S V Kansra | PubMed Health

PubMed2000

Reverse relationship between mitogen activated protein kinase and human platelet aggregation.

The role of MAPK in platelets was investigated. In human platelets maintained at 4 degrees C for 2 hr, the MAPK activity increased (approximately 2 fold) when compared to those maintained at 37 degrees C. When aggregation was monitored under these conditions, the platelets maintained at 4 degrees C or 15 degrees C showed an 85% and 71% decrease respectively to thrombin (0.5 U/ml for 1 min) induced aggregation. When the platelet cytosol was maintained at 4 degrees C and assayed for MAPK activity, the MAPK activity decreased significantly, indicating that the observed effects are seen in intact platelets only, and are not due to temperature effects on the assay. When platelets maintained at 4 degrees C or 15 degrees C (for 2 hrs) were transferred to 37 degrees C, the MAPK activity decreased to levels observed in platelets maintained throughout at 37 degrees C and was thus reversible. Therefore, it is concluded that a possible reverse relationship between MAPK and platelet aggregation plays a role in platelet responses.

Blood Platelets↗

PubMed1999

Activation of mitogen activated protein kinase in human platelets by genistein.

Genistein, a putative tyrosine kinase inhibitor, stimulated human platelet mitogen activated protein kinase (MAPK) activity in a dose- and time-dependent manner. When MAPK was maximally stimulated by phorbol 12-myristate 13-acetate (PMA), genistein still elicited the increase in MAPK activity. Staurosporine (50 nm), significantly decreased the PMA-induced MAPK activity, but had little inhibitory effect on the genistein-induced MAPK activity. Both these observations indicated a protein kinase C (PKC) independent pathway for the genistein-stimulated MAPK activity. When other tyrosine kinase inhibitors (methyl-2,5-dihydroxycinnamate, and tyrphostin) were employed, similar increases in the MAPK activity were observed. Addition of genistein to cytosolic fraction of platelets had no effect on the MAPK activity and indicated that this effect is not due to direct physical interaction between genistein and MAPK and that intact platelets are required for it. MAPK activity of platelets from rabbit and pig was also stimulated by genistein. This effect of genistein was not observed in other cell types tested (BNLCL2, HEL and U937 cells). Forskolin, which increases cyclic AMP had little effect on the basal platelet MAPK activity or the genistein activated MAPK, while it decreased by half the PMA-induced MAPK activity. The inactive analog of genistein, daidzein, which does not inhibit tyrosine kinase had little effect on MAPK. Genistein caused a decrease in basal tyrosine phosphorylation of pp60(c-src) protein as detected with anti-phosphotyrosine (anti-PTyr) Ab. Thus, inhibition of basal tyrosine kinase results in an increase in MAPK activity. This study demonstrates for the first time a novel mechanism for regulation of MAPK in platelets in which inhibition of tyrosine kinase results in activation of MAPK, independent of PKC and cAMP pathways.

Animals↗