Effect and mechanism of IKK-2 receptor blocker on corneal stromal fibroblasts

doi:10.3969/j.issn.2095-4344.2015.29.019

Abstract

Abstract:

BACKGROUND: Corneal stromal fibroblasts have been shown to express interleukin-8 in the stimulation of lipopolysaccharide. Different reactions of fibroblasts to lipoprotein or other inflammatory mediators may constitute different characteristics of different tissues in the inflammatory response.
OBJECTIVE: To observe the inhibitory effect of IKK-2 receptor blocker TPCA-1 on human corneal stromal fibroblasts to secrete inflammatory cytokines, chemokines and cell adhesion molecules under the stimulation of lipopolysaccharide and its signal transduction pathway, to compare with dexamethasone, and to explore alternative or synergistic effects after their combination.
METHODS: This study measured the secretion of interleukin-1, tumor necrosis factor alpha, interleukin-6, and interleukin-8 from cultured human corneal stromal fibroblasts under the action of basic state and lipopolysaccharide, and their changes after the intervention with IKK-2 receptor blocker TPCA-1 and dexamethasone. We also detected expression level of intercellular adhesion molecule-1 and interleukin-6 in cell surface, and verified the changes in expression levels of intercellular adhesion molecule-1, interleukin-6, and interleukin-8 from mRNA level, as well as examined the expression of nuclear factor kappa B under above conditions.
RESULTS AND CONCLUSION: Lipopolysaccharide induced the secretion of interleukin-6, interleukin-8 and intercellular adhesion molecule-1 from human corneal stromal fibroblasts, and their expressions were increased. IKK-2 receptor blocker TPCA-1 could inhibit above effects. The inhibitory effect of TPCA-1 was done by blocking nuclear transcription factor kappa B signal transduction, and was identical to the inhibitory effect of dexamethasone, but by different signal transduction pathway. These data indicate that IKK-2 receptor blocker and dexamethasone have synergistic effects. IKK-2 receptor blocker may become an alternative to dexamethasone in treatment of infectious corneal diseases.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Key words: lipopolysaccharide, Fibroblasts, Dexamethasone

CLC Number:

R318

Zhong Wei, Jiang Hong, Zhao Jing. Effect and mechanism of IKK-2 receptor blocker on corneal stromal fibroblasts [J]. Chinese Journal of Tissue Engineering Research, 2015, 19(29): 4688-4694.

Figures/Tables 4

References

[1] 宋秀君.重视角膜基质细胞的作用[J].中华实验眼科杂志, 2012, 29(12):1057-1060．

[2] 陆燕,孟虎,侯培莉,等.来源于全飞秒激光小切口微透镜摘除术的基质层人角膜成纤维细胞的原代培养与鉴定[J].眼科新进展，2013,33(9):801-803.

[3] Brouty-Boye’ D, Pottin-Cle’ menceau C,et al. Chemokines and CD40 expression in human fibroblasts.Eur J Immunol. 2000;30:914-919.

[4] Fukuda K, Fujitsu Y, Seki K,et al.Differential expression of thymus- and activation-regulated chemokine (CCL17) and macrophage-derived chemokine (CCL22) by human fibroblasts from cornea, skin, and lung.J Allergy Clin Immunol. 2003;111:520-526.

[5] Fukuda K,Ishida W1,Tanaka H2,Harada Y1,Fukushima A1.Inhibition by rebamipide of cytokine-induced or lipopolysaccharide-induced chemokine synthesis in human corneal fibroblasts.Br J Ophthalmol.2014;98(12):1751-1755.

[6] Koyama S, Sato E, Nomura H, et al. The potential of various lipopolysaccharides to release monocyte chemotactic activity from lung epithelial cells and fibroblasts. Eur Respir J.1999; 14:545-552.

[7] Wilson SE,Mohan RR,Mohan RR,et al.The corneal wound healing response: cytokine-mediated interaction of the epithelium, stroma, and inflammatory cells. Prog Retinal Eye Res.2001;20:625-637.

