β-多醣体与TLR4之免疫功能.DOCVIP

PAGE PAGE 1 β-多醣體與TLR 4之免疫功能評估 The immunoregulatory effects of β-glucan and Toll-Like Receptor 4 function 研究生：白惠珠 Pai, Hui-Chu 指導教授：莊國賓 Chuang, Kuo-Pin 黃卓治 Huang, Tzou-Chi 【摘要】 敗血症(sepsis)大多為微生物感染造成無法控制的全身性發炎反應，包含大量血球細胞移動到組織及Toll-like receptors 4 (TLR4)的不正常調控，儘管予以廣泛性抗生素予以治療其所造成死亡率仍然很高。β-多醣體(β-glucan)的功效早己被廣為研究討論，並且已有多篇文獻報告顯示具有免疫調節的作用，可活化巨噬細胞及發炎物質的產生。探討敗血症誘導黏著分子Very Late Antigen-4 (VLA-4)產生與β-多醣體關係之研究資料發表不多。因此，使用脂多醣體(Lipopolysaccharide；LPS)刺激U-937人類單核球細胞株，並以流式細胞儀偵測細胞表面VLA-4黏著分子表現量。在實驗中發現，LPS促使黏著分子VLA-4表現量增加。再使用不同濃度的β-多醣體刺激U-937，隨著β-多醣體濃度增加而U-937表面黏著分子VLA-4表現量遞減。更進一步探討β-多醣體與LPS共同處理 U-937的狀況，確實在實驗數據顯示LPS所誘發之發炎反應可在β-多醣體共同反應下降低VLA-4表現量。另一方面，TLRs可以藉著先天免疫反應(innate immune system)而辨識病原體而引發免疫反應。LPS透過TLR4辨識而傳導發炎反應的訊息傳導路徑。在豬隻、小鼠、人類的比較上，不管在解剖構造或生理功能上豬隻比小鼠更相近於人。因而進一步選殖豬隻肺泡巨噬細胞(Alveolar macrophage)TLR4基因，目前已將豬隻TLR4 ECD質體DNA (porcine Toll-like receptor 4；pTLR4)轉染於Human embryo kidney 293 cells (HEK-293)，經由十二烷基硫酸鈉－聚丙烯醯胺膠體電泳法(sodium dodecyl sulfate-polyacrylamide gel electrophoresis；SDS）及西方墨點法(Western blotting)証實已得到穩定表現pTLR4 ECD蛋白質的細胞株。希望藉由此方面研究探討，提供一些關於β-多醣體與敗血症的相互關係，作為臨床上敗血症的相關治療參考。 關鍵字：脂多醣體、Toll-like receptor 4、β-多醣體、Dectin-1 【Abstract】 Sepsis occurs when microbes activate Toll-like receptors (TLRs) stimulating widespread inflammation. Uncontrolled activation of Lipopolysaccharide (LPS) induced mechanisms results in sepsis or systemic inflammatory response syndrome, including amount of blood cell migration into the tissue and unmoral expression of TLR4. The morbidity and mortality of sepsis remain high despite extensive treated with antibiotics. The immunostimulating properties of β-glucan have been known for many years. However, how these work between sepsis induced Very Late Antigen-4 (VLA-4) up-regulation and β-glucan to modulate macrophage response remains unclear. We treated human monocyte-like cell line U-937 with different concentration of β-glucan, the results indicated that the cell surface VLA-4 molecular were decreased by β-glucan in a dose depe