Limits...
Effect of Cordycepin-Enriched WIB801C from Cordyceps militaris Suppressing Fibrinogen Binding to Glycoprotein IIb/IIIa.

Lee DH, Kim HH, Lim DH, Kim JL, Park HJ - Biomol Ther (Seoul) (2015)

Bottom Line: CE-WIB801C and cordycepin stimulated the phosphorylation of VASP (Ser(157)) and the dephosphorylation of PI3K and Akt, and inhibited the binding of fibrinogen to glycoprotein IIb/IIIa (αIIb/β3) and the release of ATP and serotonin in collagen-induced platelet aggregation.A-kinase inhibitor Rp-8-Br-cAMPS reduced CE-WIB801C-, and cordycepin-increased VASP (Ser(157)) phosphorylation, and increased CE-WIB801C-, and cordycepin-inhibited the fibrinogen binding to αIIb/β3.Therefore, we demonstrate that CE-WIB801C-, and cordycepin-inhibited fibrinogen binding to αIIb/β3 are due to stimulation of cAMP-dependent phosphorylation of VASP (Ser(157)), and inhibition of PI3K/Akt phosphorylation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749.

ABSTRACT
In this study, we investigated the effects of cordycepin-enriched (CE)-WIB801C, a n-butanol extract of Cordyceps militaris-hypha on collagen-stimulated platelet aggregation. CE-WIB801C dose dependently inhibited collagen-induced platelet aggregation, and had a synergistic effect together with cordycepin (W-cordycepin) from CE-WIB801C on the inhibition of collagen-induced platelet aggregation. CE-WIB801C and cordycepin stimulated the phosphorylation of VASP (Ser(157)) and the dephosphorylation of PI3K and Akt, and inhibited the binding of fibrinogen to glycoprotein IIb/IIIa (αIIb/β3) and the release of ATP and serotonin in collagen-induced platelet aggregation. A-kinase inhibitor Rp-8-Br-cAMPS reduced CE-WIB801C-, and cordycepin-increased VASP (Ser(157)) phosphorylation, and increased CE-WIB801C-, and cordycepin-inhibited the fibrinogen binding to αIIb/β3. Therefore, we demonstrate that CE-WIB801C-, and cordycepin-inhibited fibrinogen binding to αIIb/β3 are due to stimulation of cAMP-dependent phosphorylation of VASP (Ser(157)), and inhibition of PI3K/Akt phosphorylation. These results strongly indicate that CE-WIB801C and cordycepin may have preventive or therapeutic potential for platelet aggregation-mediated diseases, such as thrombosis, myocardial infarction, atherosclerosis, and ischemic cerebrovascular disease.

No MeSH data available.


Related in: MedlinePlus

Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding in the presence of A-kinase inhibitor (Rp-8-BrcAMPS), or G-kinase inhibitor (Rp-8-Br-cGMPS). (A) The flow cytometry histograms on fibrinogen binding. a, Collagen (10 μg/mL)+CEWIB801C (400 μg/mL)+Rp-8-Br-cAMPS (250 μM); b, Collagen (10 μg/mL)+CE-WIB801C (400 μg/mL)+Rp-8-Br-cGMPS (250 μM); c, Collagen (10 μg/mL)+cordycepin (500 μM)+Rp-8-Br-cAMPS (250 μM); d, Collagen (10 μg/mL)+cordycepin (500 μM)+ Rp-8-Br-cGMPS (250 μM); e, Collagen (10 μg/mL)+pCPT-cAMP (1 mM); f, Collagen (10 μg/mL)+8-Br-cGMP (1 mM). (B) Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding (%) in the presence of A-kinase inhibitor (Rp-8-Br-cAMPS) or G-kinase inhibitor (Rp-8-Br-cGMPS). Determination of fibrinogen binding to αIIb/β3 was carried out as described in “Materials and Methods.” The data are expressed as the mean ± S.E.M. (n=4). **p<0.001 versus the collagen-stimulated platelets, ††p<0.001 versus the collagen-stimulated platelets in the presence of CEWIB801C (400 μg/mL). #p<0.05 versus the collagen-stimulated platelets in the presence of cordycepin (500 μM). ##p<0.001 versus the collagen- stimulated platelets in the presence of cordycepin (500 μM).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4286751&req=5

