Proteomic changes by radio-mitigative thrombopoietin receptor agonist romiplostim in the blood of mice exposed to lethal total-body irradiation
Proteomic changes by radio-mitigative thrombopoietin receptor agonist romiplostim in the blood of mice exposed to lethal total-body irradiation
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DOI：
10.1080/09553002.2020.1787546
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发表时间：
2020-07-10
影响因子：
2.6
通讯作者：
Kashiwakura, Ikuo
中科院分区：
文献类型：
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作者：
Nishida, Teruki;Yamaguchi, Masaru;Kashiwakura, Ikuo
目的血小板生成素受体激动剂罗米普罗辛(romiplostin，RP)是一种治疗免疫性血小板减少症的药物，可使接受致死剂量电离辐射的小鼠完全存活。放射防护/缓解效应的估计机制已被提出；然而，具体的作用机制仍不清楚。本研究通过蛋白质组学的方法分析了血液中蛋白质的变化，旨在阐明RP的辐射防护/缓解作用的机制。材料与方法8周龄雌性C57BL/6J小鼠随机分为5组：0d对照组，10d和18d全身照射组，10d和18d全身照射+RP组，每组3只，接受7Gy137Csγ射线照射，剂量率为0.74Gymin。从伤后2小时开始，给小鼠腹腔注射RP，剂量为50微克/公斤，每日1次，连续3天。分别于伤后第10天和第18天采集小鼠血清，用液相色谱-串联质谱仪进行分析。结果用蛋白质组学方法从致死剂量脑损伤小鼠体内检测到9种蛋白质：角蛋白、II型细胞骨架1、果糖-1，6-二磷酸酶、胞浆10-甲酰基四氢叶酸脱氢酶、肽基-脯氨酸顺式-反式异构酶A、甘氨酸N-甲基转移酶、谷胱甘肽-S-转移酶MU-1、累古霉素、果糖-二磷酸酯酶B和甜菜碱-同型半胱氨酸-S甲基转移酶。在脑损伤后第10天，脑损伤组KRT1的表达显著高于对照组(P<0.05)，而脑损伤+RP组的KRT1表达明显受到抑制(P<0.05)。同样，脑损伤后第18天检测到的其他8种蛋白的表达水平在脑损伤组显著高于脑损伤组(4.29~27.44倍)，而脑损伤加RP组显著低于脑损伤组(P<0.05)。结论用蛋白质组学方法从致死剂量脑损伤小鼠体内检测到的269种蛋白质中鉴定出9种蛋白质。这些蛋白质也有望成为高剂量辐射损伤的指标。
Purpose The thrombopoietin receptor agonist romiplostim (RP) is a therapeutic agent for immune thrombocytopenia that can achieve complete survival in mice exposed to a lethal dose of ionizing radiation. The estimated mechanism of the radio-protective/mitigative effects of RP has been proposed; however, the detailed mechanism of action remains unclear. This study aimed to elucidate the mechanism of the radio-protective/mitigative effects of RP, the fluctuation of protein in the blood was analyzed by proteomics. Materials and Methods Eight-week-old female C57BL/6J mice were randomly divided into 5 groups; control at day 0, total-body irradiation (TBI) groups at day 10 and day 18, and TBI plus RP groups at day 10 and day18, consisting of 3 mice per group, and subjected to TBI with 7 Gy of(137)Cs gamma-rays at a dose rate of 0.74 Gy/min. RP was administered intraperitoneally to mice at a dose of 50 mu g/kg once daily for 3 days starting 2 hours after TBI. On day 10 and day 18 after TBI, serum collected from each mouse was analyzed by liquid chromatography tandem mass spectrometry. Results Nine proteins were identified by proteomics methods from 269 analyzed proteins detected in mice exposed to a lethal dose of TBI: keratin, type II cytoskeletal 1 (KRT1), fructose-1, 6-bisphosphatase (FBP1), cytosolic 10-formyltetrahydrofolate dehydrogenase (ALDH1L1), peptidyl-prolyl cis-trans isomerase A (PPIA), glycine N-methyltransferase (GNMT), glutathione S-transferase Mu 1 (GSTM1), regucalcin (RGN), fructose-bisphosphate aldolase B (ALDOB) and betain-homocysteine S-methyltransferase 1 (BHMT). On the 10th day after TBI, KRT1 was significantly increased (p < 0.05) by 4.26-fold compared to the control group in the TBI group and significantly inhibited in the TBI plus RP group (p < 0.05). Similarly, the expression levels of other 8 proteins detected at 18th day after TBI were significantly increased by 4.29 to 27.44-fold in the TBI group, but significantly decreased in the TBI plus RP group compared to the TBI group, respectively. Conclusion Nine proteins were identified by proteomics methods from 269 analyzed proteins detected in mice exposed to a lethal dose of TBI. These proteins are also expected to be indicators of the damage induced by high-dose radiation.