The effect of conjugated linoleic acid on the expression of the receptor for advanced glycated end-products (RAGE) in monocytic cells.
Cardiff Metropolitan University
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The receptor for advanced glycated end-product (RAGE) is recognised as a pattern recognition receptor capable of biochemical interactions with diverse endogenous ligands. RAGE is thought to be involved in the many complications of diabetes mellitus. The expression of RAGE is regulated by NF-κB transcription factors, the activation of RAGE-NF-κB signalling route produces an increased cell surface expression of RAGE, subsequently resulting in a positive-feedback mechanism that intensifies the initial signal and enhances the inflammation. It has been identified that the trans-10 cis-12 conjugated linoleic acid (CLA) isomer is capable of inhibition of NF-κB but also other inflammatory mechanisms in p65 and p50 subunits through prevention of their binding to DNA. Plus Marx et al., highlighted that a commonly used anti-diabetic agent thiazolidinediones (TZDs) are capable to down-regulate RAGE mRNA levels through the inhibition of NF-kB activation at the proximal NF-kB site of the RAGE promoter The optimisation of quantitative PCR was achieved indicating that iQ SYBR Green is the valid choice for running PCR (Relative Fluorescence Units 1300 +/- 100, 800 +/- 50, 200 +/- 75, iQ SYBR Green v FAST SYBR Green v Power SYBR Green). The second part of this investigation is designed to examine the regulation of RAGE gene expression in human THP-1 cell. THP-1 cells were exposed to conjugated linoleic acid (CLA) for 24hours, prior to amplification of the RAGE gene and subsequent quantitative polymerase chain reaction (Q- PCR), to derive the effect of CLA on RAGE expression. The data demonstrated that CLA reduced RAGE gene expression (delta Ct 1 +/- 0.1 vs 0.5 +/- 0.3 , control v CLA) results from Fig 3.15) although this down-regulation was non statistically significant (T-Test, P- value of 0.107). Nevertheless there are still areas of this work that can produce future prospect studies through; an analysis on longer exposure time, higher dosages of CLA or targeting different tissue cultures.
MSc Biomedical Sciences
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