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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

The inhibitory effect of CA on thrombosis in mouse cerebral venule.(A) Presented are the representative images of thrombus formation in mouse cerebral venules in control, ADP, and CA + ADP group, at baseline (1), 5 (2), 15 (3), 25 (4), 40 (5), 50 (6), and 60 (7) min, respectively. Selected venules (V) have a diameter of 35–45 μm. Arrows indicate thrombus (T) formed in the venular lumen. (B) The effect of CA on time course of the area ratio of thrombus to mouse cerebral venule. (C) The effect of CA on accumulative area ratio of thrombus to mouse cerebral venule during 60 min. ADP: the animals were treated with ADP at 0, 5, 15, and 25 min, respectively. CA + ADP: the animals were treated with CA (5 mg/kg) starting from 20 min before thrombus induction. Data are expressed as means ± SEM (n = 10). *p < 0.05 vs control group, #p < 0.05 vs ADP group.
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f3: The inhibitory effect of CA on thrombosis in mouse cerebral venule.(A) Presented are the representative images of thrombus formation in mouse cerebral venules in control, ADP, and CA + ADP group, at baseline (1), 5 (2), 15 (3), 25 (4), 40 (5), 50 (6), and 60 (7) min, respectively. Selected venules (V) have a diameter of 35–45 μm. Arrows indicate thrombus (T) formed in the venular lumen. (B) The effect of CA on time course of the area ratio of thrombus to mouse cerebral venule. (C) The effect of CA on accumulative area ratio of thrombus to mouse cerebral venule during 60 min. ADP: the animals were treated with ADP at 0, 5, 15, and 25 min, respectively. CA + ADP: the animals were treated with CA (5 mg/kg) starting from 20 min before thrombus induction. Data are expressed as means ± SEM (n = 10). *p < 0.05 vs control group, #p < 0.05 vs ADP group.

Mentions: We next assessed the effect of CA on thrombus formation in mouse cerebral venules induced by ADP. As shown in Fig. 3A, only few of fluorescently labeled platelets (bright dots) were detected transiently adhered on the cerebral venular wall in all groups before the topical application of ADP (a1, b1, and c1), which persisted in the control group over the entire observation (a2–a7). In contrast, in ADP group, platelet adhesion was rapidly enhanced on the venular wall in mice, followed by a gradual build-up of platelet thrombus, in which fluorescently labeled platelets were seen as bright dots. The thrombus grew rapidly and obstructed partially the lumen of the vessel in a short time, developing to a comparable size to the venular diameter until it flowed downstream (b2–b4). The blood flow decreased as the thrombus grew, falling quickly over the last minutes and forming a stable thrombus (b5–b7). In the mice pretreated with CA (5 mg/kg), topical application of ADP-induced platelet adhesion and thrombus formation were markedly attenuated, as shown by the delayed thrombus formation and fewer accumulated platelets (c2–c4). After stopping ADP stimulation, thrombus disrupted into non-uniform fragments, which were quickly carried away by the blood flow (c5–c57).


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)

The inhibitory effect of CA on thrombosis in mouse cerebral venule.(A) Presented are the representative images of thrombus formation in mouse cerebral venules in control, ADP, and CA + ADP group, at baseline (1), 5 (2), 15 (3), 25 (4), 40 (5), 50 (6), and 60 (7) min, respectively. Selected venules (V) have a diameter of 35–45 μm. Arrows indicate thrombus (T) formed in the venular lumen. (B) The effect of CA on time course of the area ratio of thrombus to mouse cerebral venule. (C) The effect of CA on accumulative area ratio of thrombus to mouse cerebral venule during 60 min. ADP: the animals were treated with ADP at 0, 5, 15, and 25 min, respectively. CA + ADP: the animals were treated with CA (5 mg/kg) starting from 20 min before thrombus induction. 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

f3: The inhibitory effect of CA on thrombosis in mouse cerebral venule.(A) Presented are the representative images of thrombus formation in mouse cerebral venules in control, ADP, and CA + ADP group, at baseline (1), 5 (2), 15 (3), 25 (4), 40 (5), 50 (6), and 60 (7) min, respectively. Selected venules (V) have a diameter of 35–45 μm. Arrows indicate thrombus (T) formed in the venular lumen. (B) The effect of CA on time course of the area ratio of thrombus to mouse cerebral venule. (C) The effect of CA on accumulative area ratio of thrombus to mouse cerebral venule during 60 min. ADP: the animals were treated with ADP at 0, 5, 15, and 25 min, respectively. CA + ADP: the animals were treated with CA (5 mg/kg) starting from 20 min before thrombus induction. Data are expressed as means ± SEM (n = 10). *p < 0.05 vs control group, #p < 0.05 vs ADP group.
Mentions: We next assessed the effect of CA on thrombus formation in mouse cerebral venules induced by ADP. As shown in Fig. 3A, only few of fluorescently labeled platelets (bright dots) were detected transiently adhered on the cerebral venular wall in all groups before the topical application of ADP (a1, b1, and c1), which persisted in the control group over the entire observation (a2–a7). In contrast, in ADP group, platelet adhesion was rapidly enhanced on the venular wall in mice, followed by a gradual build-up of platelet thrombus, in which fluorescently labeled platelets were seen as bright dots. The thrombus grew rapidly and obstructed partially the lumen of the vessel in a short time, developing to a comparable size to the venular diameter until it flowed downstream (b2–b4). The blood flow decreased as the thrombus grew, falling quickly over the last minutes and forming a stable thrombus (b5–b7). In the mice pretreated with CA (5 mg/kg), topical application of ADP-induced platelet adhesion and thrombus formation were markedly attenuated, as shown by the delayed thrombus formation and fewer accumulated platelets (c2–c4). After stopping ADP stimulation, thrombus disrupted into non-uniform fragments, which were quickly carried away by the blood flow (c5–c57).

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