Differential effects of performance-enhancing drugs 'Methamphetamine' and 'hCG' on ex-vivo cultured primary blood mononuclear cells of male athletes.

BACKGROUND

To bridge the knowledge gap, the present study aimed to investigate the effect of different doses of two widely used performance-enhancing drugs 'methamphetamine' (Meth) and 'human chorionic gonadotropin'(hCG) on ex-vivo cultured primary blood cells of young male Indian athletes.

METHODS

Primary blood mononuclear cells (PBMCs) were isolated and cultured to obtain pure T cells and monocyte-derived macrophages. Immunofluorescence, flow-cytometry, qRT-PCR, Western blot, ELISA and siRNA transfection studies were carried out to evaluate the effect of these two drugs on athletes' blood cells.

RESULTS

Cell viability studies revealed that Meth at high doses was toxic for PBMCs and showed a significant negative impact on red blood cell fragility but hCG incubation did not result in any cytotoxicity or haemolysis. The current study also demonstrated that Meth incubation significantly affected T cell proliferation, percentage of regulatory T cells (Treg cells), Th17 cells, early activated T cells, ROS generation and mitochondrial dysfunction. On the other hand, hCG treatment upregulated the percentage of Treg cells. Within macrophage cells, Meth incubation upregulated MYD88 dependent TLR4 pathway and decreased the phagocytotic capability of the cells. Both hCG and Meth showed its potential action on alteration the pro/anti-inflammatory cytokine profiling but suppression of TLR4 pathway by RNA interference (TLR4 siRNA) suggested promising future treatment modalities.

CONCLUSION

This study demonstrated the differential effects of Meth and hCG on immune cells of athlete's blood. Meth acted as an inflammation and T cell dysfunction inducing agent, while hCG acted as an anti-inflammatory immunosuppressive molecule.