Shastri, MD and Stewart, N and Horne, J and Peterson, GM and Gueven, N and Sohal, SS and Patel, RP, In-vitro suppression of IL-6 and IL-8 release from human pulmonary epithelial cells by non-anticoagulant fraction of enoxaparin, PLoS One, 10, (5) Article 0126763. ISSN 1932-6203 (2015) [Refereed Article]
© 2015 Shastri et al. Licensed under Creative Commons Attribution 4.0 International (CC BY 4.0) http://creativecommons.org/licenses/by/4.0/
Background: Enoxaparin, a mixture of anticoagulant and non-anticoagulant fractions, is widely used as an anticoagulant agent. However, it is also reported to possess anti-inflammatory properties. Our study indicated that enoxaparin inhibits the release of IL-6 and IL-8 from A549 pulmonary epithelial cells. Their release causes extensive lung tissue damage. The use of enoxaparin as an anti-inflammatory agent is hampered due to the risk of bleeding associated with its anticoagulant fractions. Therefore, we aimed to identify the fraction responsible for the observed anti-inflammatory effect of enoxaparin and to determine the relationship between its structure and biological activities.
Methods: A549 pulmonary epithelial cells were pre-treated in the presence of enoxaparin and its fractions. The levels of IL-6 and IL-8 released from the trypsin-stimulated cells were measured by ELISA. The anticoagulant activity of the fraction responsible for the effect of enoxaparin was determined using an anti-factor-Xa assay. The fraction was structurally characterised using nuclear magnetic resonance. The fraction was 2-O, 6-O or N-desulfated to determine the position of sulfate groups required for the inhibition of interleukins. High-performance size-exclusion chromatography was performed to rule out that the observed effect was due to the interaction between the fraction and trypsin or interleukins.
Results: Enoxaparin (60μg/mL) inhibited the release of IL-6 and IL-8 by >30%. The fraction responsible for this effect of enoxaparin was found to be a disaccharide composed of α-L-iduronic-acid and α-D-glucosamine-6-sulfate. It (15μg/mL) inhibited the release of interleukins by >70%. The 6-O sulphate groups were responsible for its anti-inflammatory effect. The fraction did not bind to trypsin or interleukins, suggesting the effect was not due to an artefact of the experimental model.
Conclusion: The identified disaccharide has no anticoagulant activity and therefore eliminates the risk of bleeding associated with enoxaparin. Future in-vivo studies should be designed to validate findings of the current study.
|Item Type:||Refereed Article|
|Keywords:||heparin, non anticoagulant, chromatography|
|Research Division:||Biomedical and Clinical Sciences|
|Research Group:||Pharmacology and pharmaceutical sciences|
|Research Field:||Pharmaceutical sciences|
|Objective Group:||Public health (excl. specific population health)|
|Objective Field:||Public health (excl. specific population health) not elsewhere classified|
|UTAS Author:||Shastri, MD (Mr Madhur Shastri)|
|UTAS Author:||Stewart, N (Dr Niall Stewart)|
|UTAS Author:||Horne, J (Dr James Horne)|
|UTAS Author:||Peterson, GM (Professor Gregory Peterson)|
|UTAS Author:||Gueven, N (Dr Nuri Guven)|
|UTAS Author:||Sohal, SS (Dr Sukhwinder Sohal)|
|UTAS Author:||Patel, RP (Dr Rahul Patel)|
|Web of Science® Times Cited:||29|
|Downloads:||424 View Download Statistics|
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