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Inhibitory effect of caffeic acid on ADP-induced thrombus formation and platelet activation involves mitogen-activated protein kinases.

Lu Y, Li Q, Liu YY, Sun K, Fan JY, Wang CS, Han JY - Sci Rep (2015)

Bottom Line: In this study, we investigated the antithrombotic effect of CA in mouse cerebral arterioles and venules using intravital microscopy.Our results demonstrated that CA (1.25-5 mg/kg) significantly inhibited thrombus formation in vivo.Additionally, CA attenuated p38, ERK, and JNK activation, and enhanced cAMP levels.

View Article: PubMed Central - PubMed

Affiliation: Department of gynaecology, Beijing Royal Integrative Medicine Hospital, Beijing, China.

ABSTRACT
Caffeic acid (CA), one of the active constituents of Radix Salvia miltiorrhizae, exhibits antioxidant and anti-inflammatory activities. However, few studies have assessed the ability of CA to inhibit platelet mediated thrombus generation in vivo. In this study, we investigated the antithrombotic effect of CA in mouse cerebral arterioles and venules using intravital microscopy. The antiplatelet activity of CA in ADP stimulated mouse platelets in vitro was also examined in attempt to explore the underlying mechanism. Our results demonstrated that CA (1.25-5 mg/kg) significantly inhibited thrombus formation in vivo. In vitro, CA (25-100 μM) inhibited ADP-induced platelet aggregation, P-selectin expression, ATP release, Ca(2+) mobilization, and integrin αIIbβ3 activation. Additionally, CA attenuated p38, ERK, and JNK activation, and enhanced cAMP levels. Taken together, these data provide evidence for the inhibition of CA on platelet-mediated thrombosis in vivo, which is, at least partly, mediated by interference in phosphorylation of ERK, p38, and JNK leading to elevation of cAMP and down-regulation of P-selectin expression and αIIbβ3 activation. These results suggest that CA may have potential for the treatment of aberrant platelet activation-related diseases.

No MeSH data available.


Related in: MedlinePlus

CA attenuates ADP-activated platelet p38MAPK, ERK and JNK phosphorylation.Washed platelets were incubated with CA or vehicle for 20 min and then stimulated with ADP for 20 min. Platelet proteins were then extracted and specific antibodies were used to measure the levels of total and phosphorylated p38 (A), ERK (B), JNK (C), and Akt (D). CA attenuated the phosphorylation of p38, ERK and JNK but not of Akt (Ser473) in the activated platelets. Images are representative of three separate experiments. The relative protein expression levels were quantified by Quantity One software. Data represent the mean ± SEM of at least three independent experiments performed in triplicate. *p < 0.05 vs control group; #p < 0.05 vs ADP group.
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f9: CA attenuates ADP-activated platelet p38MAPK, ERK and JNK phosphorylation.Washed platelets were incubated with CA or vehicle for 20 min and then stimulated with ADP for 20 min. Platelet proteins were then extracted and specific antibodies were used to measure the levels of total and phosphorylated p38 (A), ERK (B), JNK (C), and Akt (D). CA attenuated the phosphorylation of p38, ERK and JNK but not of Akt (Ser473) in the activated platelets. Images are representative of three separate experiments. The relative protein expression levels were quantified by Quantity One software. Data represent the mean ± SEM of at least three independent experiments performed in triplicate. *p < 0.05 vs control group; #p < 0.05 vs ADP group.

Mentions: Phosphorylation of MAPKs, including ERK, p38, and JNK, and Akt in platelets are closely associated with platelet activation and aggregation. Hence, we determined whether CA inhibited MAPKs and Akt phosphorylations in ADP stimulated platelets. The immunoblotting analysis revealed that treatment with ADP (20 μM) provoked marked p38, ERK, JNK and Akt phosphorylation in mouse platelets. Pretreatment with CA dose-dependently suppressed ADP-induced p38 MAPK (Fig. 9A), ERK (Fig. 9B) and JNK (Fig. 9C) phosphorylation. CA did not affect phosphorylation of Akt (Ser473) (Fig. 9D), but significantly reduced phosphorylation of P-Akt (Thr 308) (Supplemental Fig. 4). These data suggest that the MAPKs (ERK, p38, and JNK) and Akt signaling pathways are implicated in the effects of CA.


