Abstract
This study focused on the effects of aspirin on lipopolysaccharide (LPS)-induced expression of phosphoinositide 3 kinase (PI3K)/protein kinase B (Akt), extracellular signal-regulated protein kinase (ERK), nuclear factor-κB (NF-κB), CX3CL1, and MMPs in human bronchial epithelial cells. Human bronchial epithelial cells were seeded in six-well plates. After 24 h, the cells were classified into six groups: control blank (CK) group; LPS group; PD98059 (ERK inhibitor) (PD) group, treated with LPS + ERK inhibitor; LY294002(PI3K/Akt inhibitor) (LY) group, treated with LPS + PI3K/Akt inhibitor; Aspirin (Asp) group, treated with LPS + aspirin; and Pyrrolidinedithiocarbamic acid (PDTC) group, treated with LPS + NF-κB inhibitor. After 4-h treatment, the cells were harvested. Western blot analysis was performed to detect the expression of PI3K/Akt, ERK, NF-κB, and CX3CL1. Real-time quantitative PCR (RT-qPCR) was used to determine the gene expression of MMP-7, MMP-9, and MMP-12. Compared to the CK group, expression of PI3K/Akt, ERK, NF-κB, and CX3CL1 was significantly increased in the LPS group (P < 0.05). When compared to the LPS group, expression of PI3K/Akt, ERK, NF-κB, and CX3CL1 was significantly decreased in the PD group, PDTC group, and Asp group (P < 0.05). In addition, expression of NF-κB in the LY group was significantly reduced by comparison with the LPS group (P < 0.05). RT-qPCR: When compared to the LPS group, expression of MMP-7 and MMP-12 was significantly decreased in Asp group (P < 0.05). Expression of MMP-12 was significantly reduced in LY group (P < 0.05). LPS-ERK, NF-κB-PI3K/Akt, and CX3CL1 signal pathways exist in human bronchial epithelial cells. The PI3K/Akt inhibitor repressed expression of MMP-12. Aspirin inhibited LPS-induced expression of PI3K, Akt, ERK, NF-κB, CX3CL1, MMP-7, and MMP-12 in human bronchial epithelial cells.
Similar content being viewed by others
References
Wang, L.C., J.N. Wu, W. Zhang, Y.H. Zhi, Y.C. Wu, R.L. Jiang, et al. 2013. Effects of aspirin on the ERK and PI3K/Akt signaling pathways in rats with acute pulmonary embolism. Molecular Medicine Reports 8: 1465–1471.
Wang, L.C., J.N. Wu, G.L. Xia, W. Mao, R.B. Ying, L.Q. Huang, et al. 2014. Effect of aspirin on fractalkine in rats with pulmonary embolism. Tropical Journal of Pharmaceutical Research 13: 753–760.
Wang, L.C., R.L. Jiang, W. Zhang, L.L. Wei, and R.H. Yang. 2014. Effects of aspirin on the expression of nuclear factor-kappaB in a rat model of acute pulmonary embolism. World Journal of Emergency Medicine 5: 229–233.
Li, Y., L. Zeng, and S. Liu. 2014. Total flavonoids from Plumula Nelumbinis suppress angiotensin II-induced fractalkine production by inhibiting the ROS/NF-kappa B pathway in human umbilical vein endothelial cells. Experimental Therapeutic Medicine 7: 1187–1192.
Slovak, M.L., V. Bedell, Y.H. Hsu, D.B. Estrine, N.J. Nowak, M.L. Delioukina, et al. 2011. Molecular karyotypes of Hodgkin and Reed-Sternberg cells at disease onset reveal distinct copy number alterations in chemosensitive versus refractory Hodgkin lymphoma. Clinical Cancer Research : an Official Journal of the American Association for Cancer Research 17: 3443–3454.
Ishida, Y., T. Hayashi, T. Goto, A. Kimura, S. Akimoto, N. Mukaida, et al. 2008. Essential involvement of CX3CR1-mediated signals in the bactericidal host defense during septic peritonitis. Journal of Immunology 181: 4208–4218.
Zhang, J., W. Yang, B. Hu, W. Wu, and M.B. Fallon. 2014. Endothelin-1 activation of the endothelin B receptor modulates pulmonary endothelial CX3CL1 and contributes to pulmonary angiogenesis in experimental hepatopulmonary syndrome. The American Journal of Pathology 184: 1706–1714.
Uchida, M., T. Ito, T. Nakamura, H. Igarashi, T. Oono, N. Fujimori, et al. 2013. ERK pathway and sheddases play an essential role in ethanol-induced CX3CL1 release in pancreatic stellate cells. Laboratory investigation. A Journal of Technical Methods and Pathology 93: 41–53.
