摘要
目的通过观察萆薢分清丸对高尿酸血症(HUA)模型大鼠肾脏尿酸盐转运蛋白及mRNA水平的影响,探讨该方对HUA大鼠的干预作用机制。方法60只雄性SD大鼠随机为正常组、模型组、别嘌醇组(0.03 g·kg^(-1))和萆薢分清丸低、中、高剂量组(0.8、1.6、3.2 g·kg^(-1))。除正常组外,其余各组大鼠每天灌胃氧嗪酸钾1.5 g·kg^(-1)和腺嘌呤0.1 g·kg^(-1)以建立HUA模型,连续28 d,以血尿酸(SUA)水平升高为标准。造模成功后依据组别给予相应药物干预,1次/d,连续28 d。末次给药24 h后,采集大鼠尿液和血液,采用酶比色法测定尿尿酸(UUA)、SUA、尿素氮(BUN)、血肌酐(SCr)水平;取大鼠双侧肾脏,称重,计算肾脏系数;苏木素-伊红(HE)染色法观察肾脏病理变化;免疫组化(IHC)检测肾脏组织中尿酸转运体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)、ATP结合盒转运蛋白G2(ABCG2)蛋白表达水平;实时荧光定量聚合酶链式反应(Real-time PCR)检测URAT1、GLUT9、OAT1、OAT3、ABCG2 mRNA水平。结果与正常组比较,模型组大鼠肾脏系数明显增加(P<0.05);SUA、BUN和SCr水平明显升高(P<0.05),UUA水平明显降低(P<0.05);肾小管代偿性扩张,管内可见尿酸盐结晶和蛋白管型;肾脏组织中URAT1、GLUT9蛋白表达水平及mRNA水平明显升高(P<0.05),OAT1、OAT3和ABCG2蛋白表达水平及mRNA水平明显降低(P<0.05)。与模型组比较,各给药干预组大鼠肾脏系数明显减小(P<0.05);SUA、BUN和SCr水平明显降低(P<0.05),UUA水平明显升高(P<0.05);肾脏组织病变明显减轻,URAT1、GLUT9蛋白表达水平及mRNA水平明显降低(P<0.05),OAT1、OAT3和ABCG2蛋白表达水平及mRNA水平明显升高(P<0.05)。结论萆薢分清丸可通过调节尿酸盐转运蛋白表达水平来实现其对HUA的干预作用。
Objective:To study the intervention effect of Bixie Fenqingwan on hyperuricemia(HUA rats by regulating urate transporters.Method:Sixty healthy rats were randomly divided into normal,model,allopurinol(0.03 g·kg^(-1)),and Bixie Fenqingwan low-,medium-and high-dose(0.8,1.6,3.2 g·kg^(-1))groups,10 in each group.Except the normal group,the other rats were given potassium oxonate 1.5 g·kg^(-1)and adenine0.1 g·kg^(-1)for 28 consecutive days to establish the HUA rat model,and rats with increased serum uric acid(SUA)were considered successfully modeled.After modeling,corresponding drugs were given to the groups,once per day.Urine and blood was collected after 24 h of the final administration.The levels of urine uric acid(UUA),SUA,blood urea nitrogen(BUN)and serum creatinine(SCr)were measured by enzymatic colorimetry.The rat kidneys were taken and weighed to calculate the kidney index.The pathological changes of kidney tissue were observed by haematoxylin-eosin(HE)staining.The protein and mRNA expressions of urate transporter 1(URAT1),glucose transporter 9(GLUT9),adenosine triphosphate-binding cassette transporter protein G2(ABCG2),organic anion transporter 1(OAT1)and organic anion 3 transporter(OAT3)in kidney tissue were detected by immunohistochemistry and real-time quantitative polymerase chain reaction(Real-time PCR),respectively.Result:Compared with the conditions in the normal group,the kidney index,levels of SUA,BUN and SCr,and protein and mRNA expressions of URAT1 and GLUT9 in kidney tissue were increased(P<0.05),while the UUA level and protein and mRNA expressions of OAT1,OAT3 and ABCG2 were decreased in the model group(P<0.05).In addition,there was compensatory dilatation with urate crystals and protein casts in renal tubules in the model group.Compared with the model group,the intervention groups had lowered kidney index(P<0.05),reduced levels of SUA,BUN and SCr(P<0.05),down-regulated protein and mRNA expressions of URAT1 and GLUT9(P<0.05),elevated UUA level(P<0.05)and up-regulated protein and mRNA expressions of OAT1,OAT3 and ABCG2(P<0.05),and the kidney tissue lesions were alleviated(P<0.05).Conclusion:Bixie Fenqingwan has intervention effect on HUA,and its mechanism may be related to regulating urate transporters.
作者
张明昊
杜婧雯
张童
郭申
俎兆轩
赵绅
王瑾瑾
ZHANG Minghao;DU Jingwen;ZHANG Tong;GUO Shen;ZU Zhaoxuan;ZHAO Shen;WANG Jinjin(School of Medicine,Henan University of Chinese Medicine,Zhengzhou 450046,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2023年第3期1-8,共8页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家自然科学基金项目(82104748)
河南省自然科学基金项目(202300410253)
河南省高等学校青年骨干教师培养计划项目(2020GGJS109)
河南中医药大学科研苗圃工程项目(MP2022-109)。
