液化澱粉芽孢桿菌胞外多醣對人類單核球細胞株THP-1之抗發炎效果與機制探討

The anti-inflammatory effect and mechanism of exopolysaccharides from Bacillus amyloliquefaciens on human monocyte cell line THP-1

林芸宇(Lin Yun-Yu)

指導教授 ： 鄭雪玲

國立屏東科技大學/農學院/生物科技系/碩士(2020年)

https://doi.org/10.6346/NPUST202100078

摘要

於前期研究中，液化澱粉芽孢桿菌（Bacillus amyloliqufaciens）amy-1的胞外多醣（exopolysaccharides, EPS）於動物實驗被發現具有抗發炎效果，但機制不明。於是本論文以人類單核球細胞株THP-1為模型，分析EPS對發炎中的巨噬細胞之抗發炎效果與分子機制。方法是以12-O-tetradecanoylphorbol (TPA)誘導THP-1細胞分化為M0期，再以lipopolysaccharides (LPS)誘導細胞分化為發炎狀態的M1期，分析LPS與EPS共處理時發炎的效果與機制。結果，EPS明顯抑制LPS所誘導的iNOS, TNF-α, IL-6等發炎因子的表現。EPS也抑制LPS所誘導的IκB kinase (IKK) / nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB)路徑的活化。同時EPS也抑制mitogen-activated protein kinase (MAPK)中ERK的活化，但較高濃度的EPS卻活化JNK及p38。另外，EPS能活化抗氧化酵素heme oxygenase-1 (HO-1)的表現，並抑制ROS的生成。研究更進一步的發現，EPS抗氧化的效果至少與活化p38有關。結論是，EPS能抑制LPS所引起的發炎反應，機制與IKK/ NF-κB路徑的抑制有關。同時EPS能藉由活化p38來活化保護性的抗氧化機制，以降低發炎對細胞的傷害。

關鍵字

抗發炎 ； Bacillus amyloliqufaciens ； exopolysaccharides ； THP-1 ； NF-κB ； MAPK

並列摘要

The exopolysaccharides (EPS) of Bacillus amyloliqufaciens amy-1 were found to have anti-inflammatory effects in animal tests, but the underlying mechanism is unknown. Therefore, this study used the human monocyte cell line THP-1 as a model to investigate the anti-inflammatory effect and molecular mechanism of EPS on macrophages. The cells were induced to differentiate into M0 phase by 12-O-tetradecanoylphorbol (TPA). Lipopolysaccharides (LPS) was then used to induce the differentiation of THP-1 cells into the pro-inflammatory M1 phase. This model was used to analyze the effect of EPS. Consequently, EPS significantly inhibited LPS-induced expression of iNOS, TNF-α and IL-6. Meanwhile, EPS inhibited the activation of IκB kinase (IKK) /nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway by LPS. Moreover, EPS also suppressed the activation of mitogen-activated protein kinase (MAPK), ERK, but higher concentrations of EPS activated JNK and p38. In addition, EPS activated the expression of the antioxidant enzyme heme oxygenase-1 (HO-1) and reduced the generation of ROS. Furthermore, it was found that the antioxidation effect of EPS is at least correlated with the activation of p38. In conclusion, EPS inhibits LPS-induced inflammation, and the underlying mechanism is associated with the suppression of the IKK/NF-κB pathway. Meanwhile, EPS activate the protective antioxidant pathway by activating p38 to reduce cellular damages caused by inflammation.

並列關鍵字

Bacillus amyloliqufaciens ； exopolysaccharides ； THP-1 ； NF-κB ； MAPK

參考文獻

黃聖達，分析益生菌胞外多醣的降血糖及抗發炎效果，國立屏東科技大學生物科技系碩士論文，2015。

Arulselvan, P., Fard, M.T., Tan, W.S., Gothai, S., Fakurazi, S., Norhaizan, M.E., and Kumar, S.S. (2016). Role of antioxidants and natural products in inflammation. Oxidative Medicine and Cellular Longevity 2016.

Ayroldi, E., Cannarile, L., Migliorati, G., Nocentini, G., Delfino, D.V., and Riccardi, C. (2012). Mechanisms of the anti‐inflammatory effects of glucocorticoids: genomic and nongenomic interference with MAPK signaling pathways. The FASEB Journal 26, 4805-4820.

Baharav, E., Mor, F., Halpern, M., and Weinberger, A. (2004). Lactobacillus GG bacteria ameliorate arthritis in Lewis rats. The Journal of Nutrition 134, 1964-1969.

Barendsen, N., Mueller, M., and Chen, B. (1990). Inhibition of TPA-induced monocytic differentiation in THP-1 human monocytic leukemic cells by staurosporine, a potent protein kinase C inhibitor. Leukemia Research 14, 467-474.