f5-bt-23-60: Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding in the presence of A-kinase inhibitor (Rp-8-BrcAMPS), or G-kinase inhibitor (Rp-8-Br-cGMPS). (A) The flow cytometry histograms on fibrinogen binding. a, Collagen (10 μg/mL)+CEWIB801C (400 μg/mL)+Rp-8-Br-cAMPS (250 μM); b, Collagen (10 μg/mL)+CE-WIB801C (400 μg/mL)+Rp-8-Br-cGMPS (250 μM); c, Collagen (10 μg/mL)+cordycepin (500 μM)+Rp-8-Br-cAMPS (250 μM); d, Collagen (10 μg/mL)+cordycepin (500 μM)+ Rp-8-Br-cGMPS (250 μM); e, Collagen (10 μg/mL)+pCPT-cAMP (1 mM); f, Collagen (10 μg/mL)+8-Br-cGMP (1 mM). (B) Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding (%) in the presence of A-kinase inhibitor (Rp-8-Br-cAMPS) or G-kinase inhibitor (Rp-8-Br-cGMPS). Determination of fibrinogen binding to αIIb/β3 was carried out as described in “Materials and Methods.” The data are expressed as the mean ± S.E.M. (n=4). **p<0.001 versus the collagen-stimulated platelets, ††p<0.001 versus the collagen-stimulated platelets in the presence of CEWIB801C (400 μg/mL). #p<0.05 versus the collagen-stimulated platelets in the presence of cordycepin (500 μM). ##p<0.001 versus the collagen- stimulated platelets in the presence of cordycepin (500 μM).

Mentions: Next, we investigated whether the VASP phosphorylation by CE-WIB801C involved in inhibition of fibrinogen binding to αIIb/β3. As shown in Fig. 4, collagen activated fibrinogen binding to αIIb/β3 (Fig. 4A–b), and increased the degree of fibrinogen binding to αIIb/β3 up to 77.2 ± 5.5% as compared with that (5.4 ± 0.2%) of intact platelets, basal (Table 1). However, CE-WIB801C inhibited collagen-activated fibrinogen binding to αIIb/β3 (Fig. 4A–c, 4B), and its inhibitory degree was 89.2% as compared with that (77.2 ± 5.5%) by collagen (Table 1). Cordycepin also potently inhibited collagen-activated fibrinogen binding to αIIb/β3 (Fig. 4A–d and 4B). Because the inhibition of αIIb/β3 is resulted from cAMP/A-kinase- and cGMP/G-kinase- mediated VASP phosphorylation, and it is known that cAMP- and cGMP-increasing compounds involve in inhibition of αIIb/β3 (Horstrup et al., 1994; Barragan et al., 2003), we investigated whether the inhibition of fibrinogen binding to αIIb/β3 by CE-WIB801C was contributed to which kinase of Akinase and G-kinase. A-kinase activator pCPT-cAMP and G-kinase activator 8-Br-cGMP inhibited collagen-induced fibrinogen binding to αIIb/β3, respectively (Fig. 5A–e, f and Fig. 5B). It was confirmed that cAMP/A-kinase and cGMP/G-kinase pathway involve in inhibition of fibrinogen binding to αIIb/β3 in collagen-induced platelet aggregation. CE-WIB801C-inhibited fibrinogen binding to αIIb/β3 (Fig. 4A–c and Fig. 4B) was elevated by A-kinase inhibitor Rp-8-Br-cAMPS (Fig. 5A–a and Fig. 5B), and its stimulatory degree was 386.7% as compared with that (8.3%) by both CE-WIB801C and collagen (Table 1). However, CE-WIB801C-inhibited fibrinogen binding to αIIb/β3 (Fig. 4A–c and Fig. 4B) was not almost elevated by G-kinase inhibitor Rp-8-Br-cGMPS (Fig. 5A–b and Fig. 5B), and its stimulatory degree was 6.0% as compared with that (8.3%) by both CE-WIB801C and collagen (Table 1).


