水蛭素对大鼠NRK-52E细胞尿酸盐转运体表达的影响  

Effect of Hirudin on the Expression of Urate Transporter in NRK-52E Cells

作　　者：赵应学[1] 黄思诗 梁红[2] 吴林秀[2] 梁杏梅[2] 周维海 甄汉深 黄敏琪[2] 刘喜华[2] Zhao Yingxue;Huang Sishi;Liang Hong;Wu Linxiu;Liang Xingmei;Zhou Weihai;Zhen Hanshen;Huang Minqi;Liu Xihua(Guangxi Jiangbin Hospital,Nanning 530021,China;Guangxi Medical College,Nanning 530023,China;Guangxi Kekang Technology Group Co.,Ltd.,Nanning 530001,China;College of Medicine,Guangxi University of Chinese Medicine,Nanning 530001,China)

机构地区：[1]广西壮族自治区江滨医院,南宁530021 [2]广西卫生职业技术学院,南宁530023 [3]广西科康科技集团有限公司,南宁530001 [4]广西中医药大学药学院,南宁530001

出　　处：《世界科学技术-中医药现代化》2021年第6期1972-1977,共6页Modernization of Traditional Chinese Medicine and Materia Medica-World Science and Technology

基　　金：广西壮族自治区自然科学基金委员会面上项目(2018GXNSFAA281093):基于尿酸转运子的水蛭素抗高尿酸血症分子机制研究;负责人:刘喜华。

摘　　要：目的探讨金边蚂蟥活性成分水蛭素对大鼠NRK-52E细胞尿酸盐转运体表达的影响。方法采用不同浓度(0、0.01、0.1、1、5和10mg·mL^(-1))水蛭素处理大鼠NRK-52E细胞,细胞计数试剂盒-8(Cell counting kit-8,CCK8)检测水蛭素对细胞的毒性作用。将细胞分为正常组、黄嘌呤组(200μmol·L^(-1))、水蛭素低(0.01 mg·mL^(-1))、中(0.1 mg·mL^(-1))、高浓度组(5 mg·mL^(-1)),培养24 h后,Hoechst33258染色观察细胞形态,实时荧光定量PC R(Real-time quantitative polymerase chain reaction,RT-qPC R)和蛋白免疫印迹法(Western blot)检测细胞中葡萄糖转运蛋白9(Glucose transporter 9,GLUT9)、有机阴离子运输蛋白(Organic anion transporter 1,OAT1)和尿酸盐转运体1(Urate transporter 1,URAT1)mRNA及蛋白表达水平。结果水蛭素对大鼠NRK-52E细胞毒性作用较小。与正常组比较,黄嘌呤组细胞碎片增多,细胞中GLUT9和URAT1mRNA及蛋白表达均显著升高(P<0.05),而OAT1 mRNA和蛋白表达均显著降低(P<0.05)。与黄嘌呤组比较,水蛭素组细胞碎片减少,GLUT9和URAT1表达显著降低(P<0.05),OAT1表达显著升高(P<0.05),且高浓度组比低浓度组更明显(P<0.05)。结论水蛭素可能通过调节肾细胞尿酸盐转运体(GLUT9、URAT1、OAT1)的表达对尿酸排泄的发挥作用。Objective To investigate the effect of hirudin on the expression of urate transporter in NRK-52 E cells.Methods NRK-52 E cells were treated with hirudin at different concentrations(0,0.01,0.1,1,5 and 10 mg·mL^(-1)),and the toxicity of hirudin was evaluated by the cell counting kit-8 assay.NRK-52 E cells were divided into five groups:normal,xanthine(200μmol·L^(-1)),and hirudin low(0.01 mg·mL^(-1)),medium(0.1 mg·mL^(-1)),and high(5 mg·mL^(-1))concentration.After 24 hours of culture,cell morphology was observed by Hoechst33258 staining,and the mRNA and protein expression levels of glucose transporter 9(GLUT9),organic anion transporter 1(OAT1),and urate transporter 1(URAT1)were detected by real-time quantitative polymerase chain reaction(RT-qPCR)and western blot,respectively Results Hirudin exerted negligible toxic effect on NRK-52 E cells.Compared with the normal group,xanthine group showed an increase in the number of cell fragments,significantly increased mRNA and protein levels of GLUT9 and URAT1(P<0.05),and significantly decreased mRNA and protein levels of OAT1(P<0.05).Compared with the xanthine group,the hirudin group showed significantly decreased expression of GLUT9 and URAT1(P<0.05),while that of OAT1 increased significantly(P<0.05).The effect of hirudin was more prominent at high concentration than at low concentration(P<0.05).Conclusion Hirudin may play a role in uric acid excretion by regulating the expression of urate transporters(GLUT9,URAT1,and OAT1)in renal cells.

关 键 词：水蛭素 NRK-52E细胞 尿酸盐转运体 

分 类 号：R285.5[医药卫生—中药学]