Li Yuan, Chang Ye, Ye Ning, Chen Yintao, Zhang Naijin, Sun Yingxian
Department of Cardiology, The First Hospital of China Medical University, Heping, Shenyang, Liaoning 110001, P.R. China.
Mol Med Rep. 2017 May;15(5):2673-2680. doi: 10.3892/mmr.2017.6314. Epub 2017 Mar 10.
Advanced glycation end products (AGEs) restrain the proliferation of endothelial cells, which is an important determinant of diabetic vasculopathy. Mitochondrial biogenesis serves an essential role in cellular adaptation and repair. The current study aimed to investigate alterations in mitochondrial energy metabolism in human umbilical vein endothelial cells (HUVECs) and the latent mechanism regulated by AGEs. The proliferation of cultured HUVECs stimulated with AGEs was detected using an MTT assay and a real‑time cell analyzer (RTCA). Mitochondrial energy metabolism was measured using a Seahorse metabolic flux analyzer. Mitochondrial membrane potential was detected under fluorescence microscopy following staining with tetraethylrhodamine and MitoTracker Red. Respiratory chain complexes I‑V were detected using western blotting. MTT and RTCA assays demonstrated that AGEs treatment significantly inhibited the viability and proliferation of HUVECs when compared with bovine serum albumin treatment. Results from the Seahorse metabolic flux analyzer indicated that mitochondrial aerobic respiration and glycolysis declined following AGEs treatment. In addition, mitochondrial membrane potential and the expression of mitochondrial respiration chain complexes I/II/III/IV/V notably decreased in the presence of AGEs. In conclusion, the results of the present study indicated that AGEs exhibited an inhibitory effect on the proliferation in HUVECs potentially by mediating the dysfunction of mitochondrial energy metabolism and glycolysis. This may provide a new consideration for therapeutic methods in diabetic vascular complications.
晚期糖基化终末产物（AGEs）抑制内皮细胞的增殖，而内皮细胞增殖是糖尿病血管病变的一个重要决定因素。线粒体生物合成在细胞适应和修复中起重要作用。本研究旨在探讨人脐静脉内皮细胞（HUVECs）中线粒体能量代谢的变化以及AGEs调控的潜在机制。使用MTT法和实时细胞分析仪（RTCA）检测用AGEs刺激的培养HUVECs的增殖。使用海马代谢通量分析仪测量线粒体能量代谢。用四乙基罗丹明和MitoTracker Red染色后，在荧光显微镜下检测线粒体膜电位。使用蛋白质印迹法检测呼吸链复合体I-V。MTT和RTCA分析表明，与牛血清白蛋白处理相比，AGEs处理显著抑制了HUVECs的活力和增殖。海马代谢通量分析仪的结果表明，AGEs处理后线粒体有氧呼吸和糖酵解下降。此外，在AGEs存在的情况下，线粒体膜电位以及线粒体呼吸链复合体I/II/III/IV/V的表达显著降低。总之，本研究结果表明，AGEs可能通过介导线粒体能量代谢和糖酵解功能障碍对HUVECs的增殖产生抑制作用。这可能为糖尿病血管并发症的治疗方法提供新的思路。
