多柔比星、二甲双胍对干细胞心肌样分化的代谢组分研究
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摘要
目的结合代谢组学技术和胚胎干细胞心肌定向诱导分化方法,对多柔比星和二甲双胍造成代谢组分的变化进行研究。方法以小鼠胚胎干细胞体外心肌定向诱导分化为模型,分化培养液中添加不同剂量的多柔比星(3.40、0.17μmol·L~(-1))以及二甲双胍(905.7μmol·L~(-1))。通过对诱导形成拟胚体的一般观察,统计自主节律跳动、拟胚体的数量、频率等;免疫细胞化学法检测心肌特异性蛋白—心肌肌钙蛋白T(cTnT)的表达;应用代谢组学技术初步筛选差异代谢图谱,以多柔比星和二甲双胍共同影响的代谢组分为目标,尝试寻找心脏毒性敏感标志物。结果一般观察和免疫细胞化学结果显示,多柔比星高浓度明显抑制向心肌细胞分化,cTnT表达明显增加,低浓度组拟胚体自主节律明显增快,cTnT表达变化不明显;二甲双胍未见明显变化。代谢组学结果显示,多柔比星和二甲双胍可致细胞内同型半胱氨酸(Hcy)、铁卟啉(FeTMPyP)、叶酸、半胱氨酸(Cys)、cAMP等组分明显降低。结论氧化应激在心脏分化发育过程中起重要作用,Hcy、FeTMPyP、Cys等具有成为心脏毒性敏感标志物的潜力。
Aim To analyze the changes of the metabolic profiling of cardiomyocyte-like cells via introducing metabonomics and stem cell differentiation tech-nology. Methods The mouse embryonic stem cells
Aim To analyze the changes of the metabolic profiling of cardiomyocyte-like cells via introducing metabonomics and stem cell differentiation tech-nology. Methods The mouse embryonic stem cells
引文
[1] 李 黎,申秀萍,钱 鑫,等. 体外心肌毒性代谢组学研究中模型药物多柔比星浓度及受试时间筛选[J]. 药物评价研究,2016,39(3):398-402.
[1] Li L,Shen X P,Qian X,et al. Selection of test concentrations and test times of model drugs for metabonomic studies of in vitro cardiotoxicity[J]. Drug Eval Res, 2016,39(3):398-402.
[2] 顾觉奋. 多柔比星毒副作用机制及其防治研究进展[J]. 国外医药抗生素分册, 2017,38(4):145-51.
[2] Gu J F. Current views on the mechanism of doxorubicin-induced cardiotoxicity and its prevention[J]. World Notes Antibiot, 2017,38(4):145-51.
[3] 徐鸿洁,魏大军. 蒽环类化疗药物心脏毒性及其治疗研究进展[J]. 中国城乡企业卫生, 2015(4):37-9.
[3] Xu H J,Wei D J. Progress on anthracycline cardiotoxicity and treatment[J]. Chin J Urban Rural Ind Hyg, 2015(4):37-9.
[4] 吴运香,张 野,谢春林,等. SD大鼠阿霉素慢性心力衰竭模型的建立与评价[J]. 中国药理学通报, 2011,27(8): 1170-3.
[4] Wu Y X,Zhang Y,Xie C L,et al. Establishment and evaluation of adriamycin-induced chronic heart failure model in SD rats[J]. Chin Pharmacol Bull, 2011,27(8):1170-3.
[5] 孙凤姣,张译丹,张密霞,等. β-AR/PKA/CaMK Ⅱ信号通路在阿霉素诱导大鼠心肌细胞凋亡中的作用[J]. 中国药理学通报, 2017,33(3):360-5.
[5] Sun F J,Zhang Y D,Zhang M X,et al. Roles of β-AR/PKA/CaMK Ⅱ signaling pathway in cardiomyocyte apoptosis induced by adriamycin in rats[J]. Chin Pharmacol Bull, 2017,33(3):360-5.
[6] Anderson A B,Arriaga E A. Subcellular metabolite profiles of the parent CCRF-CEM and the derived CEM/C2 cell lines after treatment with doxorubicin[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2004,808(2):295-302.
