Rats were divided into four groups of eight animals each. Group I rats were control rats; injected with saline (CT), group II rats were injected with saline and treated with chlorogenic acid (AC), group III rats were injected with 0.1% ethidium bromide (EB) and group IV rats were injected with 0.1% EB and treated with chlorogenic acid (EB + AC). The activities of the enzymes were analyzed using colorimetric methods, and the gene expression of NTPDase 1, 2 and 3 were analyzed using the polymerase chain reaction (PCR).
The results revealed that there was a significant (P < 0.01) reduction in E-NPP activity in EB group (1.63 ± 0.10 nmol p-nitrophenol released/min/mg protein) when compared to CT group (2.33 ± 0.14 nmol p-nitrophenol released/min/mg protein). However, treatment with chlorogenic acid significantly (P < 0.05) increased E-NPP activity in EB group. Furthermore, no significant (P > 0.05) change was observed in the E-NPP activity of EB + AC group (2.19 ± 0.08 nmol p-nitrophenol released/min/mg protein) when compared to CT group (2.33 ± 0.14 nmol p-nitrophenol released/min/mg protein). In addition, there was a significant (P < 0.05) increase in AMP hydrolysis in EB rat group when compared to CT group. No significant (P > 0.05) difference was observed in AMP hydrolysis between AC, AC + EB and CT groups. Conversely, there were no significant (P > 0.05) differences in ATP and ADP hydrolyses between all the groups (AC, EB, AC + EB and CT groups). Likewise, there were no significant (P > 0.05) changes in E-ADA activity and percentage platelet aggregation among all groups studied. Similarly, no significant (P > 0.05) change was observed in the expression of E-NTPDase 1, 2 and 3 in all the groups tested.
Our study revealed that chlorogenic acid may modulate the hydrolysis of adenine nucleotides in platelets of rats demyelinated and treated with chlorogenic acid via alteration of E-NPP and ecto-5′-nucleotidase activities.