AGEs培养的人肾小球系膜细胞中二氢叶酸还原酶及O_2~-水平

目的:观察晚期糖基化终末产物(AGEs)对人肾小球系膜细胞(HMCs)中二氢叶酸还原酶(DHFR)和超氧阴离子(O-2)水平的影响。方法:在HMCs中加入5.6mmol/L葡萄糖(正常对照组)和200mg/LAGEs(AGEs组),平行培养48h后,采用Western blotting检测两组DHFR蛋白含量,采用二氢乙啶(DHE)法检测两组O-2的水平。结果:与正常对照组比较,AGEs组DHFR蛋白表达显著下调(P<0.05),O-2水平显著上调(P<0.01)。结论:AGEs诱导HMCs产生的O-2可能通过下调DHFR蛋白水平影响四氢生物喋呤(BH4)的生物补救合成途径,导致BH4减少而加重糖尿病肾病(DN)的氧化应激。

Effect of AGEs on Expression of DHFR and the Ailiration of O2-

Objective: To investigate the effect of AGEs on the activation of superoxide anion (O2-) and expression of DHFR in human glomerular mesangial cells (HMCs). Method: HMCs were divided into two groups randomly, the normal control group and the experimental group,each of them was treated with glucose (5.6mmol/L) and AGEs (200mg/L ) for 48h respectively. The expression of DHFR was assessed by Western blotting. Dihydroethidine fluorescence was used to determine O2-. Results: Compared with the control group, the expression of DHFR was significantly decreased in HMCs under AGEs(P<0.05); the O2- level was significantly increased in HMCs under AGEs(P<0.01). Conclusion: AGEs induced the increasing of O2- level in cultured HMCs, and this O2- serves as a brake on DHFR generation inducing a prominent decrease in the rate of BH4 formation,which promoted the renal oxidative stress.