摘要目的 观察痛风患者与健康人群血清中雌二醇水平差异,雌二醇对人肾小管上皮细胞(HK-2)尿酸转运体ABC转运蛋白2(ABCG2)表达的影响,并研究磷脂酰肌醇-3-激酶/蛋白激酶B(PI3K/Akt)通路在雌二醇调控HK-2细胞ABCG2表达中的作用.方法 以ELISA法检测收集的16例男性痛风患者组及16qk男性健康人群组血清雌二醇水平,尿酸由自动生化仪测得.分别以不同浓度雌二醇刺激HK-2细胞24 h或48 h,以实时荧光定量PCR(qPCR)法检测ABCG2表达.以雌二醇或雌二醇及PI3K/Akt抑制剂LY294002共同刺激HK-2细胞,qPCR法检测ABCG2表达,蛋白印迹法检测Akt、p-Ser473-Akt、p-Thr308-Akt及ABCG2表达变化.采用方差分析或t检验分析数据.结果 痛风患者组血尿酸水平[(547±18)μmol/L]明显高于健康人群[(344±12)μmol/L](t=-5.395,P＜0.01).痛风患者组雌二醇水平[(45±6) μmol/L]较健康人群组[(100±8) μmol/L]明显降低(t=9.375,P＜0.01).10-4 mol/L组雌二醇刺激HK-2细胞24h或48 h后,ABCG2 mRNA水平均升高(t=3.168,t=3.990;P＜0.01).雌二醇及PI3K/Akt抑制剂LY294002共同干预HK-2细胞组,较单以雌二醇干预组,ABCG2、p-Ser473-Akt及p-Thr308-Akt表达下降,Akt表达差异无统计学意义.结论 雌二醇与血尿酸水平有明显的相关性,雌二醇可通过激活PI3K/Akt通路发挥上调人肾小管上皮细胞ABCG2表达的作用.

abstractsObjective To compare the serum estradiol level of gouty patients with healthy controls and to investigate whether estradiol upregulatesthe expression of the uric acid transporter ATP-binding cassette superfamily member 2 (ABCG2) of human renal tubular epithelial cells (HK-2)..Methods Serum of 16 male gout patients and 16 male healthy controls and their estradiol level were assessed with ELISA.The HK-2 cells wer cultured with different concentrations of estradiol for 24 hours or 48 hours.mRNA expression of ABCG2 was assessed by quantitative polymerase chain reaction (qPCR).HK-2 cells were cultured with estradiol or estradiol and inhibitor of PI3K/Akt pathway LY294002.mRNA expression of ABCG2 was assessed by qPCR,while the Akt,p-Ser473-Akt,p-Thr308-Akt and ABCG2 expression were investigated by Western blot.Data was analyzed using either the one-way analysis of variance or the t test.Results The level of serum uric acid in gout patients was [(547±18) μmol/L],which was significantly higher than that of healthy controls [(344±12) μmol/L),t=-5.395,P＜0.01].The level of estradiol in gout patients was (45±6) μmol/L,which was significantly lower than that of healthy controls [(100±8) μmol/L,t=9.375,P＜0.01].The mRNA expression of ABCG2 of 10-4 mol/L estradiol group was elevated after 24 hours or 48 hours (t=3.168,t=3.990;P＜0.01).In the group of co-stimulation withestradiol and LY294002,the ABCG2,p-Ser473-Akt and p-Thr308-Akt expression were down regulated compared to the estradiol group.Conclusion There is a significant correlation between serum estradiol and uric acid.Estradiol inducesthe expression of ABCG2 of HK-2 cells by activating PI3K/Akt pathway.