目的 初步明確秋菊丸化學成分，研究秋菊丸對慢性高尿酸血癥（HUA）模型大鼠尿酸（UA）升高及腎臟損傷的治療作用及機制。方法 借助超高效液相色譜串聯(lián)四極桿靜電場軌道阱質(zhì)譜（UPLC-Q-Exactive Orbitrap MS）技術，初步鑒定秋菊丸的化學成分。48只雄性SD大鼠隨機分成對照組、模型組、苯溴馬隆（5.45 mg·kg^-1，陽性藥）組和秋菊丸低、中、高劑量（1.31、2.62、5.24 g·kg^-1）組，除對照組外，通過ig腺嘌呤和乙胺丁醇制備大鼠慢性HUA模型，造模同時每天ig給藥1次，連續(xù)28 d。使用全自動生化儀測定大鼠血清UA、肌酐（CRE）、尿素氮（BUN）水平；HE及Masson染色觀察腎臟組織病理學變化；實時熒光定量PCR（qRT-PCR）法檢測腎臟組織中葡萄糖轉(zhuǎn)運蛋白9 （GLUT9）、有機陰離子轉(zhuǎn)運體家族蛋白3 （OAT3）、ATP結(jié)合盒亞家族G成員2（ABCG2）、轉(zhuǎn)化生長因子-β（TGF-β1）、SMAD蛋白3（Smad3）、α-平滑肌肌動蛋白（α-SMA）的mRNA水平。結(jié)果 秋菊丸中共鑒定出41個成分，包括黃酮類（9個）、有機酸類（5個）、生物堿類（3個）、苯丙素類（7個）、香豆素類（1個）、酚類（3個）、呋喃類（2個）、脂肪酰類（6個）、醌類（2個）、萜類（1個）、內(nèi)酯類（1個）、脂肪酸類（1個）。給藥28 d后，HE染色結(jié)果表明，與模型組比較，各給藥組腎損傷的病理學改變明顯恢復，減輕了腎小球萎縮、腎小管擴張、腎纖維化及腎間質(zhì)炎性細胞浸潤等病理改變，其中秋菊丸高劑量組及苯溴馬隆組腎臟損傷最輕。與模型組比較，秋菊丸高劑量組的CRE、UA水平均顯著降低（P＜0.01），各劑量組BUN水平有降低趨勢，但無顯著性差異；高、中劑量組腎組織中腎小管上皮細胞中膠原蛋白的含量顯著降低（P＜0.05、0.01）；高劑量組GLUT9 mRNA表達量顯著減少（P＜0.05、0.01），各劑量組OAT3、ABCG2的mRNA表達量均顯著增加（P＜0.05、0.01）；中、高劑量α-SMA mRNA表達量顯著減少（P＜0.05、0.01），高劑量組TGF-β1 mRNA表達量顯著減少（P＜0.01），各給藥組Smad3的mRNA表達量有減少趨勢，但無顯著性差異。結(jié)論 初步明確秋菊丸化成成分，秋菊丸具有增強UA代謝、減輕HUA所致腎臟損傷及腎纖維化的作用，其機制可能與調(diào)控GLUT9-OAT3尿酸轉(zhuǎn)運體相關。

Objective 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 and α-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 epithelial cells in high and medium dose groups significantly decreased (P < 0.05, 0.01). Compared with model group, the expression level of GLUT9 mRNA was significantly reduced in the high-dose group (P < 0.05, 0.01), while the mRNA expression levels of OAT3 and ABCG2 were significantly increased in each dose group (P < 0.05, 0.01). Compared with model group, the α-SMA mRNA expression levels of medium and high-dose group were significantly reduced (P < 0.05, 0.01), while the TGF-β1 mRNA expression was significantly reduced (P < 0.01) in high-dose group. The mRNA expression levels of Smad3 in each treatment group showed a decreasing trend, but there was no significant difference. Conclusion Qiuju Wan can regulate uric acid transporters, thereby enhancing uric acid metabolism and alleviating kidney damage and renal fibrosis caused by high uric acid, and its mechanism may be related to regulation of GLUT9-OAT3 uric acid transporter.

R284.1；R285.5

國家中醫(yī)藥管理局古代經(jīng)典名方目錄定制（第二批民族醫(yī)藥專題GZY-KJS-2019-011）；天山英才項目（2022TSYCCX0021，2022TSYCLJ009）；中醫(yī)藥傳承與創(chuàng)新“百千萬”人才工程-青年岐黃學者