清热泄浊化瘀方对人肾小管上皮细胞UAT和URAT1基因表达的影响

Heat leakage for people removing stasis and turbidity tubular epithelial cells UAT and URAT1 gene expression

目的:观察不同浓度清热泄浊化瘀方(QRXZHYF)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:根据培养液中所含清热泄浊化瘀方浓度的不同,将HK-2细胞分为:(1)A组:DMEM/F-12+6mL/2kg清热泄浊化瘀方组;(2)B组:DMEM/F-12+12mL/2kg清热泄浊化瘀方组;(3)C组:DMEM/F-12+23.5mL/2kg清热泄浊化瘀方组;(4)对照组D组:仅使用DMEM/F-12组;每组均培养6瓶细胞。上述细胞在不同培养液中分别培养48h。采用实时荧光定量PCR检测HK-2细胞中hUAT mRNA,hURAT1 mRNA的相对表达量(2△△Ct法)。结果:所有标本均能检测到hUAT mRNA的表达,A、B、C组hUAT mRNA表达水平均明显高于对照组,(P<0.05),且其提高hUATmRNA水平程度呈剂量依赖性;各组间hURAT1 mRNA表达明显,A、B、C组与对照组比较,差异无统计学意义(P>0.05)。结论:不同浓度的清热泄浊化瘀方均可调节hUAT mRNA表达,且具有剂量依赖性;清热泄浊化瘀方对hURAT1 mRNA表达无调节作用。

Objective：To observe the different concentrations of heat Xiezhuo Huayu Recipe（QRXZHYF） whether the regulation of human renal tubular epithelial cells（HK-2 cells） urate transporter（hUAT） mRNA and organic anion transporter（hURAT1） mRNA expression.Methods：The culture medium contained in the heat of Xiezhuo Huayu different concentrations,the HK-2 cells were divided into：（1） A Group：DMEM/F-12＋6 mL/2 kg heat Xiezhuo treated group;（2） B group：DMEM/F-12＋12 mL/2 kg heat Xiezhuo treated group;（3） C groups：DMEM/F-12 ＋23.5 mL/2 kg heat Xiezhuo treated group;（4） control group D：Use only DMEM/F-12 group;each group of cells cultured for 6 bottles.The cells were cultured in different culture medium 48 h.Real-time fluorescence quantitative PCR detection of HK-2 cells hUAT mRNA,hURAT1 mRNA relative expression（2△△Ct method）.Results：All specimens hUAT mRNA could be detected in the expression,A,B,C group hUAT mRNA expression were significantly higher,（P〈0.05）,and the extent of its increase hUAT mRNA level dose-dependent manner;each group between hURAT1 mRNA expression was,A,B,C and control groups,the difference was not significant（P〉0.05）.Conclusion：Different concentrations of heat Xiezhuo Huayu Recipe can be adjusted hUAT mRNA expression in a dosedependent;heat Xiezhuo Huayu Recipe on hURAT1 mRNA expression was not regulation.