[8] Birrell MA, Hardaker E, Wong S, et al.I-B kinase-2 inhibitor blocks inflammation in human airway smooth muscle and a rat model of asthma. Am J Respir Crit Care Med. 2005;172: 962-971.

[9] Gaddipati S, Lu Q, Kasetti RB, et al.IKK2 Inhibition Using TPCA-1-Loaded PLGA Microparticles Attenuates Laser-Induced Choroidal Neovascularization and Macrophage Recruitment.PLoS One. 2015;10(3):e0121185.

[10] Stinson LF,Ireland DJ,Kemp MW,et al.Effects of cytokine-suppressive anti-inflammatory drugs on inflammatory activation in ex vivo human and ovine fetal membranes. Reproduction. 2014;147(3):313-320.

[11] Saika S,Miyamoto T,Yamanaka O,et al.Therapeutic effect of topical administration of SN50,an inhibitor of nuclear factor-B,in treatment of corneal alkali burns in mice. Am J Pathol. 2005;166:1393-1403.

[12] Saika S, Ikeda K, Yamanaka O, et al. Expression of Smad7 in mouse eyes accelerates healing of corneal tissue after exposure to alkali.Am J Pathol. 2005;166:1405-1418.

[13] Li Q, Fukuda K, Lu Y, et al. Enhancement by neutrophils of collagen degradation by corneal fibroblasts. J Leukoc Biol. 2003;74:412-419.

[14] Lu Y, Liu Y, Fukuda K, et al.Inhibition by Triptolide of Chemokine, Proinflammatory Cytokine, and Adhesion Molecule Expression Induced byLipopolysaccharide in Corneal Fibroblasts. Invest Ophthalmol Vis Sci.2006;47 (9): 3796-880.

[15] De Bosscher K,Vanden Berghe W,et al.The interplay between the glucocorticoid receptor and nuclear factor-Boractivator protein-1: molecular mechanisms for gene repression.Endocr Rev.2003;24:488-522.

[16] Almodóvar-García K, Kwon M, Samaras SE,et al. ANKRD1 acts as a transcriptional repressor of MMP13 via the AP-1 site. Mol Cell Biol. 2014;34(8):1500-11.

[17] Shaulian E, Karin M. AP-1 in cell proliferation and survival. Oncogene. 2001,20: 2390-2400.

[18] Gonza′lez MV, Jime‘nez B, Berciano MT, et al. Glucocorticoids antagonize AP-1 by inhibiting the activation/phosphorylation of JNKwithout affecting its subcellular distribution.J Cell Biol.2000;150:1199-1208.

[19] Chakraborti S, Mandal M, Das S,et al. Regulation of matrix metalloproteinases: an overview. Mol Cell Biochem.2003; 253:269-285.

[20] Orita T, Kimura K, Nishida T,et al. Cytokine and chemokine secretion induced by poly(I:C) through NF-κB and phosphoinositide 3-kinase signaling pathways in human corneal fibroblasts. Curr Eye Res. 2013 Jan;38(1):53-9.

[21] Li W, Zheng S, Tang C,et al. JNK-AP-1pathway involved in interleukin- 1-induced calcitonin gene-related peptide secretion in human type II alveolar epithelial cells. Peptides. 2007;28:1252-1259.

[22] Liacini A,Sylvester J,Li WQ,et al.Inhibition of interleukin-1-stimulated MAP kinases, activating protein-1 (AP-1) and nuclear factor kappaB (NF-B) transcription factors down-regulate smatrix metalloproteinase gene expression in articular chondrocytes. Matrix Biol.2002;21:251-262.

[23] 赵伟,周瑾,郭梦翔,等.地塞米松对碱烧伤后大鼠角膜核转录因子-κB活化的影响[J].眼科新进展,2013,33(7):614-617.

[24] Takeda K, Takeuchi O, Tsujimura T, et al. Limb and skin abnormalities in mice lacking IKK. Science.1999;284: 313-316.