數株乳酸桿菌胞外多醣之抗發炎作用及抑制結腸癌細胞增生之研究

Effects of Exopolysaccharides Produced by Several Lactobacillus spp. on the Anti-inflammatory Activity and Antiproliferation of Colon Cancer Cells

李逸華(Yi-Hua Lee)

指導教授 ： 游若篍

國立臺灣大學/生物資源暨農學院/食品科技研究所/碩士(2011年)

https://doi.org/10.6342/NTU.2011.10806

摘要

發炎性腸道疾病 (Inflammatory bowel disease, IBD) 為一種慢性發炎疾病，長期罹患 IBD 之患者其罹患結腸癌之風險會增加，而益生菌和益生質對於 IBD 治療很有發展潛力。乳酸菌為腸道中之益生菌，對宿主健康有很大益處，而其部分原因可能與其所產之胞外多醣 (Exopolysaccharides, EPS) 的生理活性有關。本研究以具有對雙叉桿菌生長助生性之乳酸桿菌 (Lactobacillus acidophilus BCRC 14079、L. casei 01、L. delbrueckii subsp. bulgaricus BCRC 10696、L. plantarum BCRC 11697 及 L. rhamnosus GG BCRC 16000) 胞外多醣為材料，並利用脂多醣 (Lipopolysaccharide, LPS) 誘導小鼠巨噬細胞株 (RAW 264.7 cell line) 之三種發炎模式 (預防發炎模式、修復發炎模式及 EPS 與 LPS 共培養發炎模式) 進行，探討乳酸桿菌之胞外多醣其抗發炎作用。實驗結果顯示，乳酸桿菌胞外多醣在 EPS與 LPS 共培養發炎模式下，具有較佳之抗發炎效果，而五種乳酸桿菌胞外多醣皆具有清除 NO (Nitric oxide) 之能力，亦可抑制 NO 生成及 TNF-α 產生，且其效果具有濃度效應 (Dose-dependent)，其中以 L. plantarum BCRC 11697 產生之胞外多醣其抑制 NO 生成的效果最佳，故乳酸桿菌胞外多醣具有抗發炎之作用，有發展減緩因發炎而造成疾病之潛力。在抑制結腸癌細胞增生實驗中，五種乳酸桿菌胞外多醣作用 HT-29 細胞 48 小時皆有抑制增生之作用；另一方面，除了 L. delbrueckii subsp. bulgaricus BCRC 10696 的胞外多醣之外，其餘四種乳酸桿菌胞外多醣皆有抑制 Caco-2 細胞增生之作用，且隨著作用時間增加，其抑制增生效果越好。因此，乳酸桿菌之胞外多醣不但具有抗發炎作用亦可抑制結腸癌細胞增生之作用，具有發展治療 IBD 之潛力。

關鍵字

發炎性腸道疾病 (IBD) ； 胞外多醣 ； 抗發炎 ； RAW 264.7

並列摘要

Inflammatory bowel disease (IBD) was a chronic inflammatory disease, and long-time suffered from IBD could increase risk of colon cancer. Several lactic acid bacteria (LAB) were probiotics, the beneficical effects on human health of LAB could be attributable to positive physiological effect of exopolysaccharide (EPS). Study had shown that probiotics and prebiotics were showing increasing promise as treatments for IBD. We employed EPS produced by five Lactobacillus spp. (Lactobacillus acidophilus BCRC 14079、L. casei 01、L. delbrueckii subsp. bulgaricus BCRC 10696、L. plantarum BCRC 11697 and L. rhamnosus GG BCRC 16000), which had positive prebiotic effect on the growth of Bifidobacterium, as materials and three lipopolysaccharide (LPS)-induce inflammation models (preventive inflammation model, repairing inflammation model, and co-treated inflammation model) were established with murine macrophages (RAW 264.7 cell line) to evaluate the anti-inflammatory activity of EPS. Results revealed that all of EPS produced by Lactobacillus spp. had better anti-inflammatory activity in EPS and LPS co-treated inflammation model. All of EPS had NO scavenging activity, and could inhibit the production of TNF-α and nitric oxide (NO) in a dose-dependent manner. Especially, EPS of L. plantarum BCRC 11697 have the lowest IC50 (188.4 ± 3.0 μg/ml), shown that had the best inhibition of NO production. So, all of EPS had anti-inflammatory activity. Furthermore, all of EPS had anti-proliferation on HT-29 cell after 48 h incubation. Also, besides the EPS from L. delbrueckii subsp. bulgaricus BCRC 10696, four of EPS of Lactobasillus spp. could inhibit the proliferation of Caco-2 cells in a time-dependent manner. Conclusively, all of EPS produced by Lactobacillus spp. had anti-inflammatory activity and anti-proliferative activity of colon cancer cell line, so EPS had potential to develop treatment of IBD.

並列關鍵字

inflammatory bowel disease (IBD) ； exopolysaccharides (EPS) ； anti-inflammatory activity ； RAW 264.7 cell line

參考文獻

王姿以。2009。攝食菊醣改善人體腸胃道菌相並降低糞便萃取液對HT-29及Caco-2毒性之研究。國立臺灣大學食品科技所碩士學位論文。台北。台灣。

行政院衛生署。2011。99年國人主要死因統計。行政院衛生署。台北。台灣。

陳倩琪、彭宣融、廖麗玲。2007a。基因體研究對乳酸菌機能保健品之產業發展潛力。農業生技產業季刊。11：52-29。

陳慶源、黃崇真、邱雪惠、廖啟成。2007b。乳酸菌之保健功效與產品開發。農業生技產業季刊。11：60-68。

陳穗葶。2005。數株乳酸菌胞外多醣對雙叉桿菌生長之助生性研究。國立臺灣大學食品科技研究所碩士學位論文。台北。台灣。