不同药性成分对OAT4、URAT1抑制作用及对急性高尿酸血症小鼠血尿酸水平的影响
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摘要
目的研究不同药性中药成分对尿酸重吸收转运体尿酸转运蛋白1(URAT1)和有机阴离子转运蛋白4(OAT4)的抑制作用及对急性高尿酸血症小鼠尿酸水平的影响。方法应用OAT4、URAT1过表达单克隆细胞株(MDCK-hOAT4、HEK293-hURAT1),检测各药性中药单体对OAT4、URAT1的抑制作用及半数抑制浓度(IC50);运用黄嘌呤联合氧嗪酸钾制备急性高尿酸血症小鼠模型,研究原儿茶酸、甘草素、异甘草素对急性高尿酸血症小鼠血清尿酸的影响。结果苦味药中的川陈皮素,甘味药中的甘草素、异甘草素、甘草查耳酮A对OAT4抑制作用较强,IC50分别是0.556、18.40、6.831、6.825μmol/L。酸味药中的原儿茶酸、甘味药中的甘草素对URAT1具有较强的抑制作用,IC50分别是7.709、14.540μmol/L。甘草素能显著降低急性高尿酸血症小鼠的血尿酸水平,增加尿酸排泄量,降低血清肌酐、尿素氮的含量。结论川陈皮素、甘草素、异甘草素、甘草查耳酮A对OAT4具有较强的抑制作用,原儿茶酸、甘草素对URAT1具有较强抑制作用。甘草素能显著降低急性高尿酸血症小鼠血尿酸水平,其作用机制可能是抑制OAT4、URAT1对尿酸的重吸收,并具有一定的肾脏保护作用。
Objective To study the effects of Chinese herb ingredients with different properties on transporters(URAT1 and OAT4)involved in renal urate reabsorption and serum uric acid level in acute hyperuricemia mice. Methods The OAT4, URAT1-overexpressed monoclonal cell line(MDCK-hOAT4, HEK293-hURAT1) was constructed. The inhibition effect and the half maximal inhibitory concentration(IC50) of different ingredients to transport activity of OAT4 and URAT1 mediating 14 C-uric acid were determined. The effects of protocatechuic, liquiritigenin and isoliquiritigenin on serum uric acid levels in acute hyperuricemia mice were studied by the acute hyperuricemia mice induced by potassium oxonate and xanthine. Results The results indicated that nobiletin,liquiritigenin, isoliquiritigenin, licochalcone A with bitter flavor showed strong inhibition to OAT4. The IC50 of nobiletin,liquiritigenin, isoliquiritigenin, and licochalcone A on OAT4 were 0.556 μmol/L, 18.40 μmol/L, 6.831 μmol/L, and 6.825 μmol/L,respectively. Protocatechuic acid and liquiritigenin showed strong inhibition to URAT1 with IC50 of 7.709 μmol/L and 14.54 μmol/L,respectively. Liquiritigenin can significantly reduce the level of serum uric acid of acute hyperuricemia mice, increase the excretion of uric acid, and reduce the level of serum creatinine and blood urea nitrogen. Conclusion Nobiletin, liquiritigenin, isoliquiritigenin and licochalcone A can inhibit the transport activity of OAT4, while protocatechuic acid and liquiritigenin can inhibit the transport activity of URAT1. Liquiritigenin can significantly reduce the level of serum uric acid in acute hyperuricemia mice and protect kidney, the mechanism of which may be associated with the decreasing reabsorption of uric acid by inhibiting the activity of URAT1 and OAT4.
