AAGP™ has an Additive Protective Effect on Oxidative Stress Induced Cell Death, in the Presence of Coenzyme Q10, in Human Adult and Neonatal Fibroblasts at 37°C

Hussein, January 2006

Introduction

Previous results demonstrated AAGP™ protective activity against H2O2 exposure and UV-C radiation is still present and strong at low concentrations. In the following experiments, AAGP™ will be compared and added in combination with other protective agents, such Coenzyme Q10, a strong anti-oxidant, to see whether or not AAGP™ will show additive properties to known protective agents already in use.

Method

Primary fetal and adult fibroblasts were plated at a concentration of 1x105 cells/ml and grown on non-adherent plates in serum-free media at 37°C with concentrations of either 0, 1 or 5mg/ml AAGP™, with 10nM Coenzyme Q10. To test the affects of these two molecules on oxidative stress the cells were either exposed to 1mM hydrogen peroxide or UVC (256nm) rays for 3 hours. Cell counts were taken at various time points using Trypan blue and two counts were performed per sample to calculate the percentage viability (from 0 hours) of the cell sample.

Results

As can be seen from the graphs AAGP™ and Coenzyme Q10 together provide more protection against UV and hydrogen peroxide induced oxidative stress than either one alone. These results demonstrate that combining AAGP™ with CoQ10 had an additive effect. AAGP™ synergies with CoQ10 to obtain increased cellular protection from UV or hydrogen peroxide induced oxidative stress.

Conclusion

CoQ10 is a potent antioxidant found in the cells of the body which is now being added to some skin care products as an anti-aging substance and to prevent oxidative damage caused by environmental stresses. The additive effect of AAGP™ and CoQ10 suggest AAGP™ doesn't have the same mechanism of action as CoQ10, which basically acts as a free radical scavenger.

One possible mechanism is that AAGP™ alters the extracellular pH and in turn helps the cells to cope with stressful environments. In our previous experiments, we showed that AAGP™ changes the pH to a more acidic pH, but in the absence of AAGP™, simply changing the pH was not enough to promote cell survival to the same extent, suggesting that pH change is not the only way AAGP™ could protect cells. Another possibility, which is affected by both pH alteration and Anti-freeze proteins, is control of Ca+ influx; Ca+ accumulation is detrimental to cells and organs, and both pH change and anti-freeze proteins have been shown to inhibit this event. More future experiments will need to be conducted to address the mechanism of action of AAGP™.