
ABSTRACT
Background and Objective: Both leukocyte differentiation antigen 14 (CD14) and myeloid differentiation protein-2 (MD-2) contain hydrophobic vesicles for extracting and delivering lipopolysaccharide (LPS), which play an irreplaceable role in the extracellular signal transduction of LPS. Chlorogenic acid (CGA) is a phenolic compound with a strong anti-inflammatory effect. However, it is currently unclear whether CGA blocks the extracellular signal transmission of LPS by occupying the hydrophobic pockets of CD14 and MD-2. Materials and Methods: Flow cytometry and siRNA were used to detect the presence of CD14 and MD-2 on breast epithelial cells (MECs). Molecular docking, Western blot, reverse transcription-polymerase chain reaction and enzyme-linked immunosorbent assay were used to study the targeting effect of CGA on CD14 and MD-2 and its influence on cytokine secretion. The protective effect of CGA on breast inflammation was observed under a transmission electron microscope. Results: The results showed that MD-2 and CD14 were expressed on MECs. When CD14 and MD-2 are inhibited alone or at the same time, the binding rate of LPS and MECs and the secretion of cytokines are reduced. CGA can not only occupy the hydrophobic vesicles of CD14 and MD-2, block the extracellular signal transmission of LPS but also inhibit the expression of CD14 and MD-2, thereby blocking the signal transmission of LPS from extracellular to intracellular. Conclusion: Besides, CGA can effectively inhibit the general changes of breast tissue, breast inflammatory cell infiltration, mitochondria and rough endoplasmic reticulum damage caused by lipopolysaccharide. In conclusion, CGA is expected to act as an antagonist of CD14 and MD-2 to inhibit mastitis.
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How to cite this article
JunChao Peng, YuKun Wang, XueYan Xie, Qiong Yi, Xin Li, YuHao Wei, XuHua He and Lu Wang, 2020. Chlorogenic Acid Inhibits LPS-Induced Mammary Epithelial Cell Inflammation in Mice by Targeting CD14 and MD-2. International Journal of Pharmacology, 16: 542-553.
DOI: 10.3923/ijp.2020.542.553
URL: https://scialert.net/abstract/?doi=ijp.2020.542.553
DOI: 10.3923/ijp.2020.542.553
URL: https://scialert.net/abstract/?doi=ijp.2020.542.553
REFERENCES
- Ulevitch, R.J. and P.S. Tobias, 1995. Receptor-dependent mechanisms of cell stimulation by bacterial endotoxin. J. Annu. Rev. Immunol., 13: 437-457.
CrossRefPubMedDirect Link - Kelley, S.L., T. Lukk, S.K. Nair and R.I. Tapping, 2013. The crystal structure of human soluble CD14 reveals a bent solenoid with a hydrophobic amino-terminal pocket. J.I. 190: 1304-1311.
CrossRefDirect Link - Kim, J.I., C.J. Lee, M.S. Jin, C.H. Lee, S.G. Paik, H. Lee and J.O. Lee, 2005. Crystal structure of CD14 and its implications for lipopolysaccharide signaling. J. Biol. Chem., 280: 11347-11351.
CrossRefDirect Link - Viriyakosol, S., P.S. Tobias, R.L. Kitchens and T.N. Kirkland, 2001. MD-2 binds to bacterial lipopolysaccharide. J. Biol. Chem., 276: 38044-38051.
CrossRefDirect Link - Schromm, A.B., E. Lien, P. Henneke, J.C. Chow and A. Yoshimura et al., 2001. Molecular genetic analysis of an endotoxin nonresponder mutant cell line: A point mutation in a conserved region of MD-2 abolishes endotoxin-induced signaling. J. Exp. Med., 194: 79-88.
CrossRefPubMedDirect Link - Rallabhandi, P., J. Bell, M.S. Boukhvalova, A. Medvedev and E. Lorenz et al., 2006. Analysis of TLR4 polymorphic variants: New insights into TLR4/MD-2/CD14 stoichiometry, structure and signaling. J. Immunol., 177: 322-332.
CrossRefDirect Link - Gay, N.J. and M. Gangloff, 2007. Structure and function of toll receptors and their Ligands. Ann. Rev. Biochem., 76: 141-165.
CrossRefDirect Link - Medzhitov, R., P. Preston-Hurlburt and C.A. Janeway, 1997. A human homologue of the Drosophila toll protein signals activation of adaptive immunity. Nature, 388: 394-397.
CrossRefDirect Link - Raetz, C.R. and C. Whitfield, 2002. Lipopolysaccharide endotoxins. Annu. Rev. Biochem., 71: 635-700.
CrossRefPubMedDirect Link - Park, B.S., D.H. Song, H.M. Kim, B.S. Choi, H. Lee and J.O. Lee, 2009. The structural basis of lipopolysaccharide recognition by the TLR4–MD-2 complex. Nature, 458: 1191-1195.
CrossRefDirect Link - Nagai, Y., S. Akashi, M. Nagafuku, M. Ogata and Y. Iwakura et al., 2002. Essential role of MD-2 in LPS responsiveness and TLR4 distribution. Nat. Immunol., 3: 667-672.
CrossRefDirect Link - Ohto, U., K. Fukase, K. Miyake and Y. Satow, 2007. Crystal structures of human MD-2 and its complex with antiendotoxic lipid IVa. Science, 316: 1632-1634.
CrossRefDirect Link - Resman, N., J. Vašl, A. Oblak, P. Pristovšek, T.L. Gioannini, J.P. Weiss and R. Jerala, 2009. Essential roles of hydrophobic residues in both MD-2 and toll-like receptor 4 in activation by endotoxin. J. Biol. Chem., 284: 15052-15060.