Objective To study the effects of Chinese herb ingredients with different properties on transporters(URAT1 and OAT4)involved in renal urate reabsorption and serum uric acid level in acute hyperuricemia mice. Methods The OAT4, URAT1-overexpressed monoclonal cell line(MDCK-hOAT4, HEK293-hURAT1) was constructed. The inhibition effect and the half maximal inhibitory concentration(IC50) of different ingredients to transport activity of OAT4 and URAT1 mediating 14 C-uric acid were determined. The effects of protocatechuic, liquiritigenin and isoliquiritigenin on serum uric acid levels in acute hyperuricemia mice were studied by the acute hyperuricemia mice induced by potassium oxonate and xanthine. Results The results indicated that nobiletin,liquiritigenin, isoliquiritigenin, licochalcone A with bitter flavor showed strong inhibition to OAT4. The IC50 of nobiletin,liquiritigenin, isoliquiritigenin, and licochalcone A on OAT4 were 0.556 μmol/L, 18.40 μmol/L, 6.831 μmol/L, and 6.825 μmol/L,respectively. Protocatechuic acid and liquiritigenin showed strong inhibition to URAT1 with IC50 of 7.709 μmol/L and 14.54 μmol/L,respectively. Liquiritigenin can significantly reduce the level of serum uric acid of acute hyperuricemia mice, increase the excretion of uric acid, and reduce the level of serum creatinine and blood urea nitrogen. Conclusion Nobiletin, liquiritigenin, isoliquiritigenin and licochalcone A can inhibit the transport activity of OAT4, while protocatechuic acid and liquiritigenin can inhibit the transport activity of URAT1. Liquiritigenin can significantly reduce the level of serum uric acid in acute hyperuricemia mice and protect kidney, the mechanism of which may be associated with the decreasing reabsorption of uric acid by inhibiting the activity of URAT1 and OAT4.
引文
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[13]牛艳芬,高丽辉,刘旭,等.芒果苷对氧嗪酸钾所致慢性高尿酸血症大鼠尿酸及肝肾功能的影响[J].中国药理学通报,2012,28(11):1578-1581.
[2]EleftheriadiS T,Golphinopoulos S,Pissas G,et al.Asymptomatic hyperuricemia and chronic kidney disease:Narrative review of a treatment controversial[J].J Adv Res,2017,8(5):555-560.
[3]刘金畅,王涛.治疗高尿酸血症药物研发现状与思考[J].天津中医药,2017,34(5):291-294.
[4]Burckhardt G.Drug transport by organic anion transporters(OATs)[J].Pharmacol Therap,2012,136(1):106-130.
[5]尧贵林,王海勇,陆涛.促尿酸排泄药物研究进展[J].中国药科大学学报,2016,47(4):491-496.
[6]李琳,张敏,李苒,等.中药治疗高尿酸血症实验研究进展[J].天津中医药,2013,30(3):190-192.
[7]韩彦琪,孟凡翠,许浚,等.基于网络药理学方法的元胡止痛滴丸治疗原发性痛经的配伍合理性研究[J].中草药,2017,48(3):526-532.
[8]李晓芳,张健康,王慧鸾,等.陈皮的研究进展[J].江西中医药,2014,45(3):76-78.
[9]姜雪,孙森凤,王悦,等.甘草药理作用研究进展[J].化工时刊,2017,31(7):25-28.
[10]Sun S,Wang K,Lei H,et al.Inhibition of organic cation transporter 2 and 3 may be involved in the mechanism of the antidepressant-like action of berberine[J].Prog Neuro Psychoph,2014,doi:10.1016/j.pnpbp.2013.11.005.
[11]Khanna D,Fitzgerald J D,Khanna P P,et al.2012American college of rheumatology guidelines for management of gout.part 1:Systematic nonpharmacologic and pharmacologic therapeutic approaches to hyperuricemia[J].Arthrit Care Res,2012,64(10):1431-1446.
[12]周盼,徐先祥,李沧海,等.中药干预尿酸转运体研究进展[J].中药药理与临床,2015,31(6):226-230.
[13]牛艳芬,高丽辉,刘旭,等.芒果苷对氧嗪酸钾所致慢性高尿酸血症大鼠尿酸及肝肾功能的影响[J].中国药理学通报,2012,28(11):1578-1581.