Radiology Department, Xianning Central Hospital, The First Affiliated Hospital of Hubei University of Science and Technology, Xianning, Hubei 437100, P.R. China.
Mol Med Rep. 2018 Jan;17(1):1205-1211. doi: 10.3892/mmr.2017.7975. Epub 2017 Nov 6.
Hyperglycemia mediated endothelial cells (ECs) injury is closely associated with diabetic vascular complications. It was revealed that DJ‑1 possesses cellular protective effects by suppressing oxidative stress. The present study aimed to investigate the beneficial effects of DJ‑1 on high glucose (HG)‑induced human umbilical vein endothelial cell (HUVEC) injury and to elucidate its underlying mechanisms. HUVECs were incubated under 5.5 mM (control group) or 25 mM D‑glucose (HG group) and then transfected with recombinant adenoviral vectors to overexpression of DJ‑1. Cell proliferation and apoptosis were measured using the EdU incorporation assay and flow cytometry with Annexin V-FITC/propidium iodide double staining, respectively. Apoptotic‑related proteins were determined using western blot analysis. Reactive oxygen species (ROS) production, lactate dehydrogenase (LDH) and nitric oxide (NO) levels, the content of malondialdehyde (MDA), and the activities of superoxide dismutase (SOD) were measured. Results demonstrated that overexpression of DJ‑1 promoted cell proliferation and inhibited HUVECs apoptosis stimulated by HG. DJ‑1 also suppressed the HG‑induced reduction in the Bcl‑2/Bax ratio and HG activated ROS generation in HUVECs. Furthermore, HG significantly increased the levels of LDH and MDA, and reduced the level of SOD; however, these effects were reversed by Ad‑DJ‑1 transfection. Furthermore, the cellular protective effect of overexpression of DJ‑1 enhanced p‑Akt/Akt ratio, eNOS activation and NO production, and these trends were partially reversed by a phosphatidylinositol‑4,5‑bisphosphate 3‑kinase (PI3K) inhibitor (LY294002). Taken together, the present study highlighted the involvement of DJ‑1 in HG‑related EC injury and identified that DJ‑1 exerts a cellular protective effect in HUVECs exposed to HG induced oxidative stress via activation of the PI3K/Akt‑eNOS signaling pathway.
高血糖介导的内皮细胞（EC）损伤与糖尿病血管并发症密切相关。研究表明 DJ-1 通过抑制氧化应激具有细胞保护作用。本研究旨在探讨 DJ-1 对高葡萄糖（HG）诱导的人脐静脉内皮细胞（HUVEC）损伤的有益作用，并阐明其潜在机制。将 HUVEC 在 5.5mM（对照组）或 25mM D-葡萄糖（HG 组）下孵育，然后用重组腺病毒载体转染以过表达 DJ-1。使用 EdU 掺入测定法和 Annexin V-FITC/碘化丙啶双染色流式细胞术分别测量细胞增殖和细胞凋亡。使用 Western blot 分析测定凋亡相关蛋白。测量活性氧（ROS）产生、乳酸脱氢酶（LDH）和一氧化氮（NO）水平、丙二醛（MDA）含量以及超氧化物歧化酶（SOD）活性。结果表明，过表达 DJ-1 促进了 HG 刺激的 HUVEC 增殖并抑制了细胞凋亡。DJ-1 还抑制了 HG 诱导的 HUVEC 中 Bcl-2/Bax 比例降低和 ROS 生成增加。此外，HG 显著增加了 LDH 和 MDA 的水平，降低了 SOD 的水平；然而，这些作用被 Ad-DJ-1 转染逆转。此外，过表达 DJ-1 的细胞保护作用增强了 p-Akt/Akt 比值、eNOS 激活和 NO 产生，这些趋势被磷脂酰肌醇-4,5-二磷酸 3-激酶（PI3K）抑制剂（LY294002）部分逆转。综上所述，本研究强调了 DJ-1 参与 HG 相关 EC 损伤，并确定 DJ-1 通过激活 PI3K/Akt-eNOS 信号通路在暴露于 HG 诱导的氧化应激的 HUVEC 中发挥细胞保护作用。