CrossRefDirect Link - Hebeda, C.B., S.M. Bolonheis, A. Nakasato, K. Belinati and P.D.C. Souza et al., 2011. Effects of chlorogenic acid on neutrophil locomotion functions in response to inflammatory stimulus. J. Ethnopharmacol., 135: 261-269.
CrossRef - Stoclet, J.C., T. Chataigneau, M. Ndiaye, M.H. Oak, J. El Bedoui, M. Chataigneau and V.B. Schini-Kerth 2004. Vascular protection by dietary polyphenols. Eur. J. Pharmacol., 500: 299-313.
CrossRefDirect Link - Olmos, A., R.M. Giner, M.C. Recio, J.L. Rios, J.M. Cerdá-Nicolás and S. Máñez, 2007. Effects of plant alkylphenols on cytokine production, tyrosine nitration and inflammatory damage in the efferent phase of contact hypersensitivity. Brit. J. Pharmacol., 152: 366-373.
CrossRefDirect Link - Zhang, X., H. Huang, T. Yang, Y. Ye, J. Shan, Z. Yin and L. Luo, 2010. Chlorogenic acid protects mice against lipopolysaccharide-induced acute lung injury. Injury, 41: 746-752.
CrossRefDirect Link - Xu, Y., J. Chen, X. Yu, W. Tao, F. Jiang, Z. Yin and C. Liu, 2010. Protective effects of chlorogenic acid on acute hepatotoxicity induced by lipopolysaccharide in mice. Inflamm. Res., 59: 871-877.
CrossRefDirect Link - Feng, R., Y. Lu, L.L. Bowman, Y. Qian, V. Castranova and M. Ding, 2005. Inhibition of activator protein-1, NF-κB and MAPKs and induction of phase 2 detoxifying enzyme activity by chlorogenic acid. J. Biol. Chem., 280: 27888-27895.
CrossRefDirect Link - Zheng, Z., L. Shi, Y. Sheng, J. Zhang, B. Lu and L. Ji, 2016. Chlorogenic acid suppresses monocrotaline-induced sinusoidal obstruction syndrome: The potential contribution of NFκB, Egr1, Nrf2, MAPKs and PI3K signals. Environ. Toxicol. Pharmacol., 46: 80-89.
CrossRefDirect Link - He, C.L., Q. Yi, Y.F. Li, H. Yang and L. Wang, 2013. Toll-like receptor-4, but not toll-like receptor-2 mediates secretion of tumour necrosis factor α and interleukin-8 in lipopolysaccharide-stimulated mouse mammary epithelial cells. Bull. Vet. Inst. Pulawy, 57: 393-397.
CrossRefDirect Link - Alper, S., R. Laws, B. Lackford, W.A. Boyd, P. Dunlap, J.H. Freedman and D.A. Schwartz, 2008. Identification of innate immunity genes and pathways using a comparative genomics approach. Proc. Nat. Acad. Sci., 105: 7016-7021.
CrossRefDirect Link - Sathishkumar, N., S. Sathiyamoorthy, M. Ramya, D.U. Yang, H.N. Lee and D.C. Yang, 2012. Molecular docking studies of anti-apoptotic BCL-2, BCL-XL and MCL-1 proteins with ginsenosides from Panax ginseng. J.Enz. Inhib. Med. Chem., 27: 685-692.
CrossRefDirect Link - Shishodia, S., S. Majumdar, S. Banerjee and B.B. Aggarwal, 2003. Ursolic acid inhibits nuclear factor-kappaB activation induced by carcinogenic agents through suppression of IkappaBalpha kinase and p65 phosphorylation: Correlation with down-regulation of cyclooxygenase 2, matrix metalloproteinase 9 and cyclin D1. Cancer Res., 63: 4375-4383.
PubMed - Mosmann, T., 1983. Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity assays. J. Immunol. Methods, 65: 55-63.
CrossRefPubMedDirect Link - Pfaffl, M.W. and M. Hageleit, 2001. Validities of mRNA quantification using recombinant RNA and recombinant DNA external calibration curves in real-time RT-PCR. Biotech. Lett., 23: 275-282.
CrossRef - Zhang, X., Y. Wang, C. Xiao, Z. Wei, J. Wang, Z. Yang and Y. Fu, 2017. Resveratrol inhibits LPS-induced mice mastitis through attenuating the MAPK and NF-κB signaling pathway. Microb. Pathogen., 107: 462-467.
CrossRefDirect Link - Haziot, A., E. Ferrero, F. Köntgen, N. Hijiya and S. Yamamoto et al., 1996. Resistance to endotoxin shock and reduced dissemination of gram-negative bacteria in CD14-deficient mice. Immunity, 4: 407-414.
CrossRefPubMedDirect Link - Juan, T.S.C., E. Hailman, M.J. Kelley, L.A. Busse and E. Davy et al., 1995. Identification of a lipopolysaccharide binding domain in CD14 between amino acids 57 and 64. J. Biol. Chem., 270: 5219-5224.
CrossRefDirect Link - Stelter, F., M. Bernheiden, R. Menzel, R.S. Jack, S. Witt, X. Fan, M. Pfister and C. Schutt, 1997. Mutation of amino acids 39-44 of human CD14 abrogates binding of lipopolysaccharide and Escherichia coli. Eur. J. Biochem., 243: 100-109.
CrossRefDirect Link - Morizot, B., 2012. Chance: From metaphysical principle to explanatory concept. The idea of uncertainty in a natural history of knowledge. Prog. Biophys. Mol. Biol., 110: 54-60.
CrossRefDirect Link