Effect of Cordycepin-Enriched WIB801C from Cordyceps militaris Suppressing Fibrinogen Binding to Glycoprotein IIb/IIIa.

Lee DH, Kim HH, Lim DH, Kim JL, Park HJ - Biomol Ther (Seoul) (2015)

Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding in the presence of A-kinase inhibitor (Rp-8-BrcAMPS), or G-kinase inhibitor (Rp-8-Br-cGMPS). (A) The flow cytometry histograms on fibrinogen binding. a, Collagen (10 μg/mL)+CEWIB801C (400 μg/mL)+Rp-8-Br-cAMPS (250 μM); b, Collagen (10 μg/mL)+CE-WIB801C (400 μg/mL)+Rp-8-Br-cGMPS (250 μM); c, Collagen (10 μg/mL)+cordycepin (500 μM)+Rp-8-Br-cAMPS (250 μM); d, Collagen (10 μg/mL)+cordycepin (500 μM)+ Rp-8-Br-cGMPS (250 μM); e, Collagen (10 μg/mL)+pCPT-cAMP (1 mM); f, Collagen (10 μg/mL)+8-Br-cGMP (1 mM). (B) Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding (%) in the presence of A-kinase inhibitor (Rp-8-Br-cAMPS) or G-kinase inhibitor (Rp-8-Br-cGMPS). Determination of fibrinogen binding to αIIb/β3 was carried out as described in “Materials and Methods.” The data are expressed as the mean ± S.E.M. (n=4). **p<0.001 versus the collagen-stimulated platelets, ††p<0.001 versus the collagen-stimulated platelets in the presence of CEWIB801C (400 μg/mL). #p<0.05 versus the collagen-stimulated platelets in the presence of cordycepin (500 μM). ##p<0.001 versus the collagen- stimulated platelets in the presence of cordycepin (500 μM).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4286751&req=5

f5-bt-23-60: Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding in the presence of A-kinase inhibitor (Rp-8-BrcAMPS), or G-kinase inhibitor (Rp-8-Br-cGMPS). (A) The flow cytometry histograms on fibrinogen binding. a, Collagen (10 μg/mL)+CEWIB801C (400 μg/mL)+Rp-8-Br-cAMPS (250 μM); b, Collagen (10 μg/mL)+CE-WIB801C (400 μg/mL)+Rp-8-Br-cGMPS (250 μM); c, Collagen (10 μg/mL)+cordycepin (500 μM)+Rp-8-Br-cAMPS (250 μM); d, Collagen (10 μg/mL)+cordycepin (500 μM)+ Rp-8-Br-cGMPS (250 μM); e, Collagen (10 μg/mL)+pCPT-cAMP (1 mM); f, Collagen (10 μg/mL)+8-Br-cGMP (1 mM). (B) Effects of CE-WIB801C and cordycepin on collagen-induced fibrinogen binding (%) in the presence of A-kinase inhibitor (Rp-8-Br-cAMPS) or G-kinase inhibitor (Rp-8-Br-cGMPS). Determination of fibrinogen binding to αIIb/β3 was carried out as described in “Materials and Methods.” The data are expressed as the mean ± S.E.M. (n=4). **p<0.001 versus the collagen-stimulated platelets, ††p<0.001 versus the collagen-stimulated platelets in the presence of CEWIB801C (400 μg/mL). #p<0.05 versus the collagen-stimulated platelets in the presence of cordycepin (500 μM). ##p<0.001 versus the collagen- stimulated platelets in the presence of cordycepin (500 μM).
Mentions: Next, we investigated whether the VASP phosphorylation by CE-WIB801C involved in inhibition of fibrinogen binding to αIIb/β3. As shown in Fig. 4, collagen activated fibrinogen binding to αIIb/β3 (Fig. 4A–b), and increased the degree of fibrinogen binding to αIIb/β3 up to 77.2 ± 5.5% as compared with that (5.4 ± 0.2%) of intact platelets, basal (Table 1). However, CE-WIB801C inhibited collagen-activated fibrinogen binding to αIIb/β3 (Fig. 4A–c, 4B), and its inhibitory degree was 89.2% as compared with that (77.2 ± 5.5%) by collagen (Table 1). Cordycepin also potently inhibited collagen-activated fibrinogen binding to αIIb/β3 (Fig. 4A–d and 4B). Because the inhibition of αIIb/β3 is resulted from cAMP/A-kinase- and cGMP/G-kinase- mediated VASP phosphorylation, and it is known that cAMP- and cGMP-increasing compounds involve in inhibition of αIIb/β3 (Horstrup et al., 1994; Barragan et al., 2003), we investigated whether the inhibition of fibrinogen binding to αIIb/β3 by CE-WIB801C was contributed to which kinase of Akinase and G-kinase. A-kinase activator pCPT-cAMP and G-kinase activator 8-Br-cGMP inhibited collagen-induced fibrinogen binding to αIIb/β3, respectively (Fig. 5A–e, f and Fig. 5B). It was confirmed that cAMP/A-kinase and cGMP/G-kinase pathway involve in inhibition of fibrinogen binding to αIIb/β3 in collagen-induced platelet aggregation. CE-WIB801C-inhibited fibrinogen binding to αIIb/β3 (Fig. 4A–c and Fig. 4B) was elevated by A-kinase inhibitor Rp-8-Br-cAMPS (Fig. 5A–a and Fig. 5B), and its stimulatory degree was 386.7% as compared with that (8.3%) by both CE-WIB801C and collagen (Table 1). However, CE-WIB801C-inhibited fibrinogen binding to αIIb/β3 (Fig. 4A–c and Fig. 4B) was not almost elevated by G-kinase inhibitor Rp-8-Br-cGMPS (Fig. 5A–b and Fig. 5B), and its stimulatory degree was 6.0% as compared with that (8.3%) by both CE-WIB801C and collagen (Table 1).

