染布青萃出物透過血基質氧化酶-1抑制脂多醣誘發的巨噬細胞發炎反應

Indigofera suffruticosa Mill extracts Suppression of Lipopolysaccharide-Induced Inflammation of Macrophages through the Heme Oxygenase-1

梁純慈(Chun-Tzu Liang)

指導教授 ： 劉德中

中山醫學大學/健康管理學院/營養學研究所/碩士(2011年)

https://doi.org/10.6834/CSMU.2011.00151

摘要

染布青(Indigofera suffruticosa Mill)，別名大菁、大青、番菁、野青樹、假藍靛、菁子、青仔、木藍、野木藍等，染布青的莖葉含靛(indirubin)可提煉藍色染料，大多野生於台灣山區。有研究指出染布青具有抗真菌及抗細菌活性之功能，並可有效的抑制老鼠腫瘤之生長。本研究使用小鼠巨噬細胞(RAW264.7)在脂多醣(Lipopolysaccharide, LPS)誘發發炎反應的模式下，同時處理染布青水溶液或乙醇萃出物，結果顯示染布青萃出物對細胞存活率並無顯著之影響(p>0.05)。由一氧化氮含量、一氧化氮合成酶蛋白質和mRNA表現之分析發現染布青萃出物可有效抑制LPS誘發之發炎反應(p<0.05)，隨著萃出物濃度的提升，抑制之作用有逐漸增強之現象。另外，處理染布青萃出物最高劑量，亦可有效抑制促發炎激素TNF-α生成。進一步探討染布青萃出物對於體內抗氧化機制之血基質氧化酶(Heme oxgenase,HO)-1的表現，結果顯示染布青萃出物可有效提升HO-1蛋白質和mRNA表現量(p<0.05)。在預處理染布青萃出物之實驗模式中，當HO-1提升到最大表現量時，再給予LPS刺激，染布青萃出物可有效抑制LPS誘發之IL-1β蛋白質表現(p<0.05)，隨著萃出物濃度的提升抑制表現的效果逐漸增強。由實驗結果證實：染布青水溶液或乙醇萃出物可能藉由增加HO-1表現，進而抑制LPS所誘發的發炎反應。

關鍵字

染布青 ； 脂多醣 ； 小鼠巨噬細胞RAW264.7 ； 一氧化氮 ； 血基質氧化酶

並列摘要

Indigofera suffruticosa Mill (I. suffruticosa) contains indigo in stems and leaves. Previous studies showed that I. suffruticosa has anti-fungal and anti-bacterial activities, and can effectively inhibit the growth of mouse tumors. In the present study, we used mouse RAW264.7 macrophage cell which treated with Lipopolysaccharide (LPS) to induce inflammation, the treatment of aqueous extracts (WE) or ethanol extracts (EE) of I. suffruticosa showed no significant effect (p> 0.05) in cell survival. In the analysis of nitric oxide (NO) production, the expression of nitric oxide synthase protein and mRNA, we found that extracts of I. suffruticosa can be effectively inhibited LPS-induced inflammation (p <0.05) with a concentration dependent manner. In addition, treatment of I. suffruticosa extracts can also inhibit the production of pro-inflammatory cytokine TNF-α. Further studies showed the extracts of I. suffruticosa can also enhance HO-1 protein and mRNA expression (p <0.05). In the pre-treatment of extracts, when the HO-1 expression raised to the maximum amount, extracts of I. suffruticosa can effectively inhibit LPS-induced IL-1β protein expression (p < 0.05) with a dose-dependent manner. These results confirmed that aqueous extracts (WE) or ethanol extracts (EE) of I. suffruticosa may be inhibit LPS-induced inflammation through the expression of HO-1.

並列關鍵字

Indigofera suffruticosa Mill ； Lipopolysaccharide ； mouse RAW264.7 macrophage cell ； nitric oxide ； Heme Oxygenase

參考文獻

de Sousa Falcao, H., Leite, J.A., Barbosa-Filho, J.M., de Athayde-Filho, P.F., de Oliveira Chaves, M.C., Moura, M.D., Ferreira, A.L., de Almeida, A.B., Souza-Brito, A.R., de Fatima Formiga Melo Diniz, M., et al. (2008). Gastric and duodenal antiulcer activity of alkaloids: a review. Molecules 13, 3198-3223.

Vital, A.L., Goncalo, M., Cruz, M.T., Figueiredo, A., Duarte, C.B., and Lopes, M.C. (2003). Dexamethasone prevents granulocyte-macrophage colony-stimulating factor-induced nuclear factor-kappaB activation, inducible nitric oxide synthase expression and nitric oxide production in a skin dendritic cell line. Mediators Inflamm 12, 71-78.

何玉鈴, 郭昭麟, 高國清, 陳忠川, and 張永勳 (2001). 板藍根藥材之鑑別研究. J Chin Med 12, 267-283.

趙文婉, 郭悅雄, and 林璧鳳 (2007). 以啟動子轉殖細胞株篩檢調節免疫力之中藥的模式建立. Taiwanese Journal of Agricultural Chemistry and Food Science 45, 193-205.

Chen, C.Y., Jang, J.H., Li, M.H., and Surh, Y.J. (2005). Resveratrol upregulates heme oxygenase-1 expression via activation of NF-E2-related factor 2 in PC12 cells. Biochem Biophys Res Commun 331, 993-1000.