1. 中山大学公共卫生学院营养系,广东,广州,510080
2. 广东省营养与健康重点实验室,广东,广州,510080
3. 广东省营养转化工程技术研究中心,广东,广州,510080
4. 中山大学公共卫生学院(深圳),广东,深圳,518000
网络首发:2020-03-25,
纸质出版:2020
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凡蝶, 田泽众, 马熙麟, 等. 水溶性番茄浓缩物 Fruitflow 通过调控血小板 PI3K/Akt和MAPKs 信号通路抑制其活化、聚集及血栓形成[J]. 中山大学学报(医学科学版), 2020,41(2).
FAN Die, TIAN Ze-zhong, MA Xi-lin, et al. Fruitflow,a Water-soluble Tomato Concentrate,Inhibits Platelet Activation,Aggregation and Thrombosis by Regulating the Signaling Pathway of PI3K/Akt and MAPKs[J]. Journal of Sun Yat-sen University (Medical Sciences), 2020, 41(2).
凡蝶, 田泽众, 马熙麟, 等. 水溶性番茄浓缩物 Fruitflow 通过调控血小板 PI3K/Akt和MAPKs 信号通路抑制其活化、聚集及血栓形成[J]. 中山大学学报(医学科学版), 2020,41(2). DOI:
FAN Die, TIAN Ze-zhong, MA Xi-lin, et al. Fruitflow,a Water-soluble Tomato Concentrate,Inhibits Platelet Activation,Aggregation and Thrombosis by Regulating the Signaling Pathway of PI3K/Akt and MAPKs[J]. Journal of Sun Yat-sen University (Medical Sciences), 2020, 41(2). DOI:
【目的】血小板过度活化可促进血栓以及心血管疾病的发生发展,水溶性番茄浓缩物Fruitflow作为膳食补充剂在预防心血管疾病中发挥重要作用,但机制未明,且尚无Fruitflow对血栓形成影响的研究。本研究拟探讨Fruitflow对健康人血小板聚集、活化及血栓形成的影响,并探索其可能的作用机制。【方法】用不同浓度(0、20、40、80mg/L)的Fruitflow与健康成人血小板在体外共孵育,使用血小板聚集仪测定血小板聚集率;用流式细胞仪测定血小板表面αIIbβ3活化及纤维蛋白原结合率;用体视荧光显微镜观察FeCl3诱导的小鼠肠系膜动脉血栓模型中血栓的形成;通过小鼠剪尾实验测定出血时间;用Western blot检测血小板Akt、Erk1/2、JNK总蛋白表达及其磷酸化水平。【结果】在ADP诱导下,20、40、80mg/L的Fruitflow均可有效降低血小板聚集率,且效应呈剂量依赖性,其中80mg/L的Fruitflow干预组血小板聚集率从对照组的(43.75±5.91)%降到了(8.25±4.57)%(<0.001);与对照组(79.36±6.26)相比,Fruitflow明显抑制血小板αIIbβ3的活化,其中80mg/L剂量组降至(65.79±5.73,<0.01);与对照组(69.34±6.63)相比,仅80mg/L Fruitflow可明显降低血小板Fibrinogen binding百分比(56.70±8.68,<0.05);Fruitflow可抑制FeCl3诱导的肠系膜动脉血栓形成,80mg/L Fruitflow干预后血管堵塞时间由对照组的(22.50±4.86)延长至(39.20±4.61,<0.01),同时与对照组(5.95±0.69)相比,80mg/L Fruitflow干预组的出血时间(5.74±0.76)无明显变化(>0.05);Fruitflow下调血小板Akt、Erk1/2和JNK蛋白的磷酸化水平,与对照组相比,80mg/L Fruitflow干预后P-Akt/Akt由0.97±0.07降至0.33±0.13(<0.001),P-Erk1/2/Erk1/2由0.89±0.09降至0.24±0.02(<0.01),P-JNK/JNK由0.97±0.12降至0.45±0.04(<0.01)。【结论】抑制血小板PI3K/Akt和MAPKs信号可能是Fruitflow抑制血小板聚集活化和血栓形成的机制之一。
【Objective】Platelet hyper- reactivity plays an important role in thrombosis and cardiovascular diseases. As a dietary supplement,Fruitflow,a water-soluble tomato concentrate,plays an important role in preventing cardiovascular diseases,but the mechanisms have been poorly understood,and there is no direct evidence about the effect of Fruitflow on thrombosis. This study intends to explore the effects of Fruitflow on platelet aggregation,activation and thrombosis in healthy people,and explore its possible mechanism.【Methods】Platelets from healthy men and women were incubated with different concentrations(0,20,40,80 mg/L)of Fruitflow,and the platelet aggregation was measured by platelet aggregometer. Platelet αIIbβ3 activation and fibrinogen binding to the activated platelets was measured by flow cytometry. The thrombosis in FeCl3- induced mesenteric artery thrombosis model in mice was observed by stereoscopic fluorescence microscope. The bleeding time was measured by tail clipping in mice. The total and phosphorylation levels of platelet Akt, Erk1/2 and JNK were determined by Western blotting.【Results】After the stimulation of ADP,Fruitflow could significantly attenuate platelet aggregation in the dose of 20,40,80 mg/L. Compared with that in the control group(43.75±5.91)%, platelet aggregation in the 80 mg/L group decreased to(8.25 ± 4.57)%(< 0.000 1). Fruitflow also inhibited the activation of platelet αIIbβ3. Compared with that in the control group(79.36±6.26),the platelet αIIbβ3 in the 80 mg/L group decreased to(65.79±5.73, < 0.01). Fruitflow reduced the platelet fibrinogen binding to the activated platelets. Compared with that in the control group(69.34 ± 6.63),platelet fibrinogen binding to the activated platelets in the 80 mg/L group decreased to(56.70±8.86, < 0.05). Fruitflow attenuated the thrombosis in FeCl3-induced mesenteric artery thrombosis model. Compared with that in the control group(22.50±4.86),the vessel occlusion in the 80 mg/L group was extended to(39.20 ± 4.61,< 0.01). There was no statistical difference between the control group (5.95 ± 0.69) and the 80 mg/L group(5.74 ± 0.76)in bleeding time( > 0.05). Fruitflow down- regulated the phosphorylation levels of platelet Akt, Erk1/2 and JNK proteins. Compared with the control group,P- Akt/Akt decreased from 0.97 ± 0.07 to 0.33 ± 0.13( <0.001),P-Erk1/2/Erk1/2 decreased from 0.89±0.09 to 0.24±0.02( < 0.01),and P-JNK/JNK decreased from 0.97±0.12 to 0.45±0.04(< 0.01)in the 80 mg/L group.【Conclusion】Inhibition of platelet PI3K/Akt and MAPKs signaling pathway may be one of the mechanisms that Fruitflow inhibits platelet aggregation ,activation and thrombosis.
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