Daily in vitro dosing of conjugated linoleic acid on THP-1 cultured monocytes
Cardiff Metropolitan University
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BACKGROUND: Conjugated linoleic acid (CLA) has been established as a natural peroxisome proliferator activated receptor (PPAR-y) ligand. Recent studies have explored the possible health benefits associated with CLA for the treatment and prevention of type 2 diabetes (T2D) in relation to the insulin sensitising properties of PPAR-y activation. However, synthetic PPAR-y ligands such as rosiglitazone are claimed to exert detrimental cardiovascular complications. Thus, with the alarming side effects possibly generated from rosiglitazone, this study will test the hypothesis that natural PPAR-y ligand CLA does not exert cytotoxic effects on monocytic cell viability. METHOD: Cell culture techniques employed THP-I monocytic cells. THP-I monocytes were treated with CLA (20µM and 100µM), once with a bolus dose and with a repeated dose which was administered at every passage (3/4 days) over a 14 day duration. Caspase glo 3/7 and MTS assays were preformed daily to assess cell viability and apoptosis. RESULTS: As expected, the cytotoxin thapsigargin significantly reduced cell viability and increased apoptosis, coinciding with previous research. Moreover, data illustrated a significant difference between CLA treated and non-treated cells (P=0.008, CI 99.34%), suggesting CLA has a significant influence on reducing cell viability and increasing cellular apoptosis. Specifically, cells treated with CLA 100µM repeatedly over the 14 days yielded a significant increase in apoptosis and decrease in viable cell amounts (P=0.02). CONCLUSION: CLA exerted a significant cytotoxic effect on THP-I monocytic cells in comparison to non-cytotoxic outcomes of rosiglitazone and pioglitazone treated cells from the same research group. Data presented in this investigation suggests that endoplasmic reticulum (ER) stress is influential in initiating cellular apoptosis, thus supra-pharmacological CLA concentrations may be discouraged as a therapeutic treatment due to the cytotoxic cellular effects and possibly detrimental pathological effects in relation to T2D and its associated complications.
BSc (Hons) Sports Biomedicine and Nutrition
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