The Membrane Fluidity and The Function of Neutrophils from Periodontitis Patients.

为探讨牙周病患者中性粒细胞膜流动性和细胞功能的变化，本研究从牙周病患者的牙周炎中分离得到中性粒细胞胞外小泡，分别从牙周炎和牙周病患者的牙周炎中提取脂多糖，从粘性放线菌T14V中分离纯化两亲性放线菌。健康志愿者中性粒细胞在ECV存在和不存在的情况下，其超氧化物生成、吞噬功能和膜流动性均受到明显的抑制，吞噬功能随ECV浓度的增加而略有下降，且ECV浓度的增加对膜流动性有轻微的抑制作用，而对深层膜流动性无明显影响。ACA…的作用对血管内皮细胞的膜流动性和通透性进行了体外检测。结果表明，细胞膜流动性增加，内质乳酸脱氢酶在培养液中的释放呈剂量和时间依赖性。观察糖尿病晚期牙周炎患者中性粒细胞超氧化物歧化、趋化性和膜流动性的变化。糖尿病患者中性粒细胞产生的超氧化物歧化能力明显高于健康对照组。趋化性和膜流动性略有下降。与健康对照组相比，DM患者中性粒细胞产生的超氧阴离子交换能力明显增强，提示牙周袋或牙周组织中的牙周病原体ECV或LPS可改变中性粒细胞和内皮细胞的膜流动性和细胞功能，提示牙周疾病的发生和发展。较少

The purpose of this research project was to investigate the membrane fluidity and the cell function of neutrophils from the patients with periodontal diseases.The extracellular vesicle (ECV) was obtained from P.gingivalis ATCC 33277, the lipopolysaccharides (LPS) were extracted from P.gingivalis 381 and P.intermedia ATCC 25611, and Actinomyces amphiphile (AcA) was purified from A.viscosus T14V.The following studies were performed on the obtained ECV or LPS.1. Superoxide production, phagocytosis and membrane fluidity were subjected by the neutrophils from healthy volunteers in the presence and absence of ECV.The superoxide production showed the significant suppression and the phagocytosis were slightly decreased with the increasing concentrations of ECV.Furthermore, the increasing concentrations of ECV slightly depressed the membrane fluifity at a shallow portion of lipid bilayr, whereas, showed no effects on the membrane fluidity at a deep portion of lipid bilayr.2. The effects of AcA … More on the membrane fluidity and the permeability of endothelial cells were evaluated in vitro. The results showed the increase of membrane fluidity on the cells and the release of endoplasmic LDH in the culture medium dose-and time-dependently.3. Superoxide production, chemotaxis and membrane fluidity were subjected by the neutrophils from Diabetes Mellitus (DM) patients with advanced periodontitis. The superoxide production by neutrophils from DM patients was significantly higher than those from healthy control. The chemotaxis and the membrane fluidity were slightly decreased. The superoxide production by neutrophils from DM patients was highly stimulated by the LPS compared to those from healthy control.These results indicated that the membrane fluidity and the cell function of neutrophils and endothelial cells were altered by the ECV or LPS from periodontal pathogens in the periodontal pockets or periodontal tissues, suggesting the initiation and the progression of periodontal diseases. Less