Hyperlipidemia is a risk factor for clinically significant thrombotic events in cardiovascular diseases.Platelet reactivity in hyperlipidemic conditions is enhanced when platelet scavenger receptor CD36 recognizes oxi...Hyperlipidemia is a risk factor for clinically significant thrombotic events in cardiovascular diseases.Platelet reactivity in hyperlipidemic conditions is enhanced when platelet scavenger receptor CD36 recognizes oxidized lipids in oxidized low-density lipoprotein(ox-LDL)particles,a process that induces atherothrombosis.Sulforaphane(SFN)is a dietary isothiocyanate enriched in cruciferous vegetables and exerts multiple biological activities.The current study sought to investigate the efficacy of SFN on platelet hyperreactivity under hyperlipidemic conditions in vitro and in vivo.Using a series of platelet functional assays in human platelets in vitro,we demonstrated that SFN attenuated ox-LDL-increased platelet aggregation and activation(surface CD62P expression).Mechanistically,studies using pharmacological inhibitors clarified that these inhibitory effects of SFN were mainly modulated by down-regulating CD36-mediated activation of Src kinases,leading to enhanced activation of cyclic adenosine monophosphate/protein kinase A(cAMP/PKA)signaling,and resultant inhibition of NADPH oxidase 2(NOX2)-dependent generation of reactive oxygen species(ROS).Moreover,12-week supplementation of SFN-enriched broccoli sprout extract(BSE,0.06%diet)in hyperlipidemic C57BL/6J mice also decreased platelet hyperreactivity.Studies using pharmacological inhibitors of CD36,protein kinase A(PKA)and NOX2 showed that the efficacy of BSE supplementation was mainly through modulating CD36-mediated the cAMP/PKA/NOX2 signaling.Thus,through modulating the cAMP/PKA/NOX2 pathway and attenuating CD36-mediated platelet hyperreactivity,SFN may play important protective roles in atherothrombosis under hyperlipidemic conditions.展开更多
Platelet apoptosis is crucial for initiating atherothrombosis and is primarily induced by circulating oxidized lipids in oxidized low-density lipoprotein(ox-LDL)particles under hyperlipidemic conditions.Panax notogins...Platelet apoptosis is crucial for initiating atherothrombosis and is primarily induced by circulating oxidized lipids in oxidized low-density lipoprotein(ox-LDL)particles under hyperlipidemic conditions.Panax notoginseng has excellent medicinal and food dual-use characteristics.P.notoginseng saponins(PNS),the primary active ingredients extracted from the roots of P.notoginseng,are widely used as nutritional supplements owing to their multiple health benefits.In this study,we investigated the efficacy of PNS in platelet apoptosis under hyperlipidemic conditions in vitro and in vivo.In vitro assays demonstrated that PNS attenuated platelet mitochondrial dysfunction(loss of mitochondrial membrane potential)and subsequent apoptosis(caspase-9/-3 activation and phosphatidylserine exposure)in ox-LDL-stimulated human platelets.Mechanistically,PNS activated cAMP/PKA signaling,leading to decreased NOX2-mediated oxidative stress,and the resultant inhibition of platelet apoptosis.Moreover,12-week supplementation of PNS(0.02% diet)in hyperlipidemic C57BL/6J mice also significantly alleviated platelet mitochondrial dysfunction and apoptosis in vivo.Studies using pharmacological inhibitors of PKA and NOX2 demonstrated that the efficacy of PNS on platelet dysfunction in mice was primarily modulated by the up-regulation of cAMP/PKA signaling and the resultant inhibition of NOX2-dependent oxidative stress.Thus,these results suggest that PNS attenuate oxidative stress-mediated platelet apoptosis under hyperlipidemic conditions by up-regulating the cAMP/PKA signaling pathway in vitro and in vivo.Our study elucidated a possible novel strategy of involving the use of PNS to prevent atherothrombosis in hyperlipidemia.展开更多
基金supported by the National Natural Science Foundation of China(82003451 and 82003455)Yunnan Fundamental Research Projects(202101AT070033)the Start-Up Fund for Introduction of High-level Talents to Dali University(YBS2021015).
文摘Hyperlipidemia is a risk factor for clinically significant thrombotic events in cardiovascular diseases.Platelet reactivity in hyperlipidemic conditions is enhanced when platelet scavenger receptor CD36 recognizes oxidized lipids in oxidized low-density lipoprotein(ox-LDL)particles,a process that induces atherothrombosis.Sulforaphane(SFN)is a dietary isothiocyanate enriched in cruciferous vegetables and exerts multiple biological activities.The current study sought to investigate the efficacy of SFN on platelet hyperreactivity under hyperlipidemic conditions in vitro and in vivo.Using a series of platelet functional assays in human platelets in vitro,we demonstrated that SFN attenuated ox-LDL-increased platelet aggregation and activation(surface CD62P expression).Mechanistically,studies using pharmacological inhibitors clarified that these inhibitory effects of SFN were mainly modulated by down-regulating CD36-mediated activation of Src kinases,leading to enhanced activation of cyclic adenosine monophosphate/protein kinase A(cAMP/PKA)signaling,and resultant inhibition of NADPH oxidase 2(NOX2)-dependent generation of reactive oxygen species(ROS).Moreover,12-week supplementation of SFN-enriched broccoli sprout extract(BSE,0.06%diet)in hyperlipidemic C57BL/6J mice also decreased platelet hyperreactivity.Studies using pharmacological inhibitors of CD36,protein kinase A(PKA)and NOX2 showed that the efficacy of BSE supplementation was mainly through modulating CD36-mediated the cAMP/PKA/NOX2 signaling.Thus,through modulating the cAMP/PKA/NOX2 pathway and attenuating CD36-mediated platelet hyperreactivity,SFN may play important protective roles in atherothrombosis under hyperlipidemic conditions.
基金supported by Yunnan Fundamental Research Projects(202101AT070033202401AT070081)the National Natural Science Foundation of China(82260638).
文摘Platelet apoptosis is crucial for initiating atherothrombosis and is primarily induced by circulating oxidized lipids in oxidized low-density lipoprotein(ox-LDL)particles under hyperlipidemic conditions.Panax notoginseng has excellent medicinal and food dual-use characteristics.P.notoginseng saponins(PNS),the primary active ingredients extracted from the roots of P.notoginseng,are widely used as nutritional supplements owing to their multiple health benefits.In this study,we investigated the efficacy of PNS in platelet apoptosis under hyperlipidemic conditions in vitro and in vivo.In vitro assays demonstrated that PNS attenuated platelet mitochondrial dysfunction(loss of mitochondrial membrane potential)and subsequent apoptosis(caspase-9/-3 activation and phosphatidylserine exposure)in ox-LDL-stimulated human platelets.Mechanistically,PNS activated cAMP/PKA signaling,leading to decreased NOX2-mediated oxidative stress,and the resultant inhibition of platelet apoptosis.Moreover,12-week supplementation of PNS(0.02% diet)in hyperlipidemic C57BL/6J mice also significantly alleviated platelet mitochondrial dysfunction and apoptosis in vivo.Studies using pharmacological inhibitors of PKA and NOX2 demonstrated that the efficacy of PNS on platelet dysfunction in mice was primarily modulated by the up-regulation of cAMP/PKA signaling and the resultant inhibition of NOX2-dependent oxidative stress.Thus,these results suggest that PNS attenuate oxidative stress-mediated platelet apoptosis under hyperlipidemic conditions by up-regulating the cAMP/PKA signaling pathway in vitro and in vivo.Our study elucidated a possible novel strategy of involving the use of PNS to prevent atherothrombosis in hyperlipidemia.
