基于URAT1蛋白表达研究小檗碱对高尿酸血症小鼠炎症反应及氧化应激的影响

Effects of berberine on inflammatory reaction and oxidative stress in hyperuricemic mice based on URAT1 protein expression

目的 探索小檗碱对高尿酸血症小鼠炎症因子水平和氧化应激损伤的影响机制。方法 选取雄性SPF级昆明小鼠48只,分为对照组、模型组、苯溴马隆(20 mg/kg)组和小檗碱低、中、高剂量(50、100、200 mg/kg)组,每组8只。除对照组ig等量蒸馏水外,其余各组采用ig氧嗪酸钾和次黄嘌呤药物建立高尿酸血症小鼠模型,造模后苯溴马隆组ip给予20mg/kg苯溴马隆,小檗碱低、中、高剂量组分别ig 50、100、200 mg/kg小檗碱,各组均连续给药1周。检测小鼠血清指标尿酸、肌酐(Cr)水平;ELISA法测定C反应蛋白(CRP)水平、超氧化物歧化酶(SOD)的活性及丙二醛(MDA)的含量;苏木素–伊红染色观察小鼠肾脏病理组织变化情况;Western blotting法检测肾脏尿酸转运蛋白URAT1在小鼠体内的表达水平。结果 与对照组相比,模型组小鼠血清尿酸、Cr、CRP水平、MDA含量和URAT1蛋白表达均显著升高,SOD含量明显减少(P<0.01)。与模型组相比,苯溴马隆组和小檗碱50、100、200 mg/kg组小鼠血清尿酸、Cr、CRP水平、MDA含量和URAT1蛋白表达显著降低,SOD含量明显增加(P<0.05、0.01)。小鼠肾脏HE染色结果提示,与模型组相比,小檗碱50、100、200 mg/kg组肾脏病理性改变的程度均有所缓解。而小檗碱100、200 mg/kg组与苯溴马隆组比较无明显差异。结论小檗碱能降低高尿酸血症小鼠体内炎症水平,提高抗氧化能力。同时减轻高尿酸血症小鼠血清尿酸水平,其机制与抑制URAT1表达相关。

Objective To explore the mechanism and therapeutic effects of berberine on the levels of inflammatory factors and oxidative stress damage in mice with hyperuricemia. Methods Forty-eight male SPF grade Kunming mice were selected and divided into control group, model group, benbromarone(20 mg/kg) group, and berberine(50, 100, and 200 mg/kg) groups, with 8 mice in each group. Except for the control group, which was given the same amount of distilled water intragastrically, the other groups were ig administered with potassium oxazine and hypoxanthine to establish hyperuricemia mouse model. After modeling, benzbromarone group was ip 20 mg/kg benzbromarone, and berberine group was ig administered with 50, 100, 200 mg/kg berberine for 1 week,respectively. Serum uric acid and creatinine(Cr) levels were detected. The level of C-reactive protein(CRP), the activity of superoxide dismutase(SOD) and the content of malondialdehyde(MDA) were determined by ELISA. HE staining was used to observe the pathological changes of mouse kidney. The expression level of renal uric acid transporter URAT1 in mice was detected by Weste rn Blotting method. Results Compared with the control group, the levels of serum uric acid, Cr, CRP, MDA content, and URAT1 protein in model group were significantly increased, SOD content decreased obviously(P<0.01). Compared with model group, the levels of serum uric acid, Cr, CRP, MDA content, and URAT1 protein in benbromarone group and berberine 50, 100, and 200 mg/kg groups were significantly decreased, SOD content increased obviously(P<0.05, 0.01). The results of HE staining of mouse kidney indicated that compared with model group, berberine 50, 100, and 200 mg/kg groups had some relief in the degree of renal pathological changes.There was no significant difference between berberine 100 and 200 mg/kg groups and benzbromarone group. Conclusion Berberine can reduce inflammation and increase antioxidant capacity in hyperuricemia mice. At the same time, the serum uric acid level of hyperuricemia mice was reduced, and the mechanism was related to the inhibition of URAT1 expression.