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High Levels of Glucose Induced the Caspase-3/PARP Signaling Pathway, Leading to Apoptosis in Human Periodontal Ligament Fibroblasts

机译:高水平的葡萄糖诱导Caspase-3 / PARP信号通路,导致人牙周膜成纤维细胞凋亡

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摘要

Periodontitis is one of the main complications of diabetes mellitus, and much research has been conducted on their relationship. However, the mechanism by which high glucose levels induce damage of periodontal ligament fibroblasts is still unclear. In this study, we investigated the effects of high glucose levels on apoptosis in human periodontal ligament fibroblasts and the possible mechanisms involved. Human periodontal ligament fibroblasts were cultured in DMEM with normal glucose (5.5 mM) and high glucose (35 mM) levels for 6, 12, or 24 h. Apoptosis was studied by flow cytometry, caspase assays, fluorescent real-time PCR, and Western-blot analysis. The different durations of high glucose incubation induced a time-dependent increase of apoptosis and caspase-3 activity in cultured human periodontal ligament fibroblasts. In addition, concentrations of caspase-3 and its substrate PARP in cultured human periodontal ligament fibroblasts increased in a time-dependent manner. Furthermore, a caspase-3 inhibitor could prevent the high glucose-induced apoptosis in human periodontal ligament fibroblasts. These data indicate that high glucose induces a time- and caspase-3-dependent increase in apoptosis in cultured human periodontal ligament fibroblasts. These results elucidate the mechanism for the regulation of human periodontal ligament fibroblast apoptosis caused by high glucose.
机译:牙周炎是糖尿病的主要并发症之一,目前已对其关系进行了大量研究。然而,高葡萄糖水平诱导牙周膜成纤维细胞损伤的机制仍不清楚。在这项研究中,我们调查了高葡萄糖水平对人牙周膜成纤维细胞凋亡的影响及其可能的机制。将人牙周膜成纤维细胞在具有正常葡萄糖(5.5 mM)和高葡萄糖(35 mM)水平的DMEM中培养6、12或24 h。通过流式细胞术,胱天蛋白酶测定,荧光实时荧光定量PCR和蛋白质印迹分析研究细胞凋亡。高葡萄糖温育的不同持续时间在培养的人牙周膜成纤维细胞中诱导了凋亡和caspase-3活性的时间依赖性增加。另外,培养的人牙周膜成纤维细胞中caspase-3及其底物PARP的浓度以时间依赖性方式增加。此外,caspase-3抑制剂可以阻止高糖诱导的人牙周膜成纤维细胞凋亡。这些数据表明,高葡萄糖诱导培养的人牙周膜成纤维细胞凋亡的时间依赖性和caspase-3依赖性增加。这些结果阐明了高葡萄糖引起的调节人牙周膜成纤维细胞凋亡的机制。

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