晚期糖基化终产物和同型半胱氨酸对内皮细胞的协同损伤作用

Co-injury effects of advanced Glycation end products and Homocysteine on endothelial cells

目的探讨两种尿毒症毒素晚期糖基化终产物(advanced glycation end products,AGE)和同型半胱氨酸(homocysteine,Hcy)在致血管内皮细胞损伤的过程中是否具有协同作用。方法不同浓度的AGE修饰牛血清白蛋白(AGE-BSA)和Hcy单独或联合作用于人脐静脉内皮细胞株ECV304细胞,检测各组上清液NO浓度和脂质过氧化终产物丙二醛(MDA)含量及细胞凋亡百分率。结果AGE+Hcy联合干预组的细胞凋亡率明显高于单独AGE和Hcy干预组(P<0.01),MDA含量也显著高于AGE干预组(P<0.05)和Hcy干预组(P<0.01),联合组干预组NO浓度则明显低于单独干预组(P<0.01)。结论AGE和Hcy均参与了血管内皮细胞的损伤,两者具有协同效应,上述效应可能与氧化应激有关,推测终末期肾病(ESRD)患者AGE和Hcy的升高可能是加速该人群动脉粥样硬化发生发展的重要因素。

[Objective] To investigate whether advanced Glycation end products （AGE） and Homocysteine （Hey） had co-effects on the injury of vascular endothelial cells. [Methods] ECV304 cell line, derived from human umbilical vein endothelial cells, were cultured in vitro with AGE modified bovine serum albumin （AGE-BSA） or Hey respectively and both together. Then 20 μl of 5 mg/mL MTT were added and the optical density （OD） at each concentration was determined in order to calculate the proliferation-inhibiting rate of cells. NO levels and MDA contents- results of lipid peroxidation in the supernatant were determined by using spectrophotometer. The ratio of apoptosis was assessed by Hoechst staining and flow cytometry techniques. [Results] The proliferation-inhibiting rates and apoptosis rates in AGE＋Hcy were both significantly higher than those in the AGE group （P 〈0.01） and Hcy group （P 〈0.01）. The MDA contents in AGE＋Hcy group were also significantly higher than those in the AGE group （P 〈0.05） and Hcy group （P 〈0.01）. But the NO levels in co-intervention group were significantly lower than those in solo intervention groups （P 〈0.01）. [Conclusion] Both AGE and Hcy can damage the vascular endothelial cells and they have co-injury effects on these cells, which may be related to oxidative stress and involved in the pathogenesis of atherosclerosis of the end stage renal disease （ESRD） patients.