School of Chinese Materia Medica, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, P.R. China.
Zhongshan Ophthalmic Center, Sun Yat‑Sen University, Guangzhou, Guangdong 510060, P.R. China.
Mol Med Rep. 2017 Nov;16(5):7745-7751. doi: 10.3892/mmr.2017.7571. Epub 2017 Sep 21.
It has been shown that oxidative damage and inflammation caused by hyperglycemia in endothelial cells are key factors triggering diabetic vascular complications. The aim of the present study was to investigate the antioxidant and anti‑inflammatory effects of Danhong Huayu Koufuye (DHK)‑medicated serum on high glucose (HG)‑induced injury in endothelial cells, and examine its underlying mechanisms. EA. hy926 cells were treated with normal glucose, HG, or HG with DHK‑medicated serum. Cell viability was assessed using the MTT method. Apoptosis was detected using flow cytometry. Intracellular reactive oxygen species (ROS) levels were measured using the 2',7'‑dichlorodihydrofluorescein method. Cell culture supernatant was collected for detecting the activities of glutathione peroxidase (GPx) and superoxide dismutase (SOD), and the levels of malondialdehyde (MDA). The protein expression levels of intercellular adhesion molecule‑1 (ICAM‑1), nuclear factor‑κB (NF‑κB), hypoxia‑inducible factor‑1α (HIF‑1α) and vascular endothelial growth factor (VEGF) were determined using western blot analysis. The results revealed that DHK‑medicated serum accelerated the proliferation and inhibited the apoptosis of cells treated with HG (P<0.01) in a dose‑dependent manner. Compared with the HG group, the high levels of ROS and MDA were significantly reduced by DHK‑medicated serum (P<0.01). A 10% concentration of DHK‑medicated serum increased the activities of SOD and GPx by 59.4 and 95.5%, respectively. The high protein expression levels of ICAM‑1, NF‑κB, VEGF and HIF‑1α were significantly ameliorated by DHK‑medicated serum (P<0.01, vs. HG group). These findings indicated that DHK‑medicated serum protected EA. hy926 cells from HG‑induced injury and apoptosis through antioxidation and anti‑inflammatory effects.
已经表明,高血糖引起的内皮细胞氧化损伤和炎症是触发糖尿病血管并发症的关键因素。本研究旨在探讨丹红化瘀口服液含药血清对高糖诱导的内皮细胞损伤的抗氧化和抗炎作用,并探讨其潜在机制。用正常葡萄糖、高葡萄糖(HG)或 HG 加丹红化瘀口服液含药血清处理 EA.hy926 细胞。用 MTT 法评估细胞活力。用流式细胞术检测细胞凋亡。用 2',7'-二氯二氢荧光素法测定细胞内活性氧(ROS)水平。收集细胞培养上清液,检测谷胱甘肽过氧化物酶(GPx)和超氧化物歧化酶(SOD)活性以及丙二醛(MDA)水平。用 Western blot 分析检测细胞间黏附分子-1(ICAM-1)、核因子-κB(NF-κB)、缺氧诱导因子-1α(HIF-1α)和血管内皮生长因子(VEGF)的蛋白表达水平。结果表明,丹红化瘀口服液含药血清呈剂量依赖性地促进 HG 处理的细胞增殖并抑制其凋亡(P<0.01)。与 HG 组相比,丹红化瘀口服液含药血清显著降低 ROS 和 MDA 水平(P<0.01)。丹红化瘀口服液含药血清浓度为 10%时,SOD 和 GPx 活性分别增加 59.4%和 95.5%。丹红化瘀口服液含药血清显著改善了 ICAM-1、NF-κB、VEGF 和 HIF-1α 的高表达水平(P<0.01,与 HG 组相比)。这些发现表明,丹红化瘀口服液含药血清通过抗氧化和抗炎作用保护 EA.hy926 细胞免受 HG 诱导的损伤和凋亡。