論文名稱:

檳榔子萃取液對於中性白血球花生四烯酸代謝途徑的影響

 

Effects of areca nut extracts on arachidonic acid pathway in human neutrophils

研究生:

林映潔  Ying-Jie Lin

 

(以作者名查詢陽明大學館藏系統)

 

(以作者名查詢全國圖書書目資訊網)

指導教授:

洪善鈴  Shan-Ling Hung

        學位類別:

碩士

        學校名稱:

國立陽明大學

系所名稱:

口腔生物研究所

            學號:

39217007

          學年度:

93

          語文別:

中文

          出版年:

94

關鍵字:

中性白血球  neutrophils

 

檳榔子萃取液  areca nut extracts

 

白三烯素  leukotriene

 

前列腺素  prostaglandin

全文說明:

(本論文未授權公開)

 

電子全文

        論文頁數:

96

摘要:

嚼食檳榔(areca quid, AQ)者比未嚼食檳榔者有較高的機率患牙周病。花生四烯酸(arachidonic acid)代謝途徑包含環氧合-2 (cyclooxygenase-2, COX-2)5-脂氧合酶(5-lipoxygenase, 5-LO)代謝途徑。在牙周病人的牙齦溝液(gingival crevicular fluid, GCF)中可發現比正常人高的白三烯素B4 (leukotriene B4, LTB4)與前列腺素E2 (prostaglandin E2, PGE2)含量。白三烯素B4的製造者主要為嗜中性白血球(neutrophils)白三烯素B4為活化5-脂氧合酶代謝途徑而產生,而5-脂氧合酶的活化與細胞內鈣離子及p38 mitogen-activated protein kinase (MAPK)磷酸化有關,前列腺素E2為環氧合酶-2代謝途徑的產物。本論文為探討檳榔嚼塊之主要成分檳榔子的萃取液(areca nut extract, ANE)對於中性白血球花生四烯酸代謝途徑的影響。以各種濃度的檳榔子萃取液處理中性白血球不同時間點後,利用酵素免疫分析法(enzyme immunoassay, EIA)分析上清液中白三烯素B4及前列腺素E2的含量。利用Fura-2螢光染劑測量檳榔子萃取液對於細胞內鈣離子濃度的影響。以西方墨點法觀察檳榔子萃取液對嗜中性白血球p38 MAPK磷酸化及環氧合-2蛋白質表現量的影響。研究結果顯示隨著檳榔子萃取液濃度的增加中性白血球產生白三烯素B4分泌量也跟著增加,而當處理時間增加時,白三烯素B4的分泌量則下降。而檳榔子萃取液濃度的增加或處理時間增長,嗜中性白血球產生前列腺素E2分泌量跟著上升。檳榔子萃取液可刺激嗜中性白血球細胞內鈣離子濃度增加。以檳榔子萃取液處理中性白血球0.5小時後,p38 MAPK磷酸化的情形增加。檳榔子萃取液處理中性白血球20小時後,環氧合-2蛋白質表現量增加。本論文研究結果顯示檳榔子萃取液能活化嗜中性白血球花生四烯酸代謝途徑,有助於了解嚼食檳榔與牙周疾病之間的關係。

   
 

Areca quid (AQ) chewers have a higher prevalence of periodontal diseases than non-chewers. There are two major metabolic pathways of arachidonic acid, the cyclooxygenase-2 (COX-2) pathway and the 5-lipoxygenase (5-LO) pathway. The amounts of leukotriene B4 (LTB4) and prostaglandin E2 (PGE2) in gingival crevicular fluid (GCF) were higher in the periodontal disease sites than the healthy sites from healthy control. Neutrophils are the major source of LTB4. LTB4 is synthesized via the 5-LO pathway. Activation of 5-LO pathway is regulated by intracellular calcium and phosphorylation of p38 mitogen-activated protein kinase (MAPK). PGE2 is a product of the COX-2 pathway. The propose of the study was to investigate the effects of areca nut extract (ANE), the main component of AQ, on arachidonic acid metabolic cascade in neutrophils. Neutrophils were treated with various concentrations of ANE for different incubation times. The amounts of LTB4 and PGE2 in culture supernatants were measured using enzyme immunoassay (EIA). The effects of ANE on intracellular calcium concentration were measured using fluorescent dye, Fura-2. The effects of ANE on phosphorylation of p38 MAPK and expression of COX-2 protein were examined using Western blotting technique. The results showed that ANE induced the generation of LTB4 in a dose-dependent manner. Time-dependent decrease in generation of LTB4 was observed following the treatment of ANE. ANE also induced secretion of PGE2 in a dose-and time-dependent manner. ANE caused a rapid elevation of cytosolic calcium concentration in neutrophils. The phosphorylation of p38 MAPK was induced in neutrophils after ANE treatment for 0.5 hour. After treating neutrophils with ANE for 20 hours, the expression of COX-2 was increased. In conclusion, the results indicated that ANE regulated the activation of arachidonic acid metabolic cascade in neutrophils. These data may help to explain the association of AQ chewing and periodontal diseases.

   

       發表著作:

Hung S-L, Lin Y-J, Chien E-J, Liu T-Y, Chen Y-T. Areca nut extracts activated secretion of leukotriene B4 and phosphorylation of p38 mitogen-activated protein kinase nand elevated intracellular calcium concentrations in human polymorphonuclear leukocytes. Journal of Periodontal Research 2006 (accepted).