[7] 从文娟,刘清飞,梁琼麟,等. 基于血清代谢组学的吡柔比星注射剂的累积毒性作用研究[J]. 中国药理学通报, 2012,28(9):1294-9.
[7] Cong W J,Liu Q F,Liang Q L,et al. Serum metabolites of commerical pirarubicin injections[J]. Chin Pharmacol Bull, 2012,28(9): 1294-9.
[8] Kang J L,Lee H S,Pack I S,et al. Iron tetrakis(n-methyl-4′-pyridyl) porphyrinato (FeTMPyP) is a potent scavenging antioxidant and an inhibitor of stimulant-induced NF-kappaB activation of RAW 264.7 macrophages[J]. J Toxicol Environ Health A,2001,64(4):291-310.
[9] Nawara K, Beeckman H,Krysiński P,Blanchard G J. Structure-dependent complexation of Fe3+ by anthracyclines. 2. The roles of methoxy and daunosamine functionalities[J]. J Phys Chem B,2013,117(23):6868-73.
[10] 彭 琳,唐小海,严 伟,等. 果胶阿霉素对大鼠心脏毒性的研究[J]. 中国药理学通报, 2016,32(8):1075-80.
[10] Peng L,Tang X H,Yan W,et al. Study of pectin-adriamycin conjugate to cardiac toxicity in rats[J]. Chin Pharmacol Bull, 2016,32(8):1075-80.
[11] 阳冠明,孙胜涛,李树全,等. β-胡萝卜素对阿霉素致大鼠心肌组织的超氧化物歧化酶、谷胱甘肽过氧化物酶mRNA表达改变的影响[J]. 中国药理学通报,2006,22(4): 465-70.
[11] Yang G M,Sun S T,Li S Q,et al. Effect of beta-carotene on adriamycin induced changes of expression of mRNA of superoxide dismutase and glutathione peroxidase in myocardial tissue of rats[J]. Chin Pharmacol Bull,2006,22(4):465-70.
[12] 张 辰,骆海燕,高小博,等. 叶酸对阿霉素诱导心脏毒性的拮抗作用研究[J]. 中国计划生育学杂志,2017,25(3):159-63.
[12] Zhang C,Luo H Y,Gao X B,et al. The study of folic acid against doxorubicin-induced cardiotoxicity[J]. Chin J Fam Plann, 2017,25(3):159-63.
[13] Sahu B D,Kumar J M,Kuncha M,et al. Baicalein alleviates doxorubicin-induced cardiotoxicity via suppression of myocardial oxidative stress and apoptosis in mice[J]. Life Sci, 2016,144:8-18.
[14] Zhou S,Starkov A,Froberg M K,et al. Cumulative and irreversible cardiac mitochondrial dysfunction induced by doxorubicin[J]. Cancer Res,2001,61(2):771-7.
[15] 薛朝军,刘克辛. 二甲双胍抗肿瘤机制的研究进展[J]. 药学学报, 2015,50(10):1210-6.
[15] Xue C J,Liu K X. Advances of the anti-tumor research of metformin[J]. Acta Pharm Sin, 2015,50(10):1210-6.
[1] Li L,Shen X P,Qian X,et al. Selection of test concentrations and test times of model drugs for metabonomic studies of in vitro cardiotoxicity[J]. Drug Eval Res, 2016,39(3):398-402.
[2] 顾觉奋. 多柔比星毒副作用机制及其防治研究进展[J]. 国外医药抗生素分册, 2017,38(4):145-51.
[2] Gu J F. Current views on the mechanism of doxorubicin-induced cardiotoxicity and its prevention[J]. World Notes Antibiot, 2017,38(4):145-51.
[3] 徐鸿洁,魏大军. 蒽环类化疗药物心脏毒性及其治疗研究进展[J]. 中国城乡企业卫生, 2015(4):37-9.
[3] Xu H J,Wei D J. Progress on anthracycline cardiotoxicity and treatment[J]. Chin J Urban Rural Ind Hyg, 2015(4):37-9.
[4] 吴运香,张 野,谢春林,等. SD大鼠阿霉素慢性心力衰竭模型的建立与评价[J]. 中国药理学通报, 2011,27(8): 1170-3.
