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Glutathione in gingival crevicular fluid and its relation to local antioxidant capacity in periodontal health and disease

机译:牙龈沟液中的谷胱甘肽及其与牙周健康和疾病中局部抗氧化能力的关系

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摘要

>Aims: To determine possible changes in gingival crevicular fluid (GCF) antioxidant defence in chronic adult periodontal disease and to investigate the nature of the local radical scavenging mechanisms, with particular reference to glutathione.>Methods: GCF and plasma were collected from patients with chronic periodontitis and age and sex matched control subjects (n = 10). Polymorphonuclear leucocytes (PMNLs) were prepared and gingival epithelial cells (GECs) were collected by conventional methods from periodontally healthy subjects. PMNL were stimulated with F-Met-Leu-Phe after cytochalasin B treatment. Enhanced chemiluminescence was used to determine the total antioxidant capacity and to investigate the activity of cell fractions and reducing agents. GCF concentrations of reduced (GSH) and oxidised (GSSG) glutathione were determined by high performance liquid chromatography.>Results: Plasma and GCF from patients contained lower mean (SD) total antioxidant capacity (501.8 (123) μM Teq/litre and 658.3 (392) μM Teq/litre, respectively) compared with controls (577.9 (99.8) and 1351.5 (861) μM Teq/litre, respectively). Antioxidant light recovery profiles for GCF demonstrated a stepped response, not seen in plasma, which was inhibited by N-ethylmaleimide. This response was also detected in the cytosolic fraction of GEC and anaerobically stimulated PMNL. Similar antioxidant profiles, inhibitable by N-ethylmaleimide, were obtained with cysteamine, cysteine, and GSH. Control GCF contained high mean (SD) concentrations of glutathione (GSH, 1899.8 (494.4)μM; GSSG, 256.8 (152.4)μM). GCF from patients with periodontitis contained significantly lower amounts of GSH (mean, 1183.1; SD, 580.3μM) and GSSG (mean, 150.1; SD, 44.9μM).>Conclusions: GSH values and total antioxidant capacity are reduced in chronic periodontal disease. The high concentrations of GSH present in GCF in health are similar to those found extracellularly in the lung and may represent an important antioxidant and anti-inflammatory defence strategy common to exposed epithelial surfaces.
机译:>目的:为了确定慢性成人牙周疾病中龈沟液(GCF)抗氧化防御的可能变化,并研究局部自由基清除机制的性质,特别是谷胱甘肽。>方法: 从慢性牙周炎患者,年龄和性别匹配的对照受试者(n = 10)中收集GCF和血浆。通过常规方法从牙周健康受试者中制备多形核白细胞(PMNL),并收集牙龈上皮细胞(GEC)。细胞松弛素B处理后,用F-Met-Leu-Phe刺激PMNL。增强的化学发光用于确定总抗氧化剂能力,并研究细胞级分和还原剂的活性。通过高效液相色谱法测定还原型(GSH)和氧化型(GSSG)谷胱甘肽的GCF浓度。>结果:来自患者的血浆和GCF的平均抗氧化剂总容量(SD)较低(501.8(123)μM Teq / L和658.3(392)μMTeq / L)与对照(分别为577.9(99.8)和1351.5(861)μMTeq / L)相比。 GCF的抗氧化剂光恢复曲线显示出阶跃响应,在血浆中未见,这被N-乙基马来酰亚胺抑制。在GEC和厌氧刺激的PMNL的胞质部分中也检测到了这种反应。用半胱胺,半胱氨酸和谷胱甘肽可获得类似的抗氧化剂,可被N-乙基马来酰亚胺抑制。对照GCF含有高浓度(SD)的谷胱甘肽(GSH,1899.8(494.4)μM; GSSG,256.8(152.4)μM)。牙周炎患者的GCF含量明显低于GSH(平均值为1183.1; SD,580.3μM)和GSSG(平均值为150.1; SD,44.9μM)。>结论: GSH值和总抗氧化能力为减少慢性牙周病。健康中GCF中高浓度的GSH与肺部细胞外发现的GSH相似,可能代表暴露于上皮表面的一种重要的抗氧化剂和抗炎防御策略。

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