Effects of areca nut extracts on expression of adhesion molecules and degranulation in human polymorphonuclear neutrophils

• Main Authors: Hui-Wen Lee, 李惠文
• Other Authors: Shan-Ling Hung
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/54996032609734434023

碩士 === 國立陽明大學 === 口腔生物研究所 === 97 === Areca quid chewing is associated with a high prevalence of oral cancer, oral submucous fibrosis and periodontal disease. Aberrations of polymorphonuclear neutrophils (PMNs) linked with periodontal diseases have been reported. PMNs utilize adhesion molecules to help their rolling on endothelial cells of blood vessels, and the subsequent transmigration towards inflammatory foci. PMNs eliminate of foreign microorganisms through phagocytosis, degranulation and generation of reative oxygen species. Areca nut extracts (ANEs) enhanced the chemotaxis, adhesive ability to endothelial cells and the concentration of intracellular calcium. The purpose of the study was to investigate the effects of ANEs on expression of adhesion molecules and degranulation in PMNs. PMNs isolated from peripheral blood of heathy volunteers were resuspended in HBSS/Ca2+ buffer, and treated with various concentrations of ANEs for 30 minutes. The effects of ANE on size, granularity, and cell viability of PMNs were determined using flow cytometer after propidium iodide (PI) staining. In addition, the expression of adhesion molecules (selectin family : L-selectin, PSGL-1; β2 integrin: Mac-1 and LFA-1) and the surface membrane protein expression of primary granule marker (CD63), sencodary granule marker(CD66b) and secretory vesicle marker (CD45) was measured using immunofluorescence staining. The intracellular calcium chelator (BAPTA-AM) was also used to examine the effects of ANE on degranulation of primary granule in PMNs. The results showed that treatment with ANEs for 30 minutes, the size and granularity of PMNs were increased when compared with the control. The viability of PMNs was decreased to 85.70 %± 1.45 % when 25 μg/ml ANEs was used. Compared with non-treated PMNs, the expression of adhesion molecules (L-selectin, PSGL-1, Mac-1 and LFA-1) was decreased as measured by flow cytometry. When treasted with ANEs, the expression of primary granule marker (CD63) and secondary granule marker (CD66b) was increased, but the secretory vesicle marker (CD45) was decreased. Besides, after pretreated with the intracellular calcium chelator, BAPTA-AM, the inducing effect of ANE on surface expression of CD63 was inhibited. In conclusion, the results in this study showed that ANEs inhibited the expression of adhesion molecules, but induced primary and sencodary granules degranulation. The inducing effects in primary granules degranulation of ANEs may be due to the increased level of intracellular calcium. Taken together, the results of this study indicated that the influences of ANEs on the oral heath and inflammation may be through the regulation of adhesion molecules and degranulation in PMNs.