[4] Wu Y X,Zhang Y,Xie C L,et al. Establishment and evaluation of adriamycin-induced chronic heart failure model in SD rats[J]. Chin Pharmacol Bull, 2011,27(8):1170-3.
[5] 孙凤姣,张译丹,张密霞,等. β-AR/PKA/CaMK Ⅱ信号通路在阿霉素诱导大鼠心肌细胞凋亡中的作用[J]. 中国药理学通报, 2017,33(3):360-5.
[5] Sun F J,Zhang Y D,Zhang M X,et al. Roles of β-AR/PKA/CaMK Ⅱ signaling pathway in cardiomyocyte apoptosis induced by adriamycin in rats[J]. Chin Pharmacol Bull, 2017,33(3):360-5.
[6] Anderson A B,Arriaga E A. Subcellular metabolite profiles of the parent CCRF-CEM and the derived CEM/C2 cell lines after treatment with doxorubicin[J]. J Chromatogr B Analyt Technol Biomed Life Sci,2004,808(2):295-302.
[7] 从文娟,刘清飞,梁琼麟,等. 基于血清代谢组学的吡柔比星注射剂的累积毒性作用研究[J]. 中国药理学通报, 2012,28(9):1294-9.
[7] Cong W J,Liu Q F,Liang Q L,et al. Serum metabolites of commerical pirarubicin injections[J]. Chin Pharmacol Bull, 2012,28(9): 1294-9.
[8] Kang J L,Lee H S,Pack I S,et al. Iron tetrakis(n-methyl-4′-pyridyl) porphyrinato (FeTMPyP) is a potent scavenging antioxidant and an inhibitor of stimulant-induced NF-kappaB activation of RAW 264.7 macrophages[J]. J Toxicol Environ Health A,2001,64(4):291-310.
[9] Nawara K, Beeckman H,Krysiński P,Blanchard G J. Structure-dependent complexation of Fe3+ by anthracyclines. 2. The roles of methoxy and daunosamine functionalities[J]. J Phys Chem B,2013,117(23):6868-73.
[10] 彭 琳,唐小海,严 伟,等. 果胶阿霉素对大鼠心脏毒性的研究[J]. 中国药理学通报, 2016,32(8):1075-80.
[10] Peng L,Tang X H,Yan W,et al. Study of pectin-adriamycin conjugate to cardiac toxicity in rats[J]. Chin Pharmacol Bull, 2016,32(8):1075-80.
[11] 阳冠明,孙胜涛,李树全,等. β-胡萝卜素对阿霉素致大鼠心肌组织的超氧化物歧化酶、谷胱甘肽过氧化物酶mRNA表达改变的影响[J]. 中国药理学通报,2006,22(4): 465-70.
[11] Yang G M,Sun S T,Li S Q,et al. Effect of beta-carotene on adriamycin induced changes of expression of mRNA of superoxide dismutase and glutathione peroxidase in myocardial tissue of rats[J]. Chin Pharmacol Bull,2006,22(4):465-70.
[12] 张 辰,骆海燕,高小博,等. 叶酸对阿霉素诱导心脏毒性的拮抗作用研究[J]. 中国计划生育学杂志,2017,25(3):159-63.
[12] Zhang C,Luo H Y,Gao X B,et al. The study of folic acid against doxorubicin-induced cardiotoxicity[J]. Chin J Fam Plann, 2017,25(3):159-63.
[13] Sahu B D,Kumar J M,Kuncha M,et al. Baicalein alleviates doxorubicin-induced cardiotoxicity via suppression of myocardial oxidative stress and apoptosis in mice[J]. Life Sci, 2016,144:8-18.
[14] Zhou S,Starkov A,Froberg M K,et al. Cumulative and irreversible cardiac mitochondrial dysfunction induced by doxorubicin[J]. Cancer Res,2001,61(2):771-7.
[15] 薛朝军,刘克辛. 二甲双胍抗肿瘤机制的研究进展[J]. 药学学报, 2015,50(10):1210-6.
[15] Xue C J,Liu K X. Advances of the anti-tumor research of metformin[J]. Acta Pharm Sin, 2015,50(10):1210-6.