摘要
目的:从肾脏尿酸盐转运体角度探讨痛风宁降低血尿酸的机制。方法:将人肾小管上皮细胞(HK-2)随机分为正常组,模型组,痛风宁低、中、高剂量组(7. 65,15. 3,30. 6 g·kg-1),苯溴马隆组(50μmo1·L-1),分别予不同培养液进行干预。于干预24 h后收集HK-2及上清液,采用蛋白免疫印迹法(Western blot),实时荧光定量聚合酶链式反应(Real-time PCR)检测各组HK-2中尿酸盐转运蛋白1(URAT1),葡萄糖转运体9(GLUT9),有机阴离子转运体1(OAT1),有机阴离子转运体3(OAT3)和三磷酸腺苷结合盒转运蛋白G2(ABCG2)蛋白及mRNA的表达。结果:与正常组比较,模型组URAT1,GLUT9蛋白及mRNA表达显著升高(P <0. 01),ABCG2蛋白及mRNA表达显著下降(P <0. 01);与模型组比较,痛风宁各剂量组和苯溴马隆组URAT1,GLUT9蛋白及mRNA表达均明显下降(P <0. 01),且痛风宁各剂量组优于苯溴马隆组,痛风宁各剂量组ABCG2蛋白及mRNA表达显著升高(P <0. 01);与痛风宁中剂量组比较,痛风宁低、高剂量组URAT1,GLUT9蛋白及mRNA表达明显升高,ABCG2蛋白及mRNA表达明显下降(P <0. 01)。OAT1和OAT3蛋白及mRNA于各组均无表达。结论:痛风宁可通过下调HK-2中URAT1,GLUT9蛋白及mRNA表达,上调ABCG2蛋白及mRNA表达,调节肾小管对尿酸的重吸收和分泌,促进尿酸在肾脏的排泄,降低血尿酸水平。提示对肾脏尿酸转运体蛋白的调节可能是痛风宁发挥利湿排浊功效而降低血尿酸的具体机制之一,OAT1和OAT3蛋白及mRNA在体外培养的HK-2中无表达。
Objective:Study on the mechanism of Tongfengning in reducing serum uric acid from the perspective of renal urate transporter.Method:The human renal tubular epithelial cells(HK-2)was randomly divided into normal group,model group,Tongfengning low,medium and high dose group(7.65,15.3,30.6 g·kg-1)and benzbromarone group(50μmo1·L-1),different culture media were given for intervention.HK-2 and cell supernatant were collected after 24 h of intervention.The expressions of urate transporter 1(URAT1),glucose transporter 9(GLUT9),organic anion transporter 1(OAT1),organic anion transporter 3(OAT3),and ATP-binding cassette superfamily G member 2(ABCG2)protein and mRNA were detected in HK-2 of all groups by Western blot and Real-time PCR.Result:Compared with normal group,the expression of URAT1,GLUT9 protein and mRNA was significantly increased(P<0.01),while the expression of ABCG2 protein and mRNA was significantly decreased in model group(P<0.01).Compared with model group,the expression of URAT1,GLUT9 protein and mRNA in each dose of Tongfengning group and benzbromarone group were decreased(P<0.01),and each doses of Tongfengning group was superior to the benzbromarone group.The expression of ABCG2 protein and mRNA was increased in each dose of Tongfengning group(P<0.01).Compared with Tongfengning medium dose group,the expression of URAT1,GLUT9 protein and mRNA increased,while the expression of ABCG2 protein and mRNA decreased in the low and Tongfengning high dose groups(P<0.01).OAT1 and OAT3 were not expressed in all groups.Conclusion:Tongfengning can regulate the reabsorption and secretion of uric acid in renal tubules,promote the excretion of uric acid in kidney and reduce the level of serum uric acid by down-regulating the expression of URAT1,GLUT9 protein and mRNA in HK-2 and up-regulating the expression of ABCG2 protein and mRNA.It is suggested that the regulation of renal uric acid transporter protein may be one of the specific mechanisms of Tongfengning to reduce serum uric acid by promoting dampness and turbid re
作者
李保林
王建辉
蔡唐彦
郭洁梅
滕方舟
朱亚菊
林建平
毛骁
肖艳
苏友新
LI Bao-lin;WANG Jian-hui;CAI Tang-yan;GUO Jie-mei;TENG Fang-zhou;ZHU Ya-ju;LIN Jian-ping;MAO Xiao;XIAO Yan;SU You-xin(College of Traditional Chinese Medicine(TCM),College of Rehabilitation Medicine,Fujian University of TCM,Fuzhou 350122,China;Fuijian Health College,Fuzhou 350101,China;College of Medical Technology and Engineering,Fujian Medical University,Fuzhou 350004,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2019年第21期53-59,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(81473495)
