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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

Inhibitory effects of CA on granule secretion, P-selectin expression, ATP release, and Ca2+ mobilization in ADP stimulated platelets.Platelets were preincubated with or without CA in the presence of CaCl2 (1 mM) at 37 °C for 20 min. The cells were then incubated with ADP (20 μM) for 20 min before P-selectin expression (A,B), ATP release (C), and Ca2+ levels (D) were analyzed. (A) A typical flow cytometry result showing P-selectin expression in different conditions. (B) A quantitative assessment of P-selectin expression in different conditions with data from at least four independent experiments. (C) A quantitative assessment of ATP release in different conditions. (D) A quantitative assessment of intracellular calcium mobilization in different conditions. Data are expressed as means ± SEM (n = 10). *p < 0.05 vs control group, #p < 0.05 vs ADP group.
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f5: Inhibitory effects of CA on granule secretion, P-selectin expression, ATP release, and Ca2+ mobilization in ADP stimulated platelets.Platelets were preincubated with or without CA in the presence of CaCl2 (1 mM) at 37 °C for 20 min. The cells were then incubated with ADP (20 μM) for 20 min before P-selectin expression (A,B), ATP release (C), and Ca2+ levels (D) were analyzed. (A) A typical flow cytometry result showing P-selectin expression in different conditions. (B) A quantitative assessment of P-selectin expression in different conditions with data from at least four independent experiments. (C) A quantitative assessment of ATP release in different conditions. (D) A quantitative assessment of intracellular calcium mobilization in different conditions. Data are expressed as means ± SEM (n = 10). *p < 0.05 vs control group, #p < 0.05 vs ADP group.

Mentions: Since granule secretions are critical marker of platelet activation prior to aggregation, therefore, the effect of CA on ADP-activated granule secretion was examined in the present study. As shown in Fig. 5A,B, stimulation with ADP (20 μM) significantly enhanced P-selectin surface expression in platelets, as compared to that in control untreated platelets. Interestingly, this increase in P-selectin expression was significantly reduced by CA (5–100 μM) in a dose-dependent manner. A similar inhibitory effect of CA was observed for ATP release from platelets (Fig. 5C). Taking together, these results indicate that CA pretreatment inhibits secretory activity of both platelet α-granule and dense granule.


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)

Inhibitory effects of CA on granule secretion, P-selectin expression, ATP release, and Ca2+ mobilization in ADP stimulated platelets.Platelets were preincubated with or without CA in the presence of CaCl2 (1 mM) at 37 °C for 20 min. The cells were then incubated with ADP (20 μM) for 20 min before P-selectin expression (A,B), ATP release (C), and Ca2+ levels (D) were analyzed. (A) A typical flow cytometry result showing P-selectin expression in different conditions. (B) A quantitative assessment of P-selectin expression in different conditions with data from at least four independent experiments. (C) A quantitative assessment of ATP release in different conditions. (D) A quantitative assessment of intracellular calcium mobilization in different conditions. Data are expressed as means ± SEM (n = 10). *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

f5: Inhibitory effects of CA on granule secretion, P-selectin expression, ATP release, and Ca2+ mobilization in ADP stimulated platelets.Platelets were preincubated with or without CA in the presence of CaCl2 (1 mM) at 37 °C for 20 min. The cells were then incubated with ADP (20 μM) for 20 min before P-selectin expression (A,B), ATP release (C), and Ca2+ levels (D) were analyzed. (A) A typical flow cytometry result showing P-selectin expression in different conditions. (B) A quantitative assessment of P-selectin expression in different conditions with data from at least four independent experiments. (C) A quantitative assessment of ATP release in different conditions. (D) A quantitative assessment of intracellular calcium mobilization in different conditions. Data are expressed as means ± SEM (n = 10). *p < 0.05 vs control group, #p < 0.05 vs ADP group.
Mentions: Since granule secretions are critical marker of platelet activation prior to aggregation, therefore, the effect of CA on ADP-activated granule secretion was examined in the present study. As shown in Fig. 5A,B, stimulation with ADP (20 μM) significantly enhanced P-selectin surface expression in platelets, as compared to that in control untreated platelets. Interestingly, this increase in P-selectin expression was significantly reduced by CA (5–100 μM) in a dose-dependent manner. A similar inhibitory effect of CA was observed for ATP release from platelets (Fig. 5C). Taking together, these results indicate that CA pretreatment inhibits secretory activity of both platelet α-granule and dense granule.

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