共軛亞麻油酸對TNF-alpha誘發小鼠淋巴結內皮細胞SVEC4-10表現黏著分子MAdCAM-1之影響

發炎性腸道疾病主要是指克隆氏症及潰瘍性結腸炎。當罹患發炎性腸道疾病時，腸道內皮細胞表面會表現MAdCAM-1，使淋巴球附著至病灶，加重病情。本實驗利用TNF-alpha刺激SVEC4-10小鼠內皮細胞，模仿罹患發炎性腸道疾病時，共軛亞麻油酸影響MAdCAM-1生合成的效應及其相關機轉。MTT 分析細胞存活率的結果顯示，以20 ng/ml TNF-alpha及100 microM CLA單獨處理或分別處理24小時，皆不減少SVEC4-10小鼠內皮細胞的存活率，若CLA濃度大於100 microM則會使細胞存活率下降，故本實驗以100 microM CLA為實驗用量。20 ng/ml TNF-alpha可誘發MAdCAM-1蛋白質表現，而100 microM CLA則可顯著抑制TNF-alpha誘發的MAdCAM-1蛋白質表現。進一步實驗分析結果也顯示在20 ng/ml TNF-alpha的刺激下，100 microM CLA可以顯著抑制PKB及JNK的磷酸化、IkappaB-alpha的降解及NF-kappaB與kappaB site的結合。綜合以上結果可知，CLA可能是透過PKB及JNK的訊息傳遞路徑，抑制轉錄因子NF-kappaB進而影響MAdCAM-1蛋白質生合成。

關鍵字

SVEC4-10小鼠內皮細胞；共軛亞麻油酸； MAdCAM-1 ； PKB ； JNK ； NF-kappaB

並列摘要

Inflammatory bowel disease (IBD) commonly refers to ulcerative colitis (UC) and Crohn’s disease (CD), which are chronic inflammatory diseases of the GI tract. The endothelial cells of intestine may secrete mucosal adressin cell adhesion molecule-1 (MAdCAM-1), and attract the local lymphocytes to the injury tissue in the patient’s of IBD. In this experiment, we use TNF-alpha-induced SVEC4-10 cells to produce mimic IBD, the objective of this experiment is to explore the influence of conjugated linoleic acid on MAdCAM-1. MTT assay showed that neither addition of 20 ng/ml TNF-alpha nor 100 microM CLA treatment express in SVEC4-10 cells did not influence the viability. 100 microM CLA significantly inhibited TNF-alpha-induced MAdCAM-1 expression (p<0.05), as well as phosphorylation of PKB compared with TNF-alpha treatment only exogenous addition of SVEC4-10 cells (p<0.05). Further experience has demonstrated that 100 microM CLA treatment inhibited TNF-alpha-induced IkappaB-alpha degradation and NF-kappaB nucleus protein-DNA binding activity. This experience also showed that CLA could significantly inhibit phosphorylation of c-Jun N-terminal kinase (JNK) (p<0.05). Taken together, these results suggest that CLA inhibits TNF-alpha-induced MAdCAM-1 expression in SVEC4-10 cells is resulted from inhibition of NF-kappaB activation. Moreover, this result indicated CLA is related with inhibition of JNK phosphorylation. Moreover, our result also indicated that CLA regulated TNF-alpha induce NF-kappaB activation is through PKB, TNF-alpha related pathway.

並列關鍵字

SVEC4-10 cells ； conjugated linoleic acid ； MAdCAM-1 ； PKB ； JNK ； NF-kappaB

參考文獻

Alexander, J. S., and Ando, T. (2005). Density-dependent control of MAdCAM-1 and chronic inflammation. Focus on "Mechanisms of MAdCAM-1 gene expression in human intestinal microvascular endothelial cells". Am. J. Physiol. Cell Physiol. 288, C243-244.

Aranda, A., and Pascual, A. (2001). Nuclear hormone receptors and gene expression. Physiol. Rev. 81, 1269-1304.

Arnott, I. D., Watts, D., and Satsangi, J. (2003). Azathioprine and anti-TNF alpha therapies in Crohn's disease: a review of pharmacology, clinical efficacy and safety. Pharmacol. Res. 47, 1-10.

Baldwin, A. S., Jr. (2001). Series introduction: the transcription factor NF-kappaB and human disease. J. Clin. Invest. 107, 3-6.

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共軛亞麻油酸對TNF-alpha誘發小鼠淋巴結內皮細胞SVEC4-10表現黏著分子MAdCAM-1之影響

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