木蝴蝶醇提物对高尿酸血症小鼠的降尿酸及肾保护作用

Effect of Alcohol Extract of Oroxylum indicum on Reducing Uric Acid and Protecting Kidney in Hyperuricemia Mice

目的:建立高尿酸血症(HUA)体内模型,考察木蝴蝶醇提物(MHD-80)降尿酸及肾脏保护作用。方法:采用氧嗪酸钾(350 mg·kg^(-1))及腺嘌呤(80 mg·kg^(-1))构建小鼠HUA体内模型评价MHD-80的降尿酸相关机制及肾功能保护作用,将70只雄性ICR小鼠随机分为正常组、模型组、别嘌呤醇组(5 mg·kg^(-1))、非布索坦组(5 mg·kg^(-1))、MHD-80低、中、高剂量组(3、6、12 mg·kg^(-1))7组,每组10只。除正常组,其余组均连续14 d灌胃氧嗪酸钾及腺嘌呤构建HUA模型。造模第8~14天,每组灌胃给予相应药物,每天1次,末次给药后1 h,摘眼球取血,并收集小鼠肾脏及肝脏组织。酶比色法检测血清中尿酸(UA)、尿素氮(BUN)、肌酐(Cr)水平及肝脏中黄嘌呤氧化酶(XO)活性,酶联免疫吸附测定法(ELISA)检测小鼠血清中肿瘤坏死因子-α(TNF-α)及白细胞介素-1β(IL-1β)含量,苏木素-伊红(HE)染色观察肾脏组织病理改变,蛋白免疫印迹法(Western blot)检测肾脏组织ATP结合盒转运蛋白G2(ABCG2)及葡萄糖易化转运蛋白9(GLUT9)蛋白表达水平。结果:体内实验结果显示,与正常组比较,模型组小鼠血清中UA、Cr、BUN水平、炎症因子TNF-α、IL-1β及肝脏XOD活性明显升高(P<0.05,P<0.01),肾脏组织中GLUT9蛋白表达明显上调(P<0.05),ABCG2蛋白表达明显下调(P<0.05),肾脏出现明显损伤;与模型组比较,MHD-80中、高剂量组小鼠血清中UA、BUN、Cr、TNF-α、IL-1β水平及肝脏XOD活性有不同程度降低(P<0.05,P<0.01),MHD-80高剂量组的GLUT9蛋白表达显著下调(P<0.01)、ABCG2蛋白表达明显上调(P<0.05),MHD-80组肾损伤程度减轻。结论:MHD-80具有一定的降尿酸、抗炎及抗肾损伤作用,其作用与抑制XOD活性、调节ABCG2及GLUT9尿酸转运蛋白表达有关。

Objective:To investigate the effect of ethanol extract of Oroxylum indicum(MHD-80)on uric acid reduction and renal protection in hyperuricemia(HUA)model.Method:Potassium oxazine(350 mg·kg^(-1))and adenine(80 mg·kg^(-1))were used to construct a mouse model of HUA in vivo to evaluate the mechanism related to uric acid lowering and the protective effect of renal function of MHD80.Seventy male ICR mice were randomly divided into 7 groups(n=10),including normal group,model group,allopurinol group(5 mg·kg^(-1)),febusotan group(5 mg·kg^(-1)),low(3 mg·kg^(-1)),medium(6 mg·kg^(-1))and high(12 mg·kg^(-1))dose groups.Except for normal group,the other groups were intragastric administration of potassium oxyazinate and adenine for 14 consecutive days to establish HUA model.On the 8th to 14th day after modeling,each group was given corresponding drugs by intragastric administration,once a day.1 h after the last administration,blood was collected from eyeballs,and kidney and liver tissues of mice were collected.Serum levels of uric acid(UA),urea nitrogen(Cr)and creatinine(BUN)and liver activity of xanthine oxidase(XOD)were determined by enzyme colorimetry.Serum contents of tumor necrosis factorα(TNF-α)and interleukin1β(IL-1β)were determined by enzyme-linked immunosorbent assay(ELISA).Hematoxilin-eosin(HE)staining was used to observe the pathological changes of kidney tissue.The expression levels of ATP-binding box transporter G2(ABCG2)and glucose-facilitating transporter-9(GLUT9)in kidney tissue were detected by Western blot.Result:In vivo experiment,compared with normal group,the serum levels of UA,Cr,BUN,inflammatory factors TNF-α,IL-1βand liver XOD activity in model group were significantly increased(P<0.05,P<0.01),and the expression of GLUT9 protein in kidney tissue was significantly up-regulated(P<0.05).ABCG2 protein expression was significantly down-regulated(P<0.05),and kidney injury was obvious.Compared with model group,the levels of UA,BUN,Cr,TNF-α,IL-1βand the activity of liver XOD in serum of mice in mediumdose and high-dose groups of MHD-80 were decreased to different degrees(P<0.05,P<0.01),GLUT9 protein expression was significantly down-regulated and ABCG2 protein expression was significantly up-regulated(P<0.05,P<0.01)in high-dose group of MHD-80,and the degree of kidney injury was reduced in group of MHD-80.Conclusion:MHD-80 has certain uric acid lowering,anti-inflammatory and anti-renal injury effects,which are related to inhibiting XOD activity and regulating the expression of ABCG2 and GLUT9 uric acid transporter.