查耳酮衍生物AN07經由活化PPAR-γ減緩脂多醣在小鼠巨噬細胞株RAW264.7引起的發炎及氧化壓力

A novel chalcone derivative AN07 attenuates lipopolysaccharide-induced inflammation and oxidative stress via activating PPAR-γ in RAW264.7 macrophages

廖智堯(Zhi-Yao Liao)

指導教授 ： 羅怡卿

高雄醫學大學/醫學院/醫學系藥理學科碩士班/碩士(2016年)

摘要

查耳酮(chalcone)是一種植物類黃酮(flavonoids)，具有多種藥理活性，包括抗發炎和抗氧化作用。因過氧化物酶體增生活化接受器(peroxisome proliferator-activated receptor, PPAR) 在發炎症狀調節過程中扮演關鍵角色，而在之前的研究中發現，查耳酮衍生物AN07可透過活化PPAR-γ而具有治療動脈粥狀硬化的潛力。因此，本研究，我們將探討AN07是否能減緩脂多醣(lipopolysaccharide, LPS)在小鼠巨噬細胞株RAW264.7所產生的發炎反應及氧化壓力，並觀察PPAR-γ 是否參與ΑΝ07的保護作用。抗發炎作用的研究結果顯示，AN07能有效抑制脂多醣活化小鼠巨噬細胞株RAW264.7之誘導型一氧化氮合酶(iNOS)、環氧合酶-2 (COX-2)及降低抑制蛋白IκBα 磷酸化。同時，AN07也能有效減少脂多醣誘導的一氧化氮(NO)產生。而AN07抗氧化作用的研究結果顯示，AN07不僅抑制脂多醣對醣蛋白(glycoprotein 91, gp91phox)的誘導表現，它也能抑制活性氧化物質(reactive oxygen species, ROS)的產生，活化NF-E2相關因子2 (nuclear factor erythroid-2-related factor, Nrf2)和第一型血色素氧化酵素(heme oxygenase 1, HO-1)表現，以及增加穀胱甘肽 (glutathione, GSH)生成。為了證明PPAR-γ是否參與ΑΝ07的抗發炎及抗氧化作用，本研究使用PPAR-γ 拮抗劑GW9662檢視AN07產生的COX-2抑制及Nrf2的活化作用。實驗結果顯示，GW9662明顯減少AN07抑制脂多醣對COX-2的誘導表現及AN07活化Nrf2的作用。綜合以上研究結果，AN07能減緩脂多醣在小鼠巨噬細胞株RAW264.7所產生的發炎反應及氧化壓力，而其抗發炎及抗氧化作用至少有部分是藉由活化PPAR-γ 所產生。因此，AN07極具潛力成為新的抗發炎及抗氧化藥物。

關鍵字

查耳酮衍生物 ； 脂多醣 ； 發炎 ； 氧化壓力

並列摘要

Chalcone is a class of plant flavonoids with multiple pharmacological activities, including anti-inflammatory and anti-oxidative effects. The peroxisome proliferator-activated receptor (PPAR) regulatory pathway plays a critical role in the regulation of inflammation. A chalcone derivative AN07 was reported its novel therapeutic potential against atherosclerosis through PPAR- γ activation. In this study, we investigated the effects of AN07 on lipopolysaccharide (LPS)-induced inflammation and oxidative stress, and the role of PPAR- γ in AN07 protection in macrophage RAW264.7. Results indicated that AN07 inhibited LPS-induced expressions of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and phosphor - nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor alpha (p-IκBα) in RAW264.7. LPS-induced nitric oxide (NO) production was also attenuated by AN07. Moreover, AN07 not only attenuated gp91phox expression and ROS production, but it also enhanced expression of Nrf2, HO-1 activation and production of glutathione (GSH) in LPS-stimulated RAW264.7 cells. Furthermore, PPAR-γ antagonist GW9662 significantly attenuated AN07-induced COX-2 inhibition and Nrf2 activation in LPS-stimulated RAW264.7 cells. Taken together, the present results demonstrate that AN07 exerts anti-inflammatory and anti-oxidant effects in macrophages, at least in part, through partial PPAR- γ activation, suggesting AN07 possesses a potential to be an anti-inflammatory and anti-oxidant agent.

並列關鍵字

chalcone derivative ； lipopolysaccharide ； inflammation ； oxidative stress

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