Necroptosis mediates high glucose-induced injury in human umbilical vein endothelial cells |
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Authors: | LIN Jia-qiong CHEN Mei-ji GUO Rui-xian ZHANG Wei-jie ZHI Xi-mei DENG Hai-ou XU Ling LI Ying-lan WU Wen |
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Institution: | 1. Southern Medical University, Guangzhou 510515, China;
2. Department of Endocrinology, East Ward, Guangdong Geriatric Institute, Guangdong Academy of Medical Sciences, Guangdong General Hospital, Guangzhou 510080, China;
3. Department of Pediatrics, Huangpu Division of The First Affiliated Hospital of Sun Yat-sen University, Guangzhou 510700, China;
4. Department of Physiology, Zhongshan School of Medicine, Sun Yat-sen University, Guangzhou 510080, China |
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Abstract: | AIM: To explore whether necroptosis contributes to the high glucose (HG)-induced damage in human umbilical vein endothelial cells (HUVECs). METHODS: The protein levels of receptor-interacting protein 3 (RIP3) and cleaved caspase-3 were detected by Western blot. The intracellular levels of reactive oxygen species (ROS) were determined by DCFH-DA staining followed by photofluorography. Mitochondrial membrane potential (MMP) was measured by rhodamine 123 staining followed by photofluorography. RESULTS: Treatment of HUVECs with HG at different concentrations (10, 20 and 40 mmol/L glucose) for 24 h gradually enhanced the expression levels of RIP3. Treatment of HUVECs with HG (40 mmol/L glucose) for different time (3 h, 6 h, 9 h, 12 h and 24 h) also up-regulated the expression levels of RIP3, peaking at 9 h. Pretreatment of HUVECs with 20 μmol/L Z-VAD-FMK (an inhibitor of caspase) for 30 min before exposure to HG enhanced the expression level of RIP3. Pretreatment of HUVECs with 100 μmol/L necrostatin-1 (an inhi-bitor of necroptosis) for 1 h before exposure to HG alleviated the HG-induced injuries, such as a decrease in cell viability, an increase in ROS generation and dissipation of MMP, but up-regulated the protein level of cleaved caspase-3. CONCLUSION: Necroptosis mediates HG-induced injury in HUVECs. There is a negative interacting between necroptosis and apoptosis. |
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Keywords: | Necroptosis Apoptosis High glucose HUVECs Necrostatin-1 |
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