論文名稱:

檳榔嚼塊成分對於人類中性白血球基質金屬蛋白酶-9及分裂原活化蛋白激酶的影響

 

Effects of components of areca quid on matrix metalloproteinase-9 and

 

mitogen-activated protein kinases in human polymorphonuclear neutrophils

研究生:

劉衛駿  Wei-Chun Liu

 

(以作者名查詢陽明大學館藏系)

 

(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:

39317004

          學年度:

94

          語文別:

中文

          出版年:

95

關鍵字:

中性白血球  PMN, neutrophil

 

基質金屬蛋白-9  matrix metalloproteinase-9, MMP-9

 

分裂原活化蛋白激  MAPK

 

檳榔  areca

 

檳榔素  arecoline

 

黃樟素  sarole

 

檳榔萃取物  ANE

 

丁香  eugenol

全文說明:

(本論文 20110824 對校內公開)

 

電子全文

        論文頁數:

87

摘要:

在台灣,有超過兩百萬人有嚼食檳榔的習慣。檳榔嚼塊成分會抑制人類中性白血球(polymorphonuclear neutrophils)的殺菌力。中性白血球是對抗病原的第一道防線,活化後會釋放顆粒的內含物,其中包含了基質金屬蛋白酶-9 (matrix metalloproteinase-9, MMP-9),具有降解細胞外基質(extracellular matrix)的功能,而分裂原活化蛋白激酶(mitogen-activated protein kinase, MAPK)可以調控MMP-9的釋放。本研究探討檳榔嚼塊成分是否影響中性白血球釋放MMP-9,並探討對MAPK活化的影響。利用健康自願者的周邊血液純化出嗜中性白血球,以不同檳榔嚼塊成分:檳榔萃取物(areca nut extract, ANE)、檳榔素(arecoline)、黃樟素(safrole)及丁香(eugenol)來處理,分析細胞存活度(cell viability)MMP-9活性、MMP-9蛋白質的表現及p38, ERK, JNK (MAPK)磷酸化的影響。研究結果顯示以10 µg/ml arecoline處理30分鐘並不影響中性白血球細胞存活度,以20 µg/ml ANE5 mM safrole3 mM eugenol處理30分鐘,50%的細胞會死亡。Arecoline抑制MMP-9的活性,而ANEsafroleeugenol則使MMP-9活性上升;隨著時間的增加,arecolinesafroleeugenol處理細胞存活度下降,同時arecoline抑制MMP-9活性,而safroleeugenol則會增加MMP-9的活性。此外,以不同濃度的檳榔嚼塊成分來處理中性白血球三十分鐘後,arecoline會使細胞內MMP-9蛋白質表現量上升,ANE則會下降,而safroleeugenol不影響細胞內MMP-9蛋白質表現。檳榔嚼塊成分處理30分鐘並不影響p38磷酸化的情形,但arecoline會使ERK磷酸化減低,而safrole則是ERK磷酸化增加,ANEeugenol則是使JNK磷酸化情形增加。本研究顯示檳榔嚼塊成分會影響中性白血球釋放MMP-9,並影響MAPK的磷酸化。

 

There are more than two million people chewing AQ in Taiwan. The components of AQ can inhibit the bactericidal activity of human polymorphonuclear neutrophils (PMNs). PMNs are the first line of defense to resist pathogen. After activation, PMNs will release enzymes, including matrix metalloproteinase-9 (MMP-9) which can degrade extracellular matrix from granules. The release of MMP-9 can be regulated by mitogen-activated protein kinase (MAPK). This study was to investigate the effects of the components of AQ on release of MMP-9 and activation of MAPK in PMNs. PMNs isolated from peripheral blood of healthy volunteers were treated with different components of AQ, including arca nut extract (ANE), arecoline, safrole and eugenol. Then analysed the viability of PMNs, activity of extracellular MMP-9,  intracellular production of MMP-9 protein and phosphorylation of p38, ERK and JNK. The results showed that treatment with 10 µg/ml arecoline for 30 minutes did not affect the viability of PMNs. However, the viability of PMNs was approximately 50% when 20 µg/ml of ANE, 5 mM of safrole or 3 mM eugenol was used for 30 minutes. The activity of MMP-9 was inhibited by arecoline but enhanced by ANE, safrole or eugenol. Time-dependent decrease in the viability was observed following the treatment of arecoline, safrole or eugenol, the activity of MMP-9 was inhibited by arecoline but enhanced by safrole or eugenol. Incubation of PMNs with various concentrations of these components for 30 minutes, the intracellular production of MMP-9 protein was enhanced by arecoline but inhibited by ANE. However, the intracellular production of MMP-9 protein was not affected by safrole or eugenol. The phosphorylation of p38 was not affected in PMNs by these components after treatment for 30 minutes. However, the phosphorylation of ERK was inhibited by arecoline but enhanced by safrole. In addition, the phosphorylation of JNK was enhanced by ANE and eugenol. In comclusion, the components of AQ regulated the release of MMP-9 and phosphorylation of MAPK in PMNs.

        發表著作

Hung S-L, Lin Y-J, Chien E-J, Liu W-G, Chang H-W, Liu T-Y, Chen Y-T (corresponding author). 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)