四妙散加味方对人肾小管上皮细胞UAT和URAT1基因表达的影响

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

目的:观察低、高剂量四妙散加味方(SMSJWF)是否调节人肾小管上皮细胞(HK-2细胞)尿酸盐转运子(hUAT)mRNA和有机阴离子转运体(hURAT1)mRNA的表达。方法:根据培养液中所含四妙散加味方剂量的不同,将HK-2细胞分为:DMEM/F-12+6 mL/2 kg四妙散加味方低剂量组;DMEM/F-12+12 mL/2 kg四妙散加味方高剂量组;DMEM/F-12+苯溴马隆4.8 mg/d阳性对照组;仅使用DMEM/F-12+空白血清对照组。四组细胞分别培养48 h,采用实时荧光定量PCR检测HK-2细胞中hUATmRNA、hURAT1 mRNA的相对表达量(2△△Ct法)。结果:所有标本均能检测到hUAT mRNA和hURAT1 mRNA的表达。低、高剂量组和阳性对照组hUAT mRNA表达水平均明显高于空白对照组(P<0.05);低剂量和阳性对照组的hURAT1 mRNA的表达明显低于空白对照组(P<0.05),而高剂量组hURAT1 mRNA的表达明显高于其他各组(P<0.05)。结论:低、高剂量四妙散加味方可上调hUAT mRNA表达;低剂量四妙散加味方与苯溴马隆均可下调hURAT1mRNA的表达,这可能是其降尿酸的作用机制之一。

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（P0.05）.Compared with blank control group,the expressions of hURAT1 mRNA in low dosage group and Benzbromarone group were reduced clearly（P0.05）.By contrast,the expression in high dosage group was increased clearly（P0.05）.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.