玉米须黄酮类成分对HK-2细胞尿酸吸收及相关基因表达的影响

Effect of Total Flavonoids in Corn Stigma on Uric Acid Uptake and Related Gene Expression in HK-2 Cells

目的:探讨玉米须总黄酮对人肾小管上皮细胞凋亡及尿酸吸收的作用及其机制。方法:体外培养人肾小管上皮细胞(HK-2)进行尿酸吸收实验,考察各浓度玉米须总黄酮对HK-2细胞尿酸的吸收情况,同时,采用流式细胞术考察玉米须总黄酮对由尿酸诱导的HK-2细胞凋亡的影响;应用RT-PCR技术检测HK-2细胞中URAT1、GLUT9、OAT1、ABCG2相关基因表达情况。结果:与空白对照组相比,在给药孵育48 h后,玉米须总黄酮各浓度对HK-2细胞尿酸吸收均有不同程度的抑制作用,其中3 mg/mL浓度组抑制作用最强,其次为2.5 mg/ml,抑制率随着浓度的升高而升高。细胞凋亡实验显示,玉米须总黄酮1、1.5、3 mg/mL组能够降低尿酸诱导的HK-2细胞凋亡率(P<0.05或P<0.01);RT-PCR结果显示,与空白对照组比较,模型对照组PDZK1、URAT1、GLUT9的mRNA表达量明显上调(P<0.05),ABCG2、OAT1、OAT3 mRNA表达量明显下调(P<0.05或P<0.01);与模型对照组比较,玉米须总黄酮3 mg/mL、1.5 mg/mL、1.0 mg/mL组均能够显著下调PDZK1、URAT1、GLUT9mRNA的表达量(P<0.05或P<0.01);明显上调ABCG2、OAT1、OAT3 mRNA表达量(P<0.05或P<0.01)。结论:玉米须总黄酮能够抑制HK-2细胞对尿酸的吸收;抑制由尿酸诱导的HK-2细胞凋亡率,降低PDZK1、URAT1、GLUT9 mRNA基因表达水平;提高ABCG2、OAT1、OAT3 mRNA基因表达水平。

Objective:To investigate the effects of total flavonoids in stigma of corn(TFSC)on Human tubular epithelial cells(Human Kidney 2,HK-2)apoptosis and uric acid uptake.Methods:HK-2 cells were cultured in vitro for uric acid uptake experiment.The uric acid uptake in HK-2 cells was investigated under different doses of TFSC.Meanwhile,the apoptosis effect of TFSC on HK-2 cells was investigated by flow cytometry.RT-PCR was used to detect the genes expressions of URAT1,GLUT9,OAT1 and ABCG2 in HK-2 cells.Results:Compared with the control group,after incubation for 48h,all concentrations of TFSC inhibited the uptake of uric acid in HK-2 cells to different degrees,3mg/mL TFSC had the strongest inhibition effect,and 2.5 mg/ml TFSC had the second strongest inhibition effect.The inhibition rate increased with the increase of the concentration.The apoptosis experiments showed that,different doses of TFSC(1、1.5、3 mg/mL)reduced the apoptosis rate of uric acid induced HK-2 cells(P<0.05 or P<0.01).RT-PCR results showed that,compared with the control group,the expression levels of PDZK1,URAT1 and GLUT9 mRNA genes were significantly increased in the model group(P<0.05),the mRNA expressions of ABCG2、OAT1 and OAT3 were significantly decreased(P<0.05 or P<0.01).Compared with the model group,the mRNA expressions of PDZK1、URAT1 and GLUT9 were significantly decreased by TFSC(3 mg/mL、1.5 mg/mL、1.0 mg/mL)(P<0.05 or P<0.01),the mRNA expressions of ABCG2、OAT1 and OAT3 were significantly increased(P<0.05 or P<0.01).Conclusion:TFSC can inhibit the uptake of uric acid in HK-2 cells,it also inhibits the apoptosis rate of HK-2 cells induced by uric acid,reduces the expression levels of PDZK1,URAT1 and GLUT9 mRNA,and increase the mRNA expression levels of ABCG2,OAT1 and OAT3 genese.