Effects of chlorogenic acid on adenine nucleotides hydrolyzing enzyme activities and expression in platelets of rats experimentally demyelinated with ethidium bromide.
BACKGROUND
The effects of chlorogenic acid (one of the major phenolic acid found in human diets) were investigated on the adenine nucleotides hydrolyzing enzymes; ecto-nucleotide pyrophosphatase/phophodiesterase (E-NPP), ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase), E-5'- nucleotidase and ecto-adenosine deaminase (E-ADA) activities and expression in platelets of rats experimentally demyelinated with ethidium bromide.


METHODS
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).


RESULTS
The results revealed that there was a significant (P<0.01) reduction in E-NPP activity in EB group (1.63±0.10nmol p-nitrophenol released/min/mg protein) when compared to CT group (2.33±0.14nmol 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±0.08nmol p-nitrophenol released/min/mg protein) when compared to CT group (2.33±0.14nmol 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.


CONCLUSIONS
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.
Departament of Biochemistry and Molecular Biology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil. Electronic address: camleal@terra.com.br.
Claudio A M
CAM
Leal
Departament of Microbiology and Parasitology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Daniela B R
DBR
Leal
Departament of Microbiology and Parasitology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil; Department of Biochemistry, Federal University of Technology, P. M. B. 704, Akure 340001, Nigeria.
Stephen A
SA
Adefegha
Departament of Biochemistry and Molecular Biology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Vera M
VM
Morsch
Laboratory of Experimental Surgery, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Diego V
DV
Beckmann
Departament of Microbiology and Parasitology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Lívia G
LG
Castilhos
Departament of Microbiology and Parasitology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Maria L P
MLP
Thorstenberg
Departament of Microbiology and Parasitology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Jeandre A Dos S
JADS
Jaques
Departament of Microbiology and Parasitology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Viviane do C G
VDCG
Souza
Departament of Biochemistry and Molecular Biology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Júlia G
JG
Farias
Departament of Biochemistry and Molecular Biology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Caroline C
CC
Martins
Departament of Biochemistry and Molecular Biology, Federal University of Santa Maria, Av. Roraima, 97105-900 Santa Maria, RS, Brazil.
Maria R C
MRC
Schetinger
D000227:Adenine Nucleotides / Q000378:metabolism*; D000243:Adenosine Deaminase / Q000378:metabolism; D000818:Animals; D001792:Blood Platelets / Q000187:drug effects / Q000201:enzymology*; D002726:Chlorogenic Acid / Q000737:chemistry / Q000494:pharmacology*; D003711:Demyelinating Diseases / Q000097:blood* / Q000139:chemically induced* / Q000201:enzymology; D004996:Ethidium; D006868:Hydrolysis; D008297:Male; D010974:Platelet Aggregation / Q000187:drug effects; D016133:Polymerase Chain Reaction; D011755:Pyrophosphatases / Q000378:metabolism; D017208:Rats, Wistar
D000227:Adenine Nucleotides; D002726:Chlorogenic Acid; D000243:Adenosine Deaminase; D011755:Pyrophosphatases; C021899:nucleotide pyrophosphatase; D004996:Ethidium
Adenosine deaminase; Chlorogenic acid; Demyelination; Ectonucleotidase; Platelets