Zhao Wenting, Wang Zhen, Sun Zewei, Wang Shuai, Wu Mingjie, Zheng Liangrong
Department of Cardiology, The First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310003, P.R. China.
Mol Med Rep. 2017 May;15(5):3048-3054. doi: 10.3892/mmr.2017.6417. Epub 2017 Mar 30.
Elevated blood glucose levels contribute to a series of complications in patients with diabetes mellitus, including chronic ulcers and accelerated atherosclerosis. Dysregulated endothelial migration induced by high glucose is important in vascular‑associated complications. In the present study, cluster of differentiation (CD)97/adhesion G protein‑coupled receptor 5 (ADGRE5), a member of the G protein‑coupled receptor protein family, stimulated angiogenesis was investigated to determine its role in cell models of diabetes mellitus using lentivirus‑mediated overexpression and siRNA transfection. The results revealed reduced expression in high glucose‑treated human umbilical vein endothelial cells and in the endothelium of diabetic mice. Among the three CD97 isoforms, the majority of the expression of CD97 (EGF1,2,5) in the endothelial cells was regulated by high levels of glucose. Using stable lentivirus‑mediated transfection and clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR‑associated protein 9 (Cas9) technology, the present study constructed CD97‑overexpression and CD97‑knockout endothelial cell lines. Migration assays showed that the lentivirus‑mediated overexpression of CD97/ADGRE5 improved the inhibition of high glucose‑induced endothelial cell migration. In addition, using cytoskeleton staining, it was found that CD97 promoted membrane ruffling and lamellipodia formation. Cell division cycle 42, a small GTP‑binding protein, and its downstream factor, actin‑related protein 2, were involved in CD97‑induced actin reorganization in endothelial cells. Additionally, the use of transcription factor filter plate assays revealed that the nuclear translocation of signal transducer and activator of transcription 1 stimulated by high glucose contributed to the inhibited transcription of CD97. In conclusion, the present study established that the overexpression of CD97 improved high glucose‑induced dysfunction of endothelial cell migration. These findings provide insight to assist in identifying therapeutic targets with potential to ameliorate certain vascular complications of diabetes.
血糖水平升高会导致糖尿病患者出现一系列并发症，包括慢性溃疡和动脉粥样硬化加速。高糖诱导的内皮细胞迁移失调在血管相关并发症中起重要作用。在本研究中，利用慢病毒介导的过表达和小干扰RNA转染，研究了G蛋白偶联受体蛋白家族成员分化簇（CD）97/黏附G蛋白偶联受体5（ADGRE5）刺激血管生成在糖尿病细胞模型中的作用。结果显示，高糖处理的人脐静脉内皮细胞和糖尿病小鼠的内皮中CD97表达降低。在三种CD97亚型中，内皮细胞中大多数CD97（EGF1,2,5）的表达受高水平葡萄糖调节。本研究利用稳定的慢病毒介导转染和规律成簇间隔短回文重复序列（CRISPR）/CRISPR相关蛋白9（Cas9）技术构建了CD97过表达和CD97敲除的内皮细胞系。迁移实验表明，慢病毒介导的CD97/ADGRE5过表达改善了高糖诱导的内皮细胞迁移抑制。此外，通过细胞骨架染色发现，CD97促进了膜皱褶和片状伪足的形成。小GTP结合蛋白细胞分裂周期42及其下游因子肌动蛋白相关蛋白2参与了CD97诱导的内皮细胞肌动蛋白重组。此外，转录因子滤板分析显示，高糖刺激的信号转导和转录激活因子1的核转位导致了CD97转录的抑制。总之，本研究证实CD97过表达改善了高糖诱导的内皮细胞迁移功能障碍。这些发现为识别可能改善糖尿病某些血管并发症的治疗靶点提供了思路。
