Thesis

Control of cytokine production from human monocytic cells by prostaglandin 15-Deoxy-Δ-12,14-prostaglandin J2

Creator
Awarding institution
  • University of Strathclyde
Date of award
  • 2018
Thesis identifier
  • T15284
Person Identifier (Local)
  • 201753568
Qualification Level
Qualification Name
Department, School or Faculty
Abstract
  • Prostaglandins have several roles, but their main function is within the inflammatory process. As part of the immune response, immune cells are stimulated by lipopolysaccharide to produce pro- and anti-inflammatory cytokines including tumour necrosis factor-alpha, interleukin-1beta and interleukin-10. These cytokines can go on to stimulate the production of prostaglandins such as prostaglandin E2, prostaglandin D2 and 15-deoxy-Δ-12,14-prostaglandin J2. 15-deoxy-Δ-12,14-prostaglandin J2, is a metabolite of prostaglandin D2.;Prostaglandins have both pro- and anti-inflammatory roles. Not much is known about the pro- and anti-inflammatory effects of 15-deoxy-Δ-12,14-prostaglandin J2. As prostaglandins like prostaglandin E2, have inhibitory effects on pro-inflammatory cytokines it is an intriguing possibility that 15-deoxy-Δ-12,14-prostaglandin J2 may also share these qualities. It is also not known what cell receptor mediates these responses or what intracellular pathway is activated.;Thus, our aim was to discover what effects that 15-deoxy-Δ-12,14 -prostaglandin J2 had on the levels of pro- and anti-inflammatory cytokines, tumour necrosis factor-alpha, interleukin-1beta and interleukin-10. As well as to understand what receptor, either the prostaglandin D2 receptors DP1 or DP2, that 15-deoxy-Δ-12,14-prostaglandin J2 binds to and if cyclic adenosine monophosphate is involved in the signalling pathway.;This was achieved by stimulating the human monocytic cell line, THP-1 cells, with lipopolysaccharide and incubated for 22 hours with prostaglandins; 15-deoxy-Δ-12,14-prostaglandin J2, prostaglandin D2 and prostaglandin E2 which were used as comparisons. DP1 and DP2 antagonists BW868C and CAY10471 respectively and the cyclic adenosine monophosphate inhibitor SQ22536. The supernatants which contained the cytokines were measured by enzyme-linked immunosorbent assay. After the incubations, the cells were removed, and cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay after stimulation with lipopolysaccharide in the presence or absence of prostaglandin E2.;15-deoxy-Δ-12,14-prostaglandin J2 downregulated the levels of tumour necrosis factor-alpha by 81% and interleukin-1beta by 33% in a concentration dependent manner but had no effect on the levels of interleukin-10. As the concentration of prostaglandin E2 increased it downregulated the levels of tumour necrosis factor-alpha, interleukin-1beta and interleukin-10 by 100%, 69% and 82.6% respectively.;Surprisingly, neither of the DP antagonists had any effect on the activity of 15-deoxy-Δ-12,14-prostaglandin J2. CAY10471 (DP2), inhibited prostaglandin D2 from decreasing the level of tumour necrosis factor-alpha, but BW868C (DP1) did not have any effect. When using the cyclic adenosine monophosphate inhibitor, SQ22536, it had no effect on the levels of tumour necrosis factor-alpha, with either 15-deoxy-Δ-12,14-prostaglandin J2 or prostaglandin D2. Lipopolysaccharide and prostaglandin E2 had no effect on THP-1 cell viability confirmed by no change in the converted formazan dye.;This data indicates that 15-deoxy-Δ-12,14-prostaglandin J2 has inhibitory effects on pro-inflammatory cytokines tumour necrosis factor-alpha and interleukin -1beta but has no effect on the anti-inflammatory cytokine interleukin-10 in human monocytic cells. The data strongly suggests that as the DP antagonists BW868C and CAY10471 do not affect the function of 15-deoxy-Δ-12,14-prostaglandin J2 then neither the DP1 or DP2 receptors are involved in the anti-inflammatory effects of 15-deoxy-Δ-12,14-prostaglandin J2.;Also, as the cyclic adenosine monophosphate inhibitor SQ22536 had no effect either on the function of 15-deoxy-Δ-12,14-prostaglandin J2, this rules out that the cyclic adenosine monophosphate pathway is involved. This is inclusive of any other receptors that follow this pathway. Further work can be carried out to understand what receptor and pathway are stimulated. As 15-deoxy-Δ-12,14-prostaglandin J2 has shown to be suppressive of tumour necrosis factor-alpha and interleukin -1beta, and as it does not alter the levels of the anti-inflammatory cytokine interleukin-10, it is propitious that it may be used successfully to modulate the inflammatory process and be developed into a therapy for inflammatory conditions.
Advisor / supervisor
  • Rotondo, Dino
Resource Type
DOI
Date Created
  • 2018
Former identifier
  • 9912728482402996

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