Inhibitory effect of caffeic acid on ADP-induced thrombus formation and platelet activation involves mitogen-activated protein kinases.

Lu Y, Li Q, Liu YY, Sun K, Fan JY, Wang CS, Han JY - Sci Rep (2015)

CA attenuates ADP-activated platelet p38MAPK, ERK and JNK phosphorylation.Washed platelets were incubated with CA or vehicle for 20 min and then stimulated with ADP for 20 min. Platelet proteins were then extracted and specific antibodies were used to measure the levels of total and phosphorylated p38 (A), ERK (B), JNK (C), and Akt (D). CA attenuated the phosphorylation of p38, ERK and JNK but not of Akt (Ser473) in the activated platelets. Images are representative of three separate experiments. The relative protein expression levels were quantified by Quantity One software. Data represent the mean ± SEM of at least three independent experiments performed in triplicate. *p < 0.05 vs control group; #p < 0.05 vs ADP group.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4561902&req=5

f9: CA attenuates ADP-activated platelet p38MAPK, ERK and JNK phosphorylation.Washed platelets were incubated with CA or vehicle for 20 min and then stimulated with ADP for 20 min. Platelet proteins were then extracted and specific antibodies were used to measure the levels of total and phosphorylated p38 (A), ERK (B), JNK (C), and Akt (D). CA attenuated the phosphorylation of p38, ERK and JNK but not of Akt (Ser473) in the activated platelets. Images are representative of three separate experiments. The relative protein expression levels were quantified by Quantity One software. Data represent the mean ± SEM of at least three independent experiments performed in triplicate. *p < 0.05 vs control group; #p < 0.05 vs ADP group.
Mentions: Phosphorylation of MAPKs, including ERK, p38, and JNK, and Akt in platelets are closely associated with platelet activation and aggregation. Hence, we determined whether CA inhibited MAPKs and Akt phosphorylations in ADP stimulated platelets. The immunoblotting analysis revealed that treatment with ADP (20 μM) provoked marked p38, ERK, JNK and Akt phosphorylation in mouse platelets. Pretreatment with CA dose-dependently suppressed ADP-induced p38 MAPK (Fig. 9A), ERK (Fig. 9B) and JNK (Fig. 9C) phosphorylation. CA did not affect phosphorylation of Akt (Ser473) (Fig. 9D), but significantly reduced phosphorylation of P-Akt (Thr 308) (Supplemental Fig. 4). These data suggest that the MAPKs (ERK, p38, and JNK) and Akt signaling pathways are implicated in the effects of CA.

Bottom Line: In this study, we investigated the antithrombotic effect of CA in mouse cerebral arterioles and venules using intravital microscopy.Our results demonstrated that CA (1.25-5 mg/kg) significantly inhibited thrombus formation in vivo.Additionally, CA attenuated p38, ERK, and JNK activation, and enhanced cAMP levels.

View Article: PubMed Central - PubMed

Affiliation: Department of gynaecology, Beijing Royal Integrative Medicine Hospital, Beijing, China.

ABSTRACT
Caffeic acid (CA), one of the active constituents of Radix Salvia miltiorrhizae, exhibits antioxidant and anti-inflammatory activities. However, few studies have assessed the ability of CA to inhibit platelet mediated thrombus generation in vivo. In this study, we investigated the antithrombotic effect of CA in mouse cerebral arterioles and venules using intravital microscopy. The antiplatelet activity of CA in ADP stimulated mouse platelets in vitro was also examined in attempt to explore the underlying mechanism. Our results demonstrated that CA (1.25-5 mg/kg) significantly inhibited thrombus formation in vivo. In vitro, CA (25-100 μM) inhibited ADP-induced platelet aggregation, P-selectin expression, ATP release, Ca(2+) mobilization, and integrin αIIbβ3 activation. Additionally, CA attenuated p38, ERK, and JNK activation, and enhanced cAMP levels. Taken together, these data provide evidence for the inhibition of CA on platelet-mediated thrombosis in vivo, which is, at least partly, mediated by interference in phosphorylation of ERK, p38, and JNK leading to elevation of cAMP and down-regulation of P-selectin expression and αIIbβ3 activation. These results suggest that CA may have potential for the treatment of aberrant platelet activation-related diseases.

No MeSH data available.


Related in: MedlinePlus