Bottom Line: CE-WIB801C and cordycepin stimulated the phosphorylation of VASP (Ser(157)) and the dephosphorylation of PI3K and Akt, and inhibited the binding of fibrinogen to glycoprotein IIb/IIIa (αIIb/β3) and the release of ATP and serotonin in collagen-induced platelet aggregation.A-kinase inhibitor Rp-8-Br-cAMPS reduced CE-WIB801C-, and cordycepin-increased VASP (Ser(157)) phosphorylation, and increased CE-WIB801C-, and cordycepin-inhibited the fibrinogen binding to αIIb/β3.Therefore, we demonstrate that CE-WIB801C-, and cordycepin-inhibited fibrinogen binding to αIIb/β3 are due to stimulation of cAMP-dependent phosphorylation of VASP (Ser(157)), and inhibition of PI3K/Akt phosphorylation.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical Laboratory Science, College of Biomedical Science and Engineering, Inje University, Gimhae 621-749.

ABSTRACT
In this study, we investigated the effects of cordycepin-enriched (CE)-WIB801C, a n-butanol extract of Cordyceps militaris-hypha on collagen-stimulated platelet aggregation. CE-WIB801C dose dependently inhibited collagen-induced platelet aggregation, and had a synergistic effect together with cordycepin (W-cordycepin) from CE-WIB801C on the inhibition of collagen-induced platelet aggregation. CE-WIB801C and cordycepin stimulated the phosphorylation of VASP (Ser(157)) and the dephosphorylation of PI3K and Akt, and inhibited the binding of fibrinogen to glycoprotein IIb/IIIa (αIIb/β3) and the release of ATP and serotonin in collagen-induced platelet aggregation. A-kinase inhibitor Rp-8-Br-cAMPS reduced CE-WIB801C-, and cordycepin-increased VASP (Ser(157)) phosphorylation, and increased CE-WIB801C-, and cordycepin-inhibited the fibrinogen binding to αIIb/β3. Therefore, we demonstrate that CE-WIB801C-, and cordycepin-inhibited fibrinogen binding to αIIb/β3 are due to stimulation of cAMP-dependent phosphorylation of VASP (Ser(157)), and inhibition of PI3K/Akt phosphorylation. These results strongly indicate that CE-WIB801C and cordycepin may have preventive or therapeutic potential for platelet aggregation-mediated diseases, such as thrombosis, myocardial infarction, atherosclerosis, and ischemic cerebrovascular disease.

No MeSH data available.


Related in: MedlinePlus