臺灣博碩士論文加值系統

從靈芝純化出來的多醣體萃取物過去已發現具有抗發炎及免疫調控的能力。過去的研究發現靈芝多醣體萃取物刺激老鼠脾臟細胞可產生一種稱為顆粒性白血球群落刺激因子的細胞激素。然而過去對於靈芝多醣體萃取物刺激產生顆粒性白血球群落刺激因子的機制尚未清楚。在我們的研究中，我們利用人類細胞株THP-1巨噬細胞及人類巨噬細胞，作為研究靈芝多醣體刺激產生顆粒性白血球群落刺激因子之訊息傳導途徑的材料。我們發現在在人類巨噬細胞中，靈芝多醣體萃取物可以透過類鐸受體2和4去調控顆粒性白血球群落刺激因子的產生。但在人類細胞株THP-1巨噬細胞中，只透過類鐸受體2去調控顆粒性白血球群落刺激因子的產生。接下來我們想要了解靈芝多醣體是透過何種蛋白化激�‘h調控顆粒性白血球群落刺激因子的產生。我們發現在人類細胞株THP-1巨噬細胞及人類巨噬細胞中，當使用PI-PLC、ERK的抑制劑時皆可抑制由靈芝多醣體萃取物刺激產生顆粒性白血球群落刺激因子的分泌量。而使用PKCα、PI3K抑制劑時，只影響人類巨噬細胞而不影響人類細胞株THP-1巨噬細胞產生顆粒性白血球群落刺激因子的分泌量。因此，我們發現靈芝多醣體萃取物可刺激人類巨噬細胞產生顆粒性白血球群落刺激因子，藉此特性可具有潛力用來當作免疫調控之藥物。

Extract of Reishi polysaccharides (EORP or fraction 3) from Ganoderma lucidum has been demonstrated to possess immuno-modulating effects. Previous study has shown that EORP could induce granulocyte colony stimulating factor (G-CSF) production in mouse spleen cells. However, the mechanism of EORP-induced G-CSF production is still not clear. In our study, we used human THP-1 macrophages and human primary macrophages to study the signal transduction pathway of EORP-induced G-CSF production. We found that EORP regulates G-CSF production through toll-like receptor 2 and 4 in human primary macrophages; but through toll-like receptor 2 in human THP-1 macrophages. Next we want to know which protein kinases participating in EORP-induced G-CSF production. We found out G-CSF production were inhibited when using inhibitor of phosphoinositide-specific phospholipase C and ERK both in humanTHP-1 macrophages and human primary macrophages. However, inhibitors of PKCα and PI3K had effect only in human primary macrophages but not in human THP-1 macrophages. As a conclusion, EORP could regulate G-CSF secretion in human macrophages and suggesting it acts as a pharmacological potential immune modulation agent.

中文摘要…………………………………………1
Abstract…………………………………………2
1.Introduction…………………………………3
2.Materials and Methods……………………7
2.1 Cells and cell culture
2.2 Culture of human primary macrophages
2.3 Materials
2.4 Preparation of EORP
2.5 G-CSF determinations
2.6 Western blot analysis
2.7 Statistical analysis
3.Results…………………………………………11
3.1 Extract of Reishi polysaccharides (EORP)
induces G-CSF secretion within human THP-1
macrophages and human primary macrophages
3.2 β (1→3)-D-glucan in EORP may play less role
in the G-CSF production
3.3 Toll-like receptors mediate signaling in EORP-induced
G-CSF secretion within human THP-1 macrophages and
human primary macrophages
3.4 EORP induces mitogen activated protein kinases (MAPKs)
phosphorylation in the regulation G-CSF production
within human THP-1 macrophages and human primary
macrophages
3.5 Phosphoinositide-specific phospholipase C regulate
G-CSF secretion within EORP-stimulated human THP-1
macrophages and human primary macrophages
3.6 Differential regulation of protein kinase C in EORP-
induced G-CSF secretion in human THP-1 macrophages and
human primary macrophages
3.7 Effect of PI3K inhibitor on EORP-induced G-CSF
secretion in THP-1 macrophages and human primary
macrophages
4.Discussion…………………………………………………17
5.References…………………………………………………23
6.Figure legends……………………………………………30

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