Effects of platelets on the protein expression in aortic segments: A proteomic approach.
It is well known the effects of the vascular wall on platelet activity but little is known about the effects of platelets on the proteins expression in the vascular wall. We analyzed whether platelets may modify the protein expression in the vascular wall. We used an in vitro model coincubating human platelet rich plasma (PRP) with control and 10 ng/ml tumor necrosis factor-α (TNF-α)-preincubated bovine aortic segments. 2DE, mass spectrometry and Western blot analysis were used to determine changes in the expression of proteins associated with the cytoskeleton and energetic metabolism in the aortic segments. In control healthy vascular wall, only the cytoskeleton-related proteins expression was modified by PRP. However, when PRP was coincubated with TNF-α pre-stimulated aortic segments lesser number of cytoskeleton-related proteins were modified. With respect to energetic metabolism, in control segments, PRP failed to modify any of the analyzed energetic-related proteins. However, in TNF-α-preincubated segments the presence of PRP upexpressed glyceraldehyde-3-phosphate dehydrogenase. Moreover, by western blot experiments it was observed that in TNF-α-preincubated segments the expression of fructose 1,6-bisphosphate aldolase was downregulated by platelets. However, no differences were found in the expression of triosephosphate isomerase and ATP synthase α-chain. In addition, the activity of fructose 1,6-bisphosphate aldolase and piruvate content was significantly reduced without modification on triosephosphate isomerase activity. In conclusion, the crosstalk between platelets and vascular wall is bidirectional and platelets regulated in the vascular wall the expression of proteins associated with the cytoskeleton and energetic metabolism, particularly in the healthy vascular wall.
Modrego
Javier
J
Cardiovascular Research Unit, Cardiology Service, Cardiovascular Institute, Hospital Clínico San Carlos, Madrid, Spain.
Moñux
Guillermo
G
Mateos-Cáceres
Petra J
PJ
Martínez-López
Isaac
I
Segura
Antonio
A
Zamorano-León
José J
JJ
Rodríguez-Sierra
Pablo
P
Serrano
Javier
J
Macaya
Carlos
C
López-Farré
Antonio J
AJ
D000328:Adult; D000595:Amino Acid Sequence; D000818:Animals; D001011:Aorta / Q000378:metabolism*; D001792:Blood Platelets / Q000378:metabolism*; D015153:Blotting, Western; D002417:Cattle; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004734:Energy Metabolism; D006801:Humans; D066298:In Vitro Techniques; D008969:Molecular Sequence Data; D053657:Platelet-Rich Plasma / Q000378:metabolism; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*
D003598:Cytoskeletal Proteins; D011506:Proteins