Effects of SiMiaoSanJiaWeiFang on the expression of human UAT and URAT1 transporter genes in HK-2 cells

Abstract

Abstract: AIM: To observe the effect of SiMiaoSanJiaWeiFang(SMSJWF) in different concentrations on the expression of human UAT and URAT1 transporter genes in HK-2 cells.METHODS: HK-2 cells were divided into 4 groups as follows: The low dosage group (cultured with DMEM/F-12 and 6 mL/2 kg SMSJWF ); the high dosage group (cultured with DMEM/F-12 and 12 mL/2 kg SMSJWF ); the positive control group(cultured with DMEM/F-4.8 mg/d Benzbromarone ); the blank control group(cultured with DMEM/F-12). In each group, 6 bottles of the cells were cultured in vitro for 48 hours. Then the relative quantity of hUAT and hURAT1 mRNA in the HK-2 cells was detected by real-time fluorescent quantitative PCR assay.RESULTS: hUAT and hURAT1 mRNA could be detected in all samples. And the level of hUAT mRNA in the blank control group were significantly lower than those in the other groups (P＜0.05). Compared with blank control group, the expressions of hURAT1 mRNA in low dosage group and Benzbromarone group were reduced clearly (P＜0.05). By contrast, the expression in high dosage group was increased clearly (P＜0.05).CONCLUSION: SiMiaoSanJiaWeiFang up-regulates the expression of hUAT gene. SiMiaoSanJiaWeiFang down-regulates the expression of hURAT1 gene, which may be one of the mechanism to reduce uric acid levels.

Key words: SiMiaoSanJiaWeiFang, Uric acid, Kidney tubules, Gene expression, Carrier proteins

CLC Number:

R311
R965.2

WANG Hua-jie, ZHANG Guo-hua, ZHOU-Zhen, FAN Xiu-zhen. Effects of SiMiaoSanJiaWeiFang on the expression of human UAT and URAT1 transporter genes in HK-2 cells[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2010, 15(9): 972-977.

References

[1] 王华杰, 张国华, 邹震, 等. 四妙散加味方治疗急性痛风性关节炎及高尿酸血症的临床研究[J].中国中医急症杂志, 2009, 18(10):1609-1610.
[2] Lipkowitz MS, Leal-Pinto E, Rappoport JZ, et al. Functional reconstitution, membrane targeting, genomic structure and chromosomal localization of a human urate transporter[J].J Clin Invest ,2001,107(9):1103-1115.
[3] Enomoto A, Niwa T, Kanai Y, et al. Urate transporter and renal hypouricemia[J]. Rinsho Byori, 2003, 51(9):892-897.
[4] Kawasugi K, Takeuchi F. PRPP synthetase superactivity[J]. NIPPON rinsho, 2003, 61(Suppl l):284-287.
[5] 李仪奎. 中药血清药理学实验方法的若干问题[J]. 中药新药与临床药学, 1999, 10(2):95- 98.
[6] Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^-ΔΔCt method[J]. Methods, 2001, 25(4):402-408.
[7] Lipkowitz MS, Leal-Pinto E, Cohen BE, et al. Galectin 9 is the sugar-regulated urate transporter/channel UAT[J]. Glycoconj, 2004, 19(7/8/9):491-498.
[8] Enomoto A, Kimura H, Chairoungdua A, et al. Molecular identification of a renal urate anion exchanger that regulates blood urate levels[J]. Nature, 2002, 417(6887):447-452.
[9] Ichida K, Hosoyamada M, Hisatome I, et al. Clinical and molecular analysis of patients with renal hypouricemia in Japan-influence of URAT1 gene on urinary urate excretion[J]. J Am Soc Nephrol, 2004, 15(1):164-173.
[10] 王浴生, 邓文龙, 薛春生. 中药药理与应用[M]. 2版.北京:人民卫生出版社, 1998: 64-65.
[11] 王本祥. 现代中药药理学[M].天津: 天津科学技术出版社, 1997:403.
[12] 王效玲, 刘平, 刘成海, 等. 丹酚酸A对成纤维细胞活力, 增殖及胶原合成的影响[J]. 中西医结合肝病杂志, 2000, 10(1):24-25.
[13] 王海东, 葛飞, 郭玉松, 等. 金钱草提取物对高尿酸血症小鼠的影响 [J]. 中国中药杂志, 2002, 27(12):939-941.
[14] 孙晓波, 代竹青, 李毅, 等. 藏药桑当抗痛风的实验研究[J]. 四川中医, 2005, 23(6): 19-20.
[15] 吕耀中, 胡庆华, 王星, 等. 二妙丸水提取物对高尿酸血症小鼠尿酸失衡及其相关基因和蛋白水平的影响[J]. 中草药, 2010, 41(3):418-423.
[16] Bleasby K, Hall LA, Perry JL, et al. Functional consequences of single nucleotide polymorphisms in the human organic anion transporter hOAT1(SLC22A6)[J]. J Pharmacol Exp Ther,2005, 314(2):923-931.
[17] Erdman AR, Mangravte LM, Urban TJ, et al. The human organic anion transporter 3(OAT3;SLC22A8):genetic variation and functional genomics[J]. Am J Physiol Renal Physiol, 2006, 290(4):F905-F912.