背景:高尿酸血症是痛风的重要病理基础,也是代谢综合征和心血管和慢性肾脏疾病的独特危险因素,但目前缺乏安全有效的治疗方法。Paeonia×suffruticosaAndrewsleaf有效降低痛风患者的血清尿酸;然而,物质基础和机制尚不清楚。
目的:研究山竹叶在高尿酸血症小鼠体内的主要活性成分及其作用机制。
方法:使用高效液相色谱分析鉴定了P.suffruticosa叶的化学成分。在高嘌呤饮食14天诱导的高尿酸血症小鼠中评估了苦参叶提取物(12.5、25、50、100和200mg/kg)及其成分的抗高尿酸血症活性。然后,在另一个高尿酸血症小鼠模型中评估了芹菜素7-O-葡萄糖苷(0.09,0.18和0.36mg/kg)的尿酸盐降低作用,该模型通过给予氧酸盐钾和腺嘌呤4周建立.通过研究黄嘌呤氧化酶在体外,血清和肝脏中的活性以及通过分子对接,阐明了芹菜素7-O-葡萄糖苷对尿酸产生的抑制作用。免疫荧光和蛋白质印迹分析肾尿酸转运蛋白1(URAT1)的表达,葡萄糖转运蛋白9(GLUT9),有机阴离子转运蛋白1(OAT1),和ATP结合盒G成员2(ABCG2)蛋白阐明了芹菜素7-O-葡萄糖苷如何促进尿酸排泄。
结果:在P.suffruticosaleaf中鉴定出六种化合物:没食子酸,没食子酸甲酯,氧合烟苷,芍药苷,胆酰芍药苷,和芹菜素7-O-葡萄糖苷。P.suffruticosa叶提取物显着减弱增加的血清尿酸,肌酐,和高尿酸血症小鼠的黄嘌呤氧化酶活性。来自P.suffuticosa叶的芹菜素7-O-葡萄糖苷降低尿酸,肌酐,和血清丙二醛水平,增加超氧化物歧化酶活性,部分恢复了高尿酸血症小鼠的脾脏系数。芹菜素7-O-葡萄糖苷在体外抑制黄嘌呤氧化酶活性,并降低高尿酸血症小鼠的血清和肝脏黄嘌呤氧化酶活性和肝脏黄嘌呤氧化酶蛋白表达。分子对接显示芹菜素7-O-葡萄糖苷与黄嘌呤氧化酶结合。芹菜素7-O-葡萄糖苷通过调节肾尿酸转运蛋白URAT1、GLUT9、OAT1和ABCG2促进尿酸排泄。芹菜素7-O-葡萄糖苷通过降低血清肌酐来保护高尿酸血症引起的肾损伤和氧化应激,血尿素氮,丙二醛,和肾脏活性氧水平;增加血清和肾脏超氧化物歧化酶活性;恢复肾脏系数;减少肾脏病理损伤。
结论:芹菜素7-O-葡萄糖苷是高尿酸血症小鼠的主要降尿酸活性成分。它抑制肝脏黄嘌呤氧化酶活性以减少尿酸合成,并调节肾尿酸转运蛋白以刺激尿酸排泄,减轻高尿酸血症引起的肾脏损害。
BACKGROUND: Hyperuricemia is an important pathological basis of gout and a distinct hazard factor for metabolic syndromes and cardiovascular and chronic renal disease, but lacks safe and effective treatments currently. Paeonia × suffruticosa Andrews leaf effectively reduced serum uric acid in gout patients; however, the material foundation and the mechanism remain unclear.
OBJECTIVE: To determine the primary active components and mechanism of P. suffruticosa leaf in hyperuricemic mice.
METHODS: The chemical constituents of P. suffruticosa leaf was identified using high-performance liquid chromatographic analysis. The anti-hyperuricemic activity of P. suffruticosa leaf extract (12.5, 25, 50, 100, and 200 mg/kg) and its components was evaluated in hyperuricemic mice induced by a high purine diet for 14 days. Then, the urate-lowering effects of apigenin 7-O-glucoside (0.09, 0.18, and 0.36 mg/kg) were assessed in another hyperuricemic mice model built by administrating potassium oxonate and adenine for 4 weeks. The inhibitory effect of apigenin 7-O-glucoside on uric acid production was elucidated by investigating xanthine oxidase activity in vitro and in serum and the liver and through molecular docking. Immunofluorescence and western blot analyses of the expression of renal urate transporter 1 (URAT1), glucose transporter 9 (GLUT9), organic anion transporters 1 (OAT1), and ATP-binding cassette G member 2 (ABCG2) proteins elucidated how apigenin 7-O-glucoside promoted uric acid excretion.
RESULTS: Six compounds were identified in P. suffruticosa leaf: gallic acid, methyl gallate, oxypaeoniflorin, paeoniflorin, galloylpaeoniflorin, and apigenin 7-O-glucoside. P. suffruticosa leaf extract significantly attenuated increased serum uric acid, creatinine, and xanthine oxidase activity in hyperuricemic mice. Apigenin 7-O-glucoside from P. suffruticosa leaf reduced uric acid, creatinine, and malondialdehyde serum levels, increased superoxide dismutase activity, and partially restored the spleen coefficient in hyperuricemic mice. Apigenin 7-O-glucoside inhibited xanthine oxidase activity in vitro and decreased serum and liver xanthine oxidase activity and liver xanthine oxidase protein expression in hyperuricemic mice. Molecular docking revealed that apigenin 7-O-glucoside bound to xanthine oxidase. Apigenin 7-O-glucoside facilitated uric acid excretion by modulating the renal urate transporters URAT1, GLUT9, OAT1, and ABCG2. Apigenin 7-O-glucoside protected against renal damage and oxidative stress caused by hyperuricemia by reducing serum creatinine, blood urea nitrogen, malondialdehyde, and renal reactive oxygen species levels; increasing serum and renal superoxide dismutase activity; restoring the renal coefficient; and reducing renal pathological injury.
CONCLUSIONS: Apigenin 7-O-glucoside is the main urate-lowering active component of P. suffruticosa leaf extract in the hyperuricemic mice. It suppressed liver xanthine oxidase activity to decrease uric acid synthesis and modulated renal urate transporters to stimulate uric acid excretion, alleviating kidney damage caused by hyperuricemia.