Opposing effects of low molecular weight heparins on the release of inflammatory cytokines from peripheral blood mononuclear cells of asthmatics.
BACKGROUND
T-cell-mediated inflammatory cytokines, such as interleukin (IL)-4, IL-5, IL-13 and tumor necrosis factor-alpha (TNF-α), play an important role in the initiation and progression of inflammatory airways diseases. Low-molecular-weight heparins (LMWHs), widely used anticoagulants, possess anti-inflammatory properties making them potential treatment options for inflammatory diseases, including asthma. In the current study, we investigated the modulating effects of two LMWHs (enoxaparin and dalteparin) on the release of cytokines from stimulated peripheral blood mononuclear cells (PBMCs) of asthmatic subjects to identify the specific components responsible for the effects.


METHODS
PBMCs from asthmatic subjects (consist of ~75% of T-cells) were isolated from blood taken from ten asthmatic subjects. The PBMCs were pre-treated in the presence or absence of different concentrations of LMWHs, and were then stimulated by phytohaemagglutinin for the release of IL-4, IL-5, IL-13 and TNF-α. LMWHs were completely or selectively desulfated and their anticoagulant effect, as well as the ability to modulate cytokine release, was determined. LMWHs were chromatographically fractionated and each fraction was tested for molecular weight determination along with an assessment of anticoagulant potency and effect on cytokine release.


RESULTS
Enoxaparin inhibited cytokine release by more than 48%, whereas dalteparin increased their release by more than 25%. The observed anti-inflammatory effects of enoxaparin were independent of their anticoagulant activities. Smaller fractions, in particular dp4 (four saccharide units), were responsible for the inhibitory effect of enoxaparin. Whereas, the larger fractions, in particular dp22 (twenty two saccharide units), were associated with the stimulatory effect of dalteparin.


CONCLUSIONS
Enoxaparin and dalteparin demonstrated opposing effects on inflammatory markers. These observed effects could be due to the presence of structurally different components in the two LMWHs arising from different methods of depolymerisation. This study provides a platform for further studies investigating the usefulness of enoxaparin in various inflammatory diseases.
School of Medicine, Division of Pharmacy, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Madhur D
MD
Shastri
School of Medicine, Division of Pharmacy, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Niall
N
Stewart
Breathe Well Centre of Research Excellence for Chronic Respiratory Disease and Lung Ageing, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Mathew
M
Eapen
School of Medicine, Division of Pharmacy, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia; Breathe Well Centre of Research Excellence for Chronic Respiratory Disease and Lung Ageing, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Gregory M
GM
Peterson
School of Medicine, Division of Pharmacy, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Syed Tabish R
ST
Zaidi
School of Medicine, Division of Pharmacy, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Nuri
N
Gueven
Breathe Well Centre of Research Excellence for Chronic Respiratory Disease and Lung Ageing, School of Medicine, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Sukhwinder Singh
SS
Sohal
School of Medicine, Division of Pharmacy, Faculty of Health, University of Tasmania, Hobart, Tasmania, Australia.
Rahul P
RP
Patel
D000328:Adult; D000893:Anti-Inflammatory Agents / Q000737:chemistry / Q000494:pharmacology*; D001249:Asthma / Q000276:immunology*; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D016207:Cytokines / Q000378:metabolism*; D005260:Female; D006495:Heparin, Low-Molecular-Weight / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D008297:Male; D008875:Middle Aged; D009844:Oligosaccharides / Q000032:analysis / Q000302:isolation & purification; D013431:Sulfates / Q000737:chemistry
D000893:Anti-Inflammatory Agents; D016207:Cytokines; D006495:Heparin, Low-Molecular-Weight; D009844:Oligosaccharides; D013431:Sulfates