Integrative Biomedical Research Unit (IBRU), Faculty of Allied Health Sciences, Naresuan University, Phitsanulok, 65000, Thailand.
Graduate Program in Biomedical Sciences, Faculty of Allied Health Sciences, Naresuan University, Phitsanulok, 65000, Thailand.
BMC Complement Med Ther. 2021 Feb 18;21(1):66. doi: 10.1186/s12906-021-03243-z.
High glucose (HG)-induced reactive oxygen species (ROS) overproduction impairs angiogenesis that is one pivotal factor of wound healing process. Angiogenesis impairment induces delayed wound healing, whereby it eventually leads to amputation in cases of poorly controlled diabetes with diabetic ulceration. Porcine placenta extract (PPE) is a natural waste product that comprises plenty of bioactive agents including growth factors and antioxidants. It was reported as an effective compound that prevents ROS generation. The goal of this study was to investigate the in vitro effect of PPE on HG-induced ROS-mediated angiogenesis impairment.
Primary endothelial cells (HUVECs) and endothelial cell line (EA.hy926) were treated with HG in the presence of PPE. The endothelial cells (ECs) viability, intracellular ROS generation, migration, and angiogenesis were determined by MTT assay, DCFDA reagent, wound healing assay, and tube formation assay, respectively. Additionally, the molecular mechanism of PPE on HG-induced angiogenesis impairment was investigated by Western blot. The angiogenic growth factor secretion was also investigated by the sandwich ELISA technique.
HG in the presence of PPE significantly decreased intracellular ROS overproduction compared to HG alone. HG in the presence of PPE significantly increased ECs viability, migration, and angiogenesis compared to HG alone by showing recovery of PI3K/Akt/ERK1/2 activation. HG in the presence of PPE also decreased ECs apoptosis compared to HG alone by decreasing p53/Bax/cleaved caspase 9/cleaved caspase 3 levels and increasing Bcl 2 level.
PPE attenuated HG-induced intracellular ROS overproduction that improved ECs viability, proliferation, migration, and angiogenesis by showing recovery of PI3K/Akt/ERK1/2 activation and inhibition of ECs apoptosis. This study suggests PPE ameliorated HG-induced ROS-mediated angiogenesis impairment, whereby it potentially provides an alternative treatment for diabetic wounds.
高糖（HG）诱导的活性氧（ROS）过度产生会损害血管生成，这是伤口愈合过程的一个关键因素。血管生成受损会导致伤口愈合延迟，最终导致糖尿病患者的溃疡处出现坏疽而截肢。猪胎盘提取物（PPE）是一种天然的废物，其中包含大量生物活性物质，包括生长因子和抗氧化剂。它被报道为一种有效的化合物，可以防止 ROS 的产生。本研究的目的是研究 PPE 在体外对 HG 诱导的 ROS 介导的血管生成损伤的作用。
用 HG 处理原代内皮细胞（HUVEC）和内皮细胞系（EA.hy926），并在 PPE 存在的情况下处理。通过 MTT 测定法、DCFDA 试剂、划痕愈合试验和管形成试验分别测定内皮细胞（EC）活力、细胞内 ROS 生成、迁移和血管生成。此外，通过 Western blot 研究 PPE 对 HG 诱导的血管生成损伤的分子机制。还通过夹心 ELISA 技术研究了 PPE 对 HG 诱导的血管生成生长因子分泌的影响。
与 HG 单独处理相比，PPE 存在时 HG 显著降低了细胞内 ROS 的过度产生。与 HG 单独处理相比，PPE 存在时 HG 显著增加了 ECs 的活力、迁移和血管生成，表现为 PI3K/Akt/ERK1/2 激活的恢复。与 HG 单独处理相比，PPE 存在时 HG 还通过降低 p53/Bax/cleaved caspase 9/cleaved caspase 3 水平和增加 Bcl-2 水平降低了 ECs 的凋亡。
PPE 减轻了 HG 诱导的细胞内 ROS 过度产生，通过恢复 PI3K/Akt/ERK1/2 激活和抑制 ECs 凋亡，提高了 ECs 的活力、增殖、迁移和血管生成。本研究表明，PPE 改善了 HG 诱导的 ROS 介导的血管生成损伤，为糖尿病伤口提供了一种潜在的治疗方法。
