秋菊丸化学成分鉴定及靶向GLUT9-OAT3尿酸转运体介导慢性高尿酸血症大鼠尿酸代谢和肾保护的机制研究  

Identification of chemical constituents of Qiuju Wan and study on mechanism of targeting GLUT9-OAT3 uric acid transporter mediating uric acid metabolism and renal protection in chronic hyperuricemia rats

作　　者：努尔玛娜提·胡安别克 周子虔 王春 麦丽克姆·麦图如孜 哈木拉提·哈斯木 都田芳 祖力皮卡尔·吾斯曼 蔡晓霞 霍仕霞 李治建 斯拉甫·艾白 HUANBIEKE Nuermanati;ZHOU Ziqian;WANG Chun;MATTURUZI Malikam;HASIMU Hamulati;DU Tianfang;WUSIMAN Zulipikaer;CAI Xiaoxia;HUO Shixia;LI Zhijian;AIBAI Silafu(College of Pharmacy,Xinjiang Medical University,Urumqi 830011,China;Xinjiang Uygur Autonomous Region Uygur Medical Research Institute,Urumqi 830011,China;Department of Pharmacology,Xinjiang Institute of Materia Medica,Urumqi 830011,China;Xinjiang Uygur Autonomous Region Uygur Hospital,Urumqi 830049,China;Hubei Topgene Biotechnology Co.,Ltd.,Wuhan Branch,Wuhan 430075,China)

机构地区：[1]新疆医科大学药学院,新疆乌鲁木齐830011 [2]新疆维吾尔自治区维吾尔医药研究所,新疆乌鲁木齐830011 [3]新疆维吾尔自治区药物研究所药理研究室,新疆乌鲁木齐830011 [4]新疆维吾尔自治区维吾尔医医院,新疆乌鲁木齐830049 [5]湖北天勤生物科技有限公司武汉分公司,湖北武汉430075

出　　处：《药物评价研究》2023年第6期1201-1213,共13页Drug Evaluation Research

基　　金：国家中医药管理局古代经典名方目录定制(第二批民族医药专题GZY-KJS-2019-011);天山英才项目(2022TSYCCX0021,2022TSYCLJ009);中医药传承与创新“百千万”人才工程-青年岐黄学者。

摘　　要：目的初步明确秋菊丸化学成分,研究秋菊丸对慢性高尿酸血症(HUA)模型大鼠尿酸(UA)升高及肾脏损伤的治疗作用及机制。方法借助超高效液相色谱串联四极杆静电场轨道阱质谱(UPLC-Q-Exactive Orbitrap MS)技术,初步鉴定秋菊丸的化学成分。48只雄性SD大鼠随机分成对照组、模型组、苯溴马隆(5.45 mg·kg^(-1),阳性药)组和秋菊丸低、中、高剂量(1.31、2.62、5.24 g·kg^(-1))组,除对照组外,通过ig腺嘌呤和乙胺丁醇制备大鼠慢性HUA模型,造模同时每天ig给药1次,连续28 d。使用全自动生化仪测定大鼠血清UA、肌酐(CRE)、尿素氮(BUN)水平;HE及Masson染色观察肾脏组织病理学变化;实时荧光定量PCR(qRT-PCR)法检测肾脏组织中葡萄糖转运蛋白9(GLUT9)、有机阴离子转运体家族蛋白3(OAT3)、ATP结合盒亚家族G成员2(ABCG2)、转化生长因子-β(TGF-β1)、SMAD蛋白3(Smad3)、α-平滑肌肌动蛋白(α-SMA)的mRNA水平。结果秋菊丸中共鉴定出41个成分,包括黄酮类(9个)、有机酸类(5个)、生物碱类(3个)、苯丙素类(7个)、香豆素类(1个)、酚类(3个)、呋喃类(2个)、脂肪酰类(6个)、醌类(2个)、萜类(1个)、内酯类(1个)、脂肪酸类(1个)。给药28 d后,HE染色结果表明,与模型组比较,各给药组肾损伤的病理学改变明显恢复,减轻了肾小球萎缩、肾小管扩张、肾纤维化及肾间质炎性细胞浸润等病理改变,其中秋菊丸高剂量组及苯溴马隆组肾脏损伤最轻。与模型组比较,秋菊丸高剂量组的CRE、UA水平均显著降低(P<0.01),各剂量组BUN水平有降低趋势,但无显著性差异;高、中剂量组肾组织中肾小管上皮细胞中胶原蛋白的含量显著降低(P<0.05、0.01);高剂量组GLUT9 mRNA表达量显著减少(P<0.05、0.01),各剂量组OAT3、ABCG2的mRNA表达量均显著增加(P<0.05、0.01);中、高剂量α-SMA mRNA表达量显著减少(P<0.05、0.01),高剂量组TGF-β1 mRNA表达量显著减少(P<0.01),各给药组Smad3Objective To preliminarily clarify the chemical composition of Qiuju Wan,and to study the therapeutic effect and mechanism of Qiuju Wan on the elevation of uric acid(UA)and kidney damage in chronic hyperuricemia(HUA)model rats.Methods Ultra-performance liquid chromatography tandem quadrupole electrostatic field,orbital well mass spectrometry(UPLC-QExactive Orbitrap MS)technology was used to preliminarily identify the chemical composition of Qiuju Wan.Forty-eight SD rats(males)were randomly divided into controlgroup,model group,benzbromarone(5.45 mg·kg^(-1)),and QJW low,medium,high dose(1.31,2.62,and 5.24 g·kg^(-1))group.Except for control group,the rat chronic HUA model was prepared by adenine and ethambutol intragastrically,which was simultaneously given intragastrically once a day for consecutive 28 days.Biochemical parameters such as UA,creatinine(CRE)and urea nitrogen(BUN)were determined,and histopathological changes of kidney were observed by HE and Masson staining.Gene levels of GLUT9,OAT3,ABCG2,TGF-β1,Smad3 andα-SMA in kidney tissue were detected by qRT-PCR.Results A total of 41 components were identified in Qiuju Wan,including flavonoids(9),organic acids(5),alkaloids(3),phenylpropanoids(7),coumarins(1),phenols(3),furans(2),fatty acyls(6),quinones(2),terpenoids(1),lactones(1),and fatty acids(1).After 28 days of administration,HE staining results showed that compared with the model group,the pathological changes of renal damage in each treatment group significantly recovered,reducing pathological changes such as glomerular atrophy,renal tubular dilation,renal fibrosis,and interstitial inflammatory cell infiltration.Among them,the high-dose group of Qiuju Wan and the group of benzbromarone had the least renal damage.Compared with model group,the CRE and UA levels in the high-dose group of Qiuju Wan were significantly reduced(P<0.01),and the BUN levels in each dose group showed a decreasing trend,but there was no significant difference.Compared with model group,the content of collagen in renal tubular epi

关 键 词：秋菊丸 成分 黄酮类 有机酸类 生物碱类 苯丙素类 尿酸 高尿酸血症 肾损伤 转运体 

分 类 号：R284.1[医药卫生—中药学] R285.5[医药卫生—中医学]