- 學位論文
有機合成物與球薑酮之體外抗發炎作用及其分子機轉研究
Effects of synthetic organic compounds and zerumbone on in vitro anti-inflammatory actions and their molecular mechanisms
摘要
發炎反應為生物體對抗外來與內生性刺激之防禦機制。長期處於慢性發炎狀態下卻可能誘發癌症、心血管疾病和第二型糖尿病等多種慢性疾病之發生。肥胖即為一種慢性發炎疾病，並刺激巨噬細胞浸潤於脂肪組織中，藉由交互作用釋出大量的發炎因子，例如：腫瘤壞死因子 (tumor necrosis factor-α, TNF-α)、介白素-6 (interleukin-6, IL-6) 及單核球趨化蛋白質-1 (monocyte chemoatrractant protein-1, MCP-1)，進而增加第二型糖尿病和動脈粥狀硬化之罹患風險。在本研究中所使用之樣品，有機合成物與球薑酮 (zerumbone)，過去研究皆指出兩者在抗癌、抗氧化與抗發炎等生理活性上具良好之功用。但目前利用有機合成物與zerumbone探討發炎與肥胖發炎之相關研究仍是缺乏。因此，本研究內容分為兩部分：(一) 探討有機合成物在脂多醣 (lipopolysaccharide, LPS) 誘導RAW 264.7巨噬細胞發炎與TNF-α誘導3T3-L1脂肪細胞發炎之影響及其分子機轉探討 (二) 探討zerumbone在LPS誘導RAW 264.7巨噬細胞發炎與TNF-α誘導3T3-L1脂肪細胞發炎之影響及其分子機轉探討。第一部份結果顯示，在24種有機合成物中，以BZT-21和BZT-26具有較佳抑制LPS誘導RAW 264.7巨噬細胞分泌一氧化氮之作用。在細胞激素結果顯示，有機合成物BZT-21和BZT-26皆可顯著降低PGE2、IL-6和MCP-1之分泌。在基因表現上，有機合成物BZT-21和BZT-26可顯著降低發炎相關基因 (TLR-4、p65、JAK-2、STAT-3、IL-6、IL-1 bata、MCP-1、COX-2和iNOS) 之表現。此外，以BZT-21和BZT-26處理TNF-α誘導3T3-L1脂肪細胞發炎反應，其可顯著降低IL-6和MCP-1 之發炎細胞激素分泌，並抑制發炎相關指標 (IL-6、MCP-1、CRP和NF-kappa B) 之基因表現，並提升AMPK、SIRT-1與Adiponectin之基因表現。第二部份結果顯示，以zerumbone處理LPS誘導RAW 264.7巨噬細胞發炎反應，其可顯著降低一氧化氮、PGE2、IL-6、IL-1 bata、MCP-1和TNF-α之分泌。在基因表現上，zerumbone可顯著降低發炎相關 (TLR-4、p65、JAK-2、STAT-3、IL-6、IL-1 beta、MCP-1、TNF-α、COX-2和iNOS) 之基因表現。以zerumbone處理TNF-α誘導3T3-L1脂肪細胞發炎，其可顯著降低IL-6和MCP-1之發炎細胞激素分泌，並抑制發炎相關指標 (IL-6、MCP-1、CRP和NF-kappa B) 之基因表現，提升AMPK、SIRT-1與Adiponectin之基因表現。從上述結果顯示，有機合成物BZT-21、BZT-26與zerumbone皆可對LPS所誘導巨噬細胞與TNF-α誘導脂肪細胞兩者達到抑制發炎的反應。因此，有機合成物BZT-21、BZT-26與zerumbone皆具有良好抗發炎與改善肥胖發炎之作用。
並列摘要
The inflammatory response provides the first line of host defense against a variety of exogenous and endogenous stimuli. The progression of chronic inflammation to diseases (such as cancer, cardiovascular diseases, and metabolic diseases) is a complex process involving multiple biological factors. Obesity-associated chronic inflammation is characterized by macrophage infiltration while occurs mainly in adipose tissues, and leads to an increased release of pro-inflammatory cytokines, such as tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and macrophage chemo-attractant protein-1 (MCP-1). The overproduction of pro-inflammatory cytokines seem to be important causes of morbidity and some relative diseases, such as type 2 diabetes and atherosclerosis. Synthetic organic compounds and zerumbone have been reported to possess multiple biological functions, including anti-cancer, anti-oxidant, and anti-inflammatory activities. However, the anti-inflammatory effects of synthetic organic compounds and zerumbone in cell culture model remain unclear. Therefore, there are two topics included in this study, (1) Effects of synthetic organic compounds on LPS-induced inflammatory response in RAW 264.7 macrophages, and TNF-α-induced inflammatory response in 3T3-L1 adipocytes and their molecular mechanisms. (2) Effect of zerumbone on LPS-induced inflammatory response in RAW 264.7 macrophages, and TNF-α-induced inflammatory response in 3T3-L1 adipocytes and their molecular mechanisms. (1) The result showed that BZT-21 and BZT-26 had the highest inhibition on LPS-induced nitric oxide (NO) in RAW 264.7 macrophages among 24 synthetic organic compounds tested. Moreover, BZT-21 and BZT-26 inhibited LPS-induced inflammatory cytokines (IL-6, IL-1β, PGE2, and MCP-1) and inflammatory-related gene expressions (TLR-4, p65, JAK-2, STAT-3, IL-6, IL-1β, MCP-1, COX-2, and iNOS) in RAW 264.7 macrophages among the compounds tested. BZT-21 and BZT-26 also inhibited TNF-α-induced inflammatory cytokines (IL-6 and MCP-1), and inflammatory-related gene expressions (IL-6, MCP-1, CRP, and NF-κB), as well as increased the gene expressions of SIRT-1, AMPK, and Adiponectin in 3T3-L1 adipocytes. (2) The result showed that zerumbone inhibits LPS-induced inflammatory cytokines (NO, IL-6, IL-1β, PGE2, TNF-α, and MCP-1) and inflammatory-related gene expressions (TLR-4, p65, JAK-2, STAT-3, IL-6, IL-1β, MCP-1, TNF-α, COX-2, and iNOS) in RAW 264.7 macrophages. Moreover, zerumbone inhibited TNF-α-induced inflammatory cytokines (IL-6 and MCP-1), and inflammatory-related gene expressions (IL-6, MCP-1, CRP, and NF-κB), as well as increased the gene expressions of SIRT-1, AMPK, and Adiponectin in 3T3-L1 adipocytes. These results indicated that BZT-21, BZT-26, and zerumbone are suppressed LPS-induced inflammatory response in RAW 264.7 macrophages, and TNF-α-induced inflammatory response in 3T3-L1 adipocytes. Therefore, BZT-21, BZT-26, and zerumbone have great anti-inflammatory actions and improvement of obesity-related inflammatory response.