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題名 | 蟾蜍靈對細胞發炎反應的效益評估及其作用機制和標的分子探討=Significance of Bufalin in Cellular Inflammatory Cascades |
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作者 | 吳鈺琳; Wu, Yuh-lin; |
期刊 | 中醫藥年報 |
出版日期 | 20141200 |
卷期 | 3 2014.12[民103.12] |
頁次 | 頁(45)1-(45)33 |
分類號 | 414.5 |
語文 | chi |
關鍵詞 | 蟾蜍靈; 發炎反應; 訊息傳遞; Bufalin; Inflammation; Signal transduction; |
中文摘要 | 不論在生物醫學及中國傳統醫學的領域中,疾病與疾病導致的基因改變以及藥物的作用等三面向的關連一直是很重要的研究目標。在實驗室的層級中如何發現適用的中草藥及如何釐清中藥作用的標的分子及相關機轉一直是很重要的工作。中藥的特色之一是常具有既能抗發炎又能增強免疫力的生物活性。而發炎反應是全身各組織均會發生的重要生理功能,然而失調的發炎反應卻也會對身體造成危害,例如敗血症即是臨床上常見且非常嚴重的發炎症狀。蟾酥,是蟾蜍毒腺的萃取物並作為主要在血癌細胞上的抗癌因子。然而對蟾酥在細胞發炎反應的藥效則未有任何相關報導。 研究目的:將探索蟾酥成分中的蟾蜍靈對細胞發炎反應的影響,闡述其中所參與作用方式和分子機轉,並探討蟾蜍靈之標的基因。 研究方法:採用細胞培養模式並以結合細胞學、生物化學及分子生物學的觀點去評估蟾蜍靈在細胞發炎上一連串反應中兩種非常重要的發炎分子,第二型環氧化酵素及細胞介白素第八之傳訊核醣核酸與蛋白質上的作用。我們也採取選擇性的有絲分裂活化蛋白質激酶及 NF-κB訊息路徑的抑制劑。生物晶片分析的運用也用於揭露蟾蜍靈可能作用的目標基因。 結果與討論:我們的實驗結果證明蟾蜍靈增強了蛋白質激酶 C所引發的第二型環氧化酵素及白細胞介素第八的傳訊核醣核酸與蛋白質表現。蟾蜍靈可能藉作用在有絲分裂活化蛋白質激酶及 NF-κB訊息路徑上來增強有絲分裂活化蛋白質激酶磷酸化及 NF-κB往細胞核之轉位,進而調控蛋白質激酶 C所誘發的細胞發炎。蟾蜍靈亦能調控許多除了發炎有關外之其他新穎的基因群。綜而觀之,我們的發現的確擴展了對蟾蜍靈是否能以及蟾蜍靈是如何貢獻在細胞層面上的發炎過程的作用。未來若在動物模式上進一步延續相關研究,勢必會更深入認知蟾蜍靈在整個生物體上的角色,並進而提供蟾蜍靈在人體健康上維護上的寶貴資訊及方法。 |
英文摘要 | Interaction between diseases, disease-modifying genes, and herbal modulators of those gene products has been a top aim of biomedical research as well as the core of Chinese medicine. It has been a critical goal in lab to locate herbal molecules with identified novel functions, to delineate the related mechanisms, and to identify the target molecules or genes of the pure herbal molecules. One feature of Chinese medicine is that it is able to perform an anti-inflammatory and/or to promote body immune activities. Inflammation is a normal physiological process, which occurs in a variety of tissues; however, unharmonious inflammation may result in devastating outcomes, such as sepsis. Bufadienolides are extracts of toxic glands from toads, and used as anticancer agents mainly on leukemia cells. However, the potential contribution of bufadienolides to the inflammatory process remains mostly unclear. AIM: This project is to investigate a pure component of bufadienolides, bufalin to evaluate its effects on cellular inflammatory cascades, to pinpoint the involving molecular mechanisms, and to identify its potential target genes in a global scope. METHODS: We used cell culture model to evaluate the impacts of bufalin on cellular inflammatory cascades from cellular, biochemical and molecular biological aspects by examining two critical inflammatory molecules cyclooxygenase-2 (COX-2) and interleukin 8 (IL-8) at mRNA and protein levels. We also adopted selective chemical inhibitors of MAPKs and NF-κB signal pathways. The microarray assay was also used to unveil the potential target genes of bufalin. RESULTS and DISSCUSSION: Our data demonstrated that bufalin enhanced PKC-induced CO-2 and IL-8 expression at protein and mRNA levels. Bufalin may target to MAPK as well as NF-κB pathways to regulate PKC-induced cellular inflammation by enhancing MAPK phosphorylation and nuclear NF-κB translocation to perform its roles in inflammation. Bufalin can regulate many more novels genes, other than inflammation-related ones. Our findings indeed expand our knowledge of if and how bufalin contributes to the inflammatory process in a cellular scope. Future work in animal model should further the whole picture of bufalin in a whole body, so as to provide valued information and options for bufalin to human health. |
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