Inui, M., Y. Ishida, A. Kimura, Y. Kuninaka, N. Mukaida, and T. Kondo. 2011. Protective roles of CX3CR1-mediated signals in toxin A-induced enteritis through the induction of heme oxygenase-1 expression. Journal of Immunology 186: 423–431.
Volin, M.V., N. Huynh, K. Klosowska, R.D. Reyes, and J.M. Woods. 2010. Fractalkine-induced endothelial cell migration requires MAP kinase signaling. Pathobiology: Journal of Immunopathology, Molecular and Cellular Biology 77: 7–16.
Muhl, D., S. Ghosh, J.A. Uzuelli, J. Lantos, and J.E. Tanus-Santos. 2010. Increases in circulating matrix metalloproteinase-9 levels following fibrinolysis for acute pulmonary embolism. Thrombosis Research 125: 549–553.
Voigt, P., C. Brock, B. Nurnberg, and M. Schaefer. 2005. Assigning functional domains within the p101 regulatory subunit of phosphoinositide 3-kinase gamma. The Journal of Biological Chemistry 280: 5121–5127.
Lin, C.H., S.H. Yeh, C.H. Lin, K.T. Lu, T.H. Leu, W.C. Chang, et al. 2001. A role for the PI-3 kinase signaling pathway in fear conditioning and synaptic plasticity in the amygdala. Neuron 31: 841–851.
Eblen, S.T., J.K. Slack, M.J. Weber, and A.D. Catling. 2002. Rac-PAK signaling stimulates extracellular signal-regulated kinase (ERK) activation by regulating formation of MEK1-ERK complexes. Molecular and Cellular Biology 22: 6023–6033.
Bazan, J.F., K.B. Bacon, G. Hardiman, W. Wang, K. Soo, D. Rossi, et al. 1997. A new class of membrane-bound chemokine with a CX3C motif. Nature 385: 640–644.
Jiang, J.X., Y. Zhang, S.H. Ji, P. Zhu, and Z.G. Wang. 2002. Kinetics of mitogen-activated protein kinase family in lipopolysaccharide-stimulated mouse Kupffer cells and their role in cytokine production. Shock 18: 336–341.
Shi, L., R. Kishore, M.R. McMullen, and L.E. Nagy. 2002. Lipopolysaccharide stimulation of ERK1/2 increases TNF-alpha production via Egr-1. American journal of physiology. Cell Physiology 282: C1205–1211.
Chen, T., Z.P. Guo, X.Y. Jiao, R.Z. Jia, Y.H. Zhang, J.Y. Li, et al. 2011. CCL5, CXCL16, and CX3CL1 are associated with Henoch-Schonlein purpura. Archives of Dermatological Research 303: 715–725.
Chen, T., Z.P. Guo, X.Y. Jiao, R.Z. Jia, Y.H. Zhang, J.Y. Li, et al. 2011. Peoniflorin suppresses tumor necrosis factor-alpha induced chemokine production in human dermal microvascular endothelial cells by blocking nuclear factor-kappaB and ERK pathway. Archives of Dermatological Research 303: 351–360.
van der Vorst, E.P., L.Z. Vanags, L.L. Dunn, H.C. Prosser, K.A. Rye, and C.A. Bursill. 2013. High-density lipoproteins suppress chemokine expression and proliferation in human vascular smooth muscle cells. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology 27: 1413–1425.
Bhattacharyya, S., S. Ghosh, and P.C. Sil. 2014. Amelioration of aspirin induced oxidative impairment and apoptotic cell death by a novel antioxidant protein molecule isolated from the herb Phyllanthus niruri. PloS One 9, e89026.
Sheu, M.L., F.M. Ho, K.F. Chao, M.L. Kuo, and S.H. Liu. 2004. Activation of phosphoinositide 3-kinase in response to high glucose leads to regulation of reactive oxygen species-related nuclear factor-kappaB activation and cyclooxygenase-2 expression in mesangial cells. Molecular Pharmacology 66: 187–196.
Chen, J., S. Zuo, J. Wang, J. Huang, X. Zhang, Y. Liu, et al. 2014. Aspirin promotes oligodendrocyte precursor cell proliferation and differentiation after white matter lesion. Frontiers in Aging Neuroscience 6: 7.
Zhang, F., M. Lu, H. Wang, and T. Ren. 2013. Aspirin attenuates angiotensin II-induced inflammation in bone marrow mesenchymal stem cells via the inhibition of ERK1/2 and NF-kappaB activation. Biomedical Reports 1: 930–934.
