高尿酸对血管内皮细胞增殖凋亡作用及其机制研究  

Effect of high urate on proliferation and apoptosis in human vascular endothelial cell and possible mechanism

作　　者：黄修献[1] 李海涛[1] HUANG Xiu-xian;LI Hai-tao(Department of Cardiovascular Medicine,Hainan General Hospital,Haikou 570000,Hainan Province,China)

机构地区：[1]海南省人民医院心血管内科

出　　处：《中国临床药理学杂志》2019年第22期2894-2897,共4页The Chinese Journal of Clinical Pharmacology

基　　金：海南省自然科学基金资助项目(2016CXTD012)

摘　　要：目的研究尿酸对人脐静脉内皮细胞株(ECV304)增殖凋亡的影响及可能的机制分析。方法(1)将人脐静脉内皮细胞株(ECV304)分为3组:溶剂对照组和低、高2个剂量实验组。2个剂量实验组加入10,20 mg·d L^-1的尿酸,溶剂对照组不作任何处理。(2)将ECV304分为4组:溶剂对照组、低剂量尿酸实验组(10 mg·d L^-1)、p38抑制剂(SB)组(SB,20μmol·L^-1)和联合组(尿酸10 mg·d L^-1+p38抑制剂20μmol·L^-1)。用CCK8检测给予尿酸后ECV304细胞增殖水平;用蛋白质印迹法检测半胱氨酸天冬氨酸蛋白酶-3(Caspase 3)、裂解的半胱氨酸天冬氨酸蛋白酶-3(Cleaved caspase-3)、p38丝裂原活化蛋白激酶(p38MAPK)和磷酸化-p38 MAPK(p-p38 MAPK)的蛋白水平。结果(1)在24 h后,溶剂对照组和低、高2个剂量实验组的细胞增殖率分别为(100.00±4.00)%,(80.52±3.11)%和(60.49±3.57)%;这3组的Caspase-3蛋白表达分别为0.99±0.11,1.26±0.15和1.59±0.13;这3组的Cleaved caspase-3的蛋白表达分别为0.98±0.14,1.70±0.10和2.19±0.11;这3组的p-p38 MAPK蛋白表达量为0.97±0.15,1.81±0.06和2.12±0.10;这3组的p38 MAPK蛋白表达量为1.30±0.11,1.00±0.10和0.88±0.04;上述指标:低、高2个剂量实验组与溶剂对照组相比,差异均有统计学意义(P<0.05或P<0.01)。(2)在24 h后,溶剂对照组、低剂量尿酸实验组、p38抑制剂组和联合组的细胞增殖率分别为(100.00±7.21)%,(79.61±3.34)%,(109.30±1.18)%和(90.69±1.89)%;这4组的Caspase-3的蛋白表达分别为1.24±0.04,1.63±0.06,1.04±0.04和1.29±0.03;这4组的Cleaved caspase-3的蛋白表达分别为0.93±0.05,1.49±0.12,0.73±0.05和0.97±0.05;这4组的p-p38蛋白的表达分别为1.16±0.07,1.56±0.07,0.89±0.06和1.02±0.05;这4组的p38蛋白的表达分别为1.27±0.12,0.98±0.02,0.88±0.03和0.69±0.05;3个处理组与溶剂对照组相比,Caspas-3表达差异均有统计学意义(均P<0.01);p38抑制剂组和联合组与溶剂对照组相比,Cleaved caspase-3和p38MAPK的表达差异均有�Objective To investigate the effect of high urate on human umbilical vein endothelial cell line(ECV304)and analyze its possible mechanism.Methods(1)ECV304 were divided into three groups:Solvent control group(without any treatment),experimental-L group and experimental-H groups(10,20 mg·L^-1 urate).(2)ECV304 were divided into 4 groups:Solvent control group(without any treatment),experimental-L group(10 mg·L^-1 urate),p38 inhibitor(SB)group(SB,20μmol·L^-1),united group(10 mg·L^-1 urate+20μmol·L^-1 SB).Cell Counting Kit 8 assay was used to test the proliferation level of urate on ECV304 cells.Western blot was employed to analyze the protein level of Caspase-3,Cleaved caspase-3,p38 mitogen-activated protein kinases(p38 MAPK)and phosphorylated-p38(p-p38).Results(1)After 24 h,the proliferation rate in solvent control group,experimental-L group and experimental-H group were(100.00±4.00)%,(80.52±3.11)%,(60.49±3.57)%,respectively;the expression of Caspase-3 in the three groups were 0.99±0.11,1.26±0.15,1.59±0.13,respectively;Cleaved caspase-3 in the three groups were 0.98±0.14,1.70±0.10,2.19±0.11 respectively;the expression of p-p38 MAPK in the three groups were 0.97±0.15,1.81±0.06,2.12±0.10;p38 MAPK protein level in the three groups were 1.30±0.11,1.00±0.10,0.88±0.04.Comparison between experimental-L and experimental-H groups with solvent control group,the difference of the factors were significant(all P<0.01).(2)After 24 h,the proliferation rate in solvent control group,experimental-L group,p38 inhibitor group,united group were(100.00±7.21)%,(79.61±3.34)%,(109.30±1.18)%,(90.69±1.89)%,respectively;the expression of Caspase-3 in the 4 groups were 1.24±0.04,1.63±0.06,1.04±0.04,1.29±0.03,respectively;the expression of Cleaved caspase-3 in the 4 groups were 0.93±0.05,1.49±0.12,0.73±0.05,0.97±0.05,respectively;the expression of p-p38 MAPK in the 4 groups were 1.16±0.07,1.56±0.07,0.89±0.06,1.02±0.05,respectively;the expression of p38 MAPK in the 4 groups were 1.27±0.12,0.98±0.02,0.88±0.0

关 键 词：尿酸 人脐静脉内皮细胞株 增殖 凋亡 p38丝裂原活化蛋白激酶信号通路 

分 类 号：R97[医药卫生—药品]