%;TGF-β1 levels were(446.31±55.42),(435.68±53.22),(1667.49±104.52)and(1235.74±95.35)pg·mg^(-1);HGF levels were(949.75±84.35),(964.84±68.26),(635.34±66.81)and(808.12±77.10)pg·mg^(-1).There were statistically significant differences in the above indicators between model group and blank group(all P<0.05),between experimental group and model group(all P<0.05).Conclusion Dapagliflozin can effectively inhibit the apoptosis of HK-2 cells induced by high glucose,promote hK-2 cells,and effectively reduce the levels of intracellular oxidative stress and markers of

关 键 词：达格列净 糖尿病肾病 细胞凋亡 细胞增殖 肾小管上皮细胞 

分 类 号：R97[医药卫生—药品]