蟲草萃取物於微神經膠細胞株中免疫調節作用之評估

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題名:	蟲草萃取物於微神經膠細胞株中免疫調節作用之評估
其他題名:	Immunomodulatory Effects of Cordyceps spp. in BV-2 Microglial cells
作者:	鐘婉予 Chung, Wan-Yu
貢獻者:	盧錫祺 Lu, Hsi-Chi 東海大學食品科學系
關鍵詞:	蟲草;微神經膠細胞;前列腺素;一氧化氮 Cordyceps spp.;microglial cell;prostanoids;nitric oxide
日期:	2006
上傳時間:	2011-04-13T08:18:14Z (UTC)
摘要:	腦部的發炎反應與多種神經退化性疾病相關。雖然分子機制未明，但是目前的研究已證實神經細胞的死亡和活化的微神經膠細胞有關。活化的微神經膠細胞會透過誘導型一氧化氮合成?(iNOS)和第二型環氧?(COX-2)產生大量一氧化氮(NO)和前列腺素E2 (PGE2)以及分泌細胞激素引發發炎反應，而過度的發炎反應會造成神經細胞的傷害、進而導致神經系統的異常和疾病。因此抑制發炎相關的因子是研究預防或治療發炎藥物的一個重要標的。冬蟲夏草Cordyceps sinensis 和蛹蟲草Cordyceps militaris 為已知具有抗腫瘤、抗氧化、治療腎炎及肝炎等活性之中草藥。研究中使用微神經膠細胞株BV-2做為評估模式，以冬蟲夏草及蛹蟲草液態發酵菌絲體萃取物之不同濃度溶於培養基中與細胞培養，加以LPS/IFN-r誘導細胞活化。從培養液中細胞釋出的一氧化氮和前列腺素E2與細胞激素IL-1b、IL-6和TNF-a的變化量，來探討蟲草萃取物對發炎相關因子生成量的作用。以及藉由西方墨點法與同步定量聚合?連鎖反應，分析誘導型一氧化氮合成?與環氧化?-2在蛋白質和基因層次的表現。結果顯示，蛹蟲草甲醇萃取物能抑制NO/iNOS和PGE2/COX-2這兩種發炎途徑，並且降低IL-1b、IL-6和TNF-a的釋放。冬蟲夏草乙醇萃取物的抗發炎效果與蛹蟲草甲醇萃取物相似，但在PGE2/COX-2的表現上有促進的效果。蛹蟲草水萃取物無明顯抗發炎作用，但可降低TNF-a生成。比較三種萃取物之抗發炎作用，以蛹蟲草甲醇萃取物最佳，其次為冬蟲夏草乙醇萃取物，最後為蛹蟲草水萃取物。由於不同萃取溶劑所得成分不相同，因此在調節免疫的途徑以及作用有異。透過不同發炎標記分子之評估，本研究探討蟲草萃取物之可能抗發炎反應，有助於建立中草藥與保健食品抑制神經系統發炎反應的測試平臺。 Inflammation in brain is associated with many neurodegenerative disorders. Though the molecular mechanism involved is not clear, evidences link neuron death to the activation of microglia. Through generation of nitric oxide and prostaglandin E2 by inducible nitric oxide synthase and cyclooxygenase-2, and secretion of cytokines, activated microglia promote inflammation responses, and may in turn lead to damage of neuron and dysfunction of nerve system when uncontrolled. Researches have targeted those inflammation pathways for inhibitors.Two related Chinese herbs, Cordyceps sinensis and C. militaris, have long been used for treating nephritis and hepartits and in recent studies demonstrated their anti-tumor and anti-oxidative activities. In this study, we evaluated the anti-inflammatory activity of fermented C. sinensis and C. militaris extracts in BV-2 microglia cells. After induction of inflammatory reaction by LPS/IFN-????production of ?NO, PGE E2, IL-1?, IL-6, and TNF-? were measured in cells pre-treated with extracts. Furthermore, the protein and mRNA expression levels of iNOS and COX-2 were also accessed by Western blot and Real-time RT-PCR. The methanol extracts of C. militaris inhibited NO/iNOS and PGE2/COX-2 pathways, and reduced the secretion of IL-1? IL-6 and TNF-?. The ethanol extract of C. sinensis showed similar anti-inflammatory effect, yet it increased the expression of PGE2/COX-2. No significant anti-inflammatory activity other than a reduction of TNF-??production could be observed under the treatment of the water extract of C. militaris.We suggest that the diverse compositions in extracts from different solvents might lead to distinctive regulation in inflammatory responses. Through the survey of different marker molecules, our results may help in establishing a screening system for herb or functional foods which might have anti-inflammatory activities in the nervous system.
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