Stark, L.A., F.V. Din, R.M. Zwacka, and M.G. Dunlop. 2001. Aspirin-induced activation of the NF-kappaB signaling pathway: a novel mechanism for aspirin-mediated apoptosis in colon cancer cells. FASEB Journal: Official Publication of the Federation of American Societies for Experimental Biology 15: 1273–1275.
Reel, B., G.B. Sala-Newby, W.C. Huang, and A.C. Newby. 2011. Diverse patterns of cyclooxygenase-independent metalloproteinase gene regulation in human monocytes. British Journal of Pharmacology 163: 1679–1690.
Pul, R., T. Kopadze, T. Skripuletz, E.V. Voss, B.C. Kieseier, and M. Stangel. 2009. Polyclonal immunoglobulins (IVIg) induce expression of MMP-9 in microglia. Journal of Neuroimmunology 217: 46–50.
Dean, R.A., J.H. Cox, C.L. Bellac, A. Doucet, A.E. Starr, and C.M. Overall. 2008. Macrophage-specific metalloelastase (MMP-12) truncates and inactivates ELR+ CXC chemokines and generates CCL2, -7, -8, and -13 antagonists: potential role of the macrophage in terminating polymorphonuclear leukocyte influx. Blood 112: 3455–3464.
Bode, W., and K. Maskos. 2003. Structural basis of the matrix metalloproteinases and their physiological inhibitors, the tissue inhibitors of metalloproteinases. Biological Chemistry 384: 863–872.
Williams, H., J.L. Johnson, C.L. Jackson, S.J. White, and S.J. George. 2010. MMP-7 mediates cleavage of N-cadherin and promotes smooth muscle cell apoptosis. Cardiovascular Research 87: 137–146.
Eldrup, N., M.L. Gronholdt, H. Sillesen, and B.G. Nordestgaard. 2006. Elevated matrix metalloproteinase-9 associated with stroke or cardiovascular death in patients with carotid stenosis. Circulation 114: 1847–1854.
Cau, S.B., R.C. Barato, M.R. Celes, J.J. Muniz, M.A. Rossi, and J.E. Tanus-Santos. 2013. Doxycycline prevents acute pulmonary embolism-induced mortality and right ventricular deformation in rats. Cardiovascular Drugs and Therapy / Sponsored by the International Society of Cardiovascular Pharmacotherapy 27: 259–267.
Werb, Z., and S. Gordon. 1975. Elastase secretion by stimulated macrophages. Characterization and regulation. The Journal of Experimental Medicine 142: 361–377.
Shapiro, S.D., D.K. Kobayashi, and T.J. Ley. 1993. Cloning and characterization of a unique elastolytic metalloproteinase produced by human alveolar macrophages. The Journal of Biological Chemistry 268: 23824–23829.
Lu, L., H. Liu, J. Peng, L. Gan, L. Shen, Q. Zhang, et al. 2010. Regulations of the key mediators in inflammation and atherosclerosis by aspirin in human macrophages. Lipids in Health and Disease 9: 16.
Hua, Y., J. Xue, F. Sun, L. Zhu, and M. Xie. 2009. Aspirin inhibits MMP-2 and MMP-9 expressions and activities through upregulation of PPARalpha/gamma and TIMP gene expressions in ox-LDL-stimulated macrophages derived from human monocytes. Pharmacology 83: 18–25.
Yang, X.S., S.A. Liu, J.W. Liu, and Q. Yan. 2012. Fucosyltransferase IV enhances expression of MMP-12 stimulated by EGF via the ERK1/2, p38 and NF-kappaB pathways in A431 cells. Asian Pacific Journal of Cancer Prevention : APJCP 13: 1657–1662.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Funding
Funded by Zhejiang Provincial Program for the Cultivation of High-Level Innovative Health Talents (2014-18), the medical and health platform program (key support) of Zhejiang province (Grant No. 2015ZDA022), the Natural Sciences Fund of Zhejiang Province (Grant No. LY12H29005), and Key Disciplines of Integrative Traditional Chinese Medicine and Western Medicine of Zhejiang Province (Grant No. 2012-XK-A12).
Conflict of Interest
We have no conflicts of interest to declare.
Rights and permissions
About this article
Cite this article
Jiang, R., Wei, L., Zhu, M. et al. Aspirin Inhibits LPS-Induced Expression of PI3K/Akt, ERK, NF-κB, CX3CL1, and MMPs in Human Bronchial Epithelial Cells. Inflammation 39, 643–650 (2016). https://doi.org/10.1007/s10753-015-0289-8
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10753-015-0289-8