Radicicol抑制內毒素誘發人類單核球細胞產生發炎細胞激素及基質金屬蛋白酶之機轉

在發炎疾病中，活化的人類單核球會製造、分泌許多的前發炎細胞激素(pro-inflammatory cytokines)及基質金屬蛋白酶(matrix metalloproteinases, MMPs)。本次研究欲探討從真菌Nectria balsamea #YMJ94052402萃取而來的天然物radicicol對人類單核球細胞釋放MMPs及前發炎細胞激素之影響與機轉。實驗結果顯示，radicicol 可以抑制腫瘤壞死因子???n?n( tumor necrosis factor-alpha, TNF-alpha)、白細胞介素1beta(interleukin-1beta, IL-1beta)或脂多醣體 (lipopolysaccharide, LPS)誘發人類單核球細胞製造及釋放MMP-9。利用西方墨點法，發現radicicol有濃度梯度效應的降低受LPS活化THP-1細胞的MMP-9蛋白質表現。Radicicol是一種熱休克蛋白90 (heat shock protein 90, HSP90) 抑制劑，發現其促進HSP70蛋白質的表現間接證明了抑制HSP90活性的能力。利用反轉錄-聚合酵素連鎖反應(Reverse transcription-polymerase chain reaction, RT-PCR) ，發現radicicol可以減少活化的THP-1之MMP-9的mRNA表現。在受到LPS刺激的THP-1細胞中，radicicol有意義反轉inhibitor-kappaB-alpha(IkappaB-alpha)的降解作用，並且發現radicicol能夠促進Akt降解。除此之外，利用酵素連結免疫吸附分析法(Enzyme-Linked Immunosorbent Assay, ELISA)發現 radicicol依濃度梯度效應降低受LPS活化之THP-1細胞釋放TNF-alpha或IL-6。綜合結論，本研究顯示radicicol可藉由調控NF-kappaB或PI3K/Akt pathway抑制MMP-9的表現及前發炎細胞激素的製造，並提供未來發展抗發炎藥物之新的方向。

關鍵字

內毒素；發炎細胞激素；基質金屬蛋白酶熱休克蛋白；單核球細胞

並列摘要

There is much evidence indicating that activated human monocytes / macrophages synthesize, and secrete several pro-inflammatory cytokines and matrix metalloproteinases (MMPs), and participate in the inflammatory diseases. In this study, we investigated the effects and mechanism of radicicol, as a natural product, extracted from Nectria balsamea #YMJ94052402, on human monocytic MMPs and pro-inflammatory cytokine expression. Radicicol exhibited concentration-dependent inhibition of MMP-9 enzymatic activation induced by tumor necrosis factor-alpha(TNF-alpha), interleukin-1beta(IL-1beta) or lipopolysaccharide (LPS) in human monocytic THP-1 cells. Western blot analysis showed that radicicol concentration-dependently suppressed monocytic MMP-9 protein expression. Radicicol is a heat shock protein 90 (HSP90) inhibitor. We found that radicicol enhanced HSP70 expression, and that indicated the inhibition ability of HSP90 in LPS-activated monocytic cells. Besides, reverse transcription- polymerase chain reaction (RT-PCR) showed that radicicol concentration-dependently suppressed monocytic MMP-9 mRNA level. Furthermore, Western blot analysis showed that radicicol could significantly reduce the degradation of inhibitor-kappaB-alpha (IkappaB-alpha) induced by LPS. In addition, we found that radicicol showed reduction of Akt expression. On the other hand, enzyme-linked immunosorbent assay (ELISA) showed that radicicol presented an inhibitory effect on TNF-alpha or IL-6 release in a concentration-dependent manner. In conclusion, we demonstrate that radicicol might involve interference with the NF-kappaB or PI3K/Akt pathway to attenuate MMP-9 expression and production of pro-inflammatory cytokines, and provide new opportunities for the development of new anti-inflammatory durgs.

並列關鍵字

radicicol ； Inflammatory Cytokine ； Matrix Metalloproteinase-9 ； heat shock protein ； monocyte

參考文獻

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Radicicol抑制內毒素誘發人類單核球細胞產生發炎細胞激素及基質金屬蛋白酶之機轉

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