1.中山大学公共卫生学院(深圳),广东 深圳 518106
2.广东省营养膳食与健康重点实验室,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,广东 广州 510080
李孔耀,硕士生,研究方向:营养与慢性病的防治,E-mail:liky33@mail2.sysu.edu.cn
收稿:2021-01-22,
纸质出版:2021-05-20
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李孔耀,施译琳,马熙麟等.水溶性番茄浓缩物经自噬途径调控血小板的氧化损伤[J].中山大学学报(医学科学版),2021,42(03):321-327.
LI Kong-yao,SHI Yi-lin,MA Xi-lin,et al.Fruitflow, a Water-soluble Tomato Extract, Regulates Platelet Oxidative Damage via Autophagy in Vitro[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(03):321-327.
李孔耀,施译琳,马熙麟等.水溶性番茄浓缩物经自噬途径调控血小板的氧化损伤[J].中山大学学报(医学科学版),2021,42(03):321-327. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0101.
LI Kong-yao,SHI Yi-lin,MA Xi-lin,et al.Fruitflow, a Water-soluble Tomato Extract, Regulates Platelet Oxidative Damage via Autophagy in Vitro[J].Journal of Sun Yat-sen University(Medical Sciences),2021,42(03):321-327. DOI: 10.13471/j.cnki.j.sun.yat-sen.univ(med.sci).2021.0101.
目的
2
过度氧化应激可促进血小板氧化损伤,从而加快心血管疾病的发生发展。水溶性番茄浓缩物(Fruitflow)具有抑制血小板聚集、活化的作用,但是否通过自噬减轻血小板氧化损伤仍未见报道。本研究拟通过体外实验探讨Fruitflow对过氧化氢(H
2
O
2
)导致的血小板氧化损伤的影响及其潜在机制。
方法
2
用不同剂量的Fruitflow (0、20、40、80 mg/L)与纯化血小板预孵育30 min,随后加入H
2
O
2
共孵育60 min。用流式细胞术测定线粒体膜电位(ΔΨm)去极化的水平;用酶标仪测定活性氧(ROS)生成水平;用Western blot测定p53、phospho-p53、LC3 Ⅱ/Ⅰ、p62的表达水平。
结果
2
Fruitflow 显著减轻H
2
O
2
导致的血小板ΔΨm 去极化(
P
<
0.05),ROS 生成和p53磷酸化(
P
<
0.05)。自噬抑制剂3-甲基腺嘌呤(3-Methyladenine,3-MA)可以显著逆转Fruitflow 对H
2
O
2
处理血小板ROS生成的抑制作用(
P
<
0.05)。Fruitflow 可以降低静息血小板p62的表达、升高LC3Ⅱ/Ⅰ的比例(
P
<
0.05),也显著升高H
2
O
2
处理的血小板LC3Ⅱ/Ⅰ的表达水平(
P
<
0.05)。此外,3-MA显著逆转Fruitflow降低H
2
O
2
处理的血小板ΔΨm 去极化的作用(
P
<
0.05)。
结论
2
Fruitflow可在体外通过促进血小板自噬,显著减轻H
2
O
2
导致的血小板氧化损伤。
Objective
2
Excessive oxidative stress can promote platelet oxidative damage and plays an important role in the occurrence and development of cardiovascular diseases. Fruitflow, a water-soluble tomato extract, can inhibit platelet aggregation and activation, but it has not been reported whether Fruitflow can reduce platelet oxidative damage through autophagy. Therefore, we conducted this study to investigate the effect of Fruitflow on oxidative damage in H
2
O
2
-treated platelets as well as the underlying mechanism
in vitro
.
Methods
2
Gel-filtered platelets from healthy people were pre-incubated with different concentrations (0, 20, 40, 80 mg/L) of Fruitflow for 30 minutes, and then treated with H
2
O
2
(1 mmol/L) for another 60 minutes. The level of mitochondrial membrane potential(ΔΨm)depolarization was determined by flow cytometry. Enzyme-linked immunosorbent assay was used to detect reactive oxygen species (ROS) level. The expression of p53, phospho-p53, LC3Ⅱ/Ⅰ, and p62 was detected by Western blot.
Results
2
Fruitflow significantly inhibited the ΔΨm depolarization, ROS production and p53 phosphorylation in H
2
O
2
-treated platelets (
P
<
0.05). The autophagy inhibiter (3-MA) significantly reversed the effect of Fruitflow on reduction of platelet ROS generation (
P
<
0.05). In addition, Fruitflow reduced p62 and increased the expression level of LC3Ⅱ/Ⅰ in resting platelets (
P
<
0.05), and increased the expression level of LC3Ⅱ/Ⅰ in H
2
O
2
-treated platelets (
P
<
0.05). Furthermore, the effect of Fruitflow on decreasing ΔΨm depolarization in H
2
O
2
-treated platelet was reversed by 3-MA (
P
<
0.05).
Conclusion
2
Fruitflow can significantly reduce the H
2
O
2
-induced platelet oxidative damage by promoting autophagy
in vitro
.
Van Der Meijden PEJ , Heemskerk JWM . Platelet biology and functions: new concepts and clinical perspectives [J]. Nat Rev Cardiol , 2019 , 16 ( 3 ): 166 - 179 .
