GLNVA抑制內毒素在鼠科小神經膠細胞株BV-2和巨噬細胞株RAW264.7引起發炎媒介物之表現

GLNVA REDUCED LPS-INDUCED INFLAMMAORY MEDIATORS IN MOUSE MICROGLIAL CELL BV-2 AND MACROPHAGE RAW 264.7

汪昊緯(Hao-Wei Uang)

指導教授 ： 羅怡卿

高雄醫學大學/醫學院/醫學研究所/碩士(2005年)

摘要

Glyceryl Nonivamide(GLNVA)是一個無辛辣味的辣椒素(Capsaicin)衍生物。GLNVA已被證實過的藥理作用，包括：降血壓作用，增加心血管之感覺神經元的calcitonin gene related peptide (CGRP)和P物質(substance P)釋放作用，並引起相關之鉀離子通道的打開作用，抑制腦下垂體之鉀離子通道，增加腎功能和預防蜘蛛膜下腔出血的腦血管痙攣等作用。此篇論文將著重於探討GLNVA的抗發炎作用。研究結果指出在小鼠的巨噬細胞株RAW264.7 和小神經膠細胞(microglia)株BV-2中，GLNVA能有效的抑制由革蘭氏陰性菌的脂多醣體(Lipopolysaccharide；LPS)所引發的發炎反應。Nitric Oxide(NO)的測定證明GLNVA能夠抑制LPS所引發的NO釋放；西方點墨法顯示GLNVA能夠濃度相關性(1、10、100 ?嵱)抑制以LPS處理過後的iNOS表現及COX-2表現，且能抑制I?羠?恁BI?羠?狾bLPS處理後之分解，並且在以LPS處理後，抑制ERK的磷酸化現象。另以ELISA測量發炎相關的細胞激素(cytokines)，發現GLNVA也能有效的抑制以LPS分別處理後TNF-?恁BIL-1?牷BPGE2的釋放，且也能夠使抗發炎的細胞激素IL-10釋放增加。依上面的結果，我們可推測GLNVA具有抗發炎作用，而其抗發炎機轉可能和NF-?羠的路徑有關，而根據EMSA的實驗，顯示GLNVA能抑制LPS引起的NF-?羠移進核內而轉錄的能力。因在BV-2中內毒素無法引起ERK1/2的活化，所以GLNVA的抗發炎能力在兩細胞中是不同的。總結來說GLNVA能夠抑制因LPS引發的發炎反應，其作用機轉包括抑制LPS引起的：NO/iNOS、COX-2、TNF-?恁BIL-1?牷BPGE2的釋放，I?羠?恁BI?羠?猁漱戲恁AERK的磷酸化作用，及NF-?羠與DNA結合的能力。

關鍵字

內毒素 ； GLNVA ； 小神經膠細胞 ； 巨噬細胞 ； 發炎媒介物 ； NF-kappaB

並列摘要

Glyceryl Nonivamide (GLNVA) was a nonpungent and antinociceptive capsaicin derivative. GLNVA has reported for several pharmacological effects including antihypertension, calcitonin gene related peptide (CGRP) and substance P releasing, enhancement of renal function and prevention subarachnoid hemorrhage-induced cerebral vasospasm. In this study, the effects of GLNVA on protein expressions related to inflammation were investigated in murine macrophage RAW264.7 and immortalized microglia cell line BV-2. Pretreatment of cells with GLNVA diminished lipopolysaccharide (LPS)-induced nitric oxide (NO) production in a concentration-dependent manner. GLNVA inhibition of LPS-induced NO production accompanied by suppression of inducible NO synthase (iNOS) expression. ERK1/2 phosphorylation and I?羠??/I?羠?? degradation were also inhibited by GLNVA in RAW264.7 and BV-2. Although COX-2 was slightly inhibited by GLNVA, LPS-induced PGE2 level was lowered by GLNVA in dose -dependent manners. The potency for ERK inhibition and I?羠??/I?羠?? degradation displayed relative correlations with their inhibitory effects on translocation into nucleus and DNA binding affinity of NF-?羠 and NO production. GLNVA also decreases pro-inflammatory cytokines-TNF-?? and IL-1β release and increases anti-inflammatory cytokine-IL-10 production. LPS could not induce ERK1/2 phosphorylation in BV-2 and it might be the reason that there are different effects of GLNVA in LPS-activated two cell lines. Taken together, GLNVA exerts its anti-inflammatory effect by inhibiting LPS-induced pro-inflammatory mediators, and thus could be used as a potential agent for anti-inflammation.

並列關鍵字

LPS ； microglia ； macrophage ； NF-kappaB ； inflammatory mediators ； GLNVA

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