論文名稱:

檳榔萃取物調節周邊血液單核細胞功能之活化

 

Regulation of Areca Nut Extracts on Functional Activation of Peripheral Blood Mononuclear Cells

研究生:

岑枝梅  Jy-Mei Tsern

 

(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:

萬曉晴  Hsiao-Ching Wan

        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:

870H05

          學年度:

88

          語文別:

中文

          出版年:

89

關鍵字:

檳榔萃取物  Areca nut

 

周邊血液單核細胞  Peripheral Blood Mononuclear Cells; PBMC

        論文頁數:

56

摘要:

近年來嚼食檳榔已成為台灣地區相當普遍之習慣。流行病學研究指出,咀嚼檳榔與口腔黏膜下纖維化及口腔癌的發生有密切的關係。在動物模型和組織培養中,檳榔萃取物被證實可促進腫瘤生成和造成細胞傷亡。然而,其導致口腔疾病的致病機轉仍不十分清楚。在本研究中,我們從新的角度,探討檳榔與免疫細胞活化的關係。利用致死劑量以下的檳榔萃取物,刺激人體周邊血液單核細胞,來檢測其調節發炎細胞激素的功能,以及對牙齦纖維母細胞生長的影響。首先,將10-100 μg/ml老檳榔萃取液、20-200 μg/ml嫩檳榔萃取液或10-100 μg/ml arecoline與周邊血液單核細胞培養82448小時後,分別以RT-PCRELISA偵測細胞激素mRNA和蛋白質的表現。我們發現,老檳榔萃取物及嫩檳榔萃取物 (但非arecoline),均可刺激TNF-αIL-1β的分泌。其中老檳榔萃取物的調節作用較緩和而持久;嫩檳榔萃取物的影響則快起而短暫。同時,個體間對檳榔敏感性不同,所釋放的細胞激素的量也有相當的差異。另一方面,收集周邊血液單核細胞與50 μg/ml老檳榔萃取物或200 μg/ml嫩檳榔萃取物共同作用824小時的上清液,培養牙齦纖維母細胞,並利用BrdU標定新合成的DNA,以了解其對纖維母細胞增生的調控。結果顯示,在控制組中未受刺激的血液單核細胞培養液可促進纖維母細胞增生。但受檳榔萃取物刺激的單核細胞上清液,對細胞增生的影響卻不一致,無法遽下結論。總結而言,檳榔萃取物誘發周邊血液單核細胞產生TNF-αIL-1β,可能在口腔疾病之病理過程中調控組織反應。然而,其機制及生理意義,仍有待進一步探討證實。

   
 

The habit of betel quid chewing, very common in Taiwan, has long been associated with an increased risk of oral submucous fibrosis and oral cancer.  In animal models and some tissue culture studies, it was shown that areca nut or its chemical components can promote tumor formation and cause cell death.  However, the pathogenesis of betel quid related oral diseases is not fully understood yet.  In this study, we aimed to investigate the regulation of areca nut extract (ANE) on cytokine elaboration of peripheral blood mononuclear cells (PBMC), and its effect on gingival fibroblast proliferation.  First, freshly isolated PBMC were stimulated with various concentrations of ripeANE (10-100 μg/ml), tender ANE (20-200 μg/ml) or arecoline (10-100 μg/ml) for 8, 24, and 48 hours.  The mRNA and protein of proinflammatory cytokines were then detected by RT-PCR and ELISA separately.  We found that both ripe and tender ANE (but not arecoline) could stimulate the production of TNF-αand IL-1β.  Ripe ANE caused steady and persistent upregulation of cytokines, whereas the regulatory effect of tender ANE was abrupt and short-lived.  Moreover, the PBMC response to ripe or tender ANE stimulation varied among different donors with regard to their sensitivity and released cytokine level. Secondly, gingival fibroblasts were cultured with the condition medium from PBMC incubated with 50 μg/ml ripe ANE or 200 μg/ml tender ANE for 8 or 24 hours.  With BrdU labeling, the modulation of fibroblast proliferation was monitored.  The result showed that the culture supernatant from unstimulated PBMC could increase fibroblast proliferation, but the effect of conditional medium of ANE-stimulated PBMC was variable.

 

  In summary, areca nut extracts can induce PBMC elaborating TNF-αand IL-1β, which may play important modulatory roles in the process of disease development.  However, its mechanisms and biological significance remain to be further investigated.