Wang L , Wu Q , Fan Z , et al . Platelet mitochondrial dysfunction and the correlation with human diseases [J]. Biochem Soc Trans , 2017 , 45 ( 6 ): 1213 - 1223 .
Tang WH , Stitham J , Jin Y , et al . Aldose reductase-mediated phosphorylation of p53 leads to mitochondrial dysfunction and damage in diabetic platelets [J]. Circulation , 2014 , 129 ( 15 ): 1598 - 1609 .
Luo XL , Jiang JY , Huang Z , et al . Autophagic regulation of platelet biology [J]. J Cell Physiol , 2019 , 234 ( 9 ): 14483 - 14488 .
Mizushima N , Komatsu M . Autophagy: renovation of cells and tissues [J]. Cell , 2011 , 147 ( 4 ): 728 - 741 .
Yun HR , Jo YH , Kim J , et al . Roles of autophagy in oxidative stress [J]. Int J Mol Sci , 2020 , 21 ( 9 ): 3289 .
Feng W , Chang C , Luo D , et al . Dissection of autophagy in human platelets [J]. Autophagy , 2014 , 10 ( 4 ): 642 - 651 .
Lee SH , Du J , Stitham J , et al . Inducing mitophagy in diabetic platelets protects against severe oxidative stress [J]. EMBO Mol Med , 2016 , 8 ( 7 ): 779 - 795 .
Brandhorst S , Longo VD . Dietary restrictions and nutrition in the prevention and treatment of cardiovascular disease [J]. Circ Res , 2019 , 124 ( 6 ): 952 - 965 .
Cheng HM , Koutsidis G , Lodge JK , et al . Tomato and lycopene supplementation and cardiovascular risk factors: A systematic review and meta-analysis [J]. Atherosclerosis , 2017 , 257 : 100 - 108 .
O'Kennedy N , Raederstorff D , Duttaroy AK . Fruitflow: the first European Food Safety Authority-approved natural cardio-protective functional ingredient [J]. Eur J Nutr , 2017 , 56 ( 2 ): 461 - 482 .
Uddin M , Biswas D , Ghosh A , et al . Consumption of Fruitflow((R)) lowers blood pressure in pre-hypertensive males: a randomised, placebo controlled, double blind, cross-over study [J]. Int J Food Sci Nutr , 2018 , 69 ( 4 ): 494 - 502 .
Guo Q , Li F , Duan Y , et al . Oxidative stress, nutritional antioxidants and beyond [J]. Sci China Life Sci , 2020 , 63 ( 6 ): 866 - 874 .
汪玉芳 , 贺瑞坤 , 张旭光 . 水溶性番茄浓缩物抗氧化和辅助降血脂功效研究 [J]. 食品安全质量检测学报 , 2020 , 11 ( 13 ): 4531 - 4535 .
Wang YF , He RK , Zhang XG . Study on the antioxidation and assistant hypolipidemic function of water soluble tomato concentrate [J]. J Food Safety & Quality , 2020 , 11 ( 13 ): 4531 - 4535 .
Yang Y , Shi Z , Reheman A , et al . Plant food delphinidin-3-glucoside significantly inhibits platelet activation and thrombosis: novel protective roles against cardiovascular diseases [J]. PloS one , 2012 , 7 ( 5 ): e37323 .
Khosravi M , Poursaleh A , Ghasempour G , et al . The effects of oxidative stress on the development of atherosclerosis [J]. Bio Chem , 2019 , 400 ( 6 ): 711 - 732 .
El Hauari M . Platelet oxidative stress and its relationship with cardiovascular diseases in type 2 diabetes mellitus patients [J]. Curr Med Chem , 2019 , 26 ( 22 ): 4145 - 4165 .
Khodadi E . Platelet function in cardiovascular disease: Activation of molecules and activation by molecules [J]. Cardiovasc Toxicol , 2020 , 20 ( 1 ): 1 - 10 .
Hada Y , Uchida HA , Otaka N , et al . The protective effect of chlorogenic acid on vascular senescence via the Nrf2/HO-1 pathway [J]. Int J Mol Sci , 2020 , 21 ( 12 ): 4527 .
Ma JQ , Liu CM , Yang W . Protective effect of rutin against carbon tetrachloride-induced oxidative stress, inflammation and apoptosis in mouse kidney associated with the ceramide, MAPKs, p53 and calpain activities [J]. Chem Biol interact , 2018 , 286 : 26 - 33 .
Miyamoto S . Autophagy and cardiac aging [J]. Cell Death Differ , 2019 , 26 ( 4 ): 653 - 664 .
Ma Y , Yang L , Ma J , et al . Rutin attenuates doxorubicin-induced cardiotoxicity via regulating autophagy and apoptosis [J]. Biochim Biophys Acta Mol Basis Dis , 2017 , 1863 ( 8 ): 1904 - 1911 .
Guo W , Liu J , Zhang Y , et al . Dehydroandrographolide inhibits mastitis by activating autophagy without affecting intestinal flora [J]. Aging (Albany NY) , 2020 , 12 ( 14 ): 14050 - 14065 .
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 ): 243 - 250 .
Fan D , Tian ZZ , Ma XL , 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]. J Sun Yat-sen Univ (Med Sci) , 2020 , 41 ( 2 ): 243 - 250 .
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