硫前列酮影响小鼠代谢的NMR研究

波谱学杂志

硫前列酮影响小鼠代谢的NMR研究

严星^1,2，张利民¹，安艳捧^1,2，李洪德^1,2，唐惠儒^1*

1. 波谱与原子分子物理国家重点实验室（中国科学院武汉物理与数学研究所），湖北武汉 430071； 
2.中国科学院大学，北京 100049

收稿日期:2012-04-27 修回日期:2012-05-08 出版日期:2013-03-05 发布日期:2013-03-05
通讯作者: 唐惠儒，电话：027-87198430，huiru.tang@wipm.ac.cn E-mail:huiru.tang@wipm.ac.cn
作者简介:严星（1986-），男，湖北武汉人，硕士研究生，主要从事基于核磁共振的代谢组学研究. 电话：027-87198430，E-mail：yanxing2008049@163.com
基金资助:
国家重点基础研究发展计划（“973”计划）资助项目（2010CB912501）.

NMR-Based Metabonomic Analysis for Sulprostone-Induced Mice

YAN Xing^1,2，ZHANG Li-min¹，AN Yan-peng^1,2，LI Hong-de^1,2，TANG Hui-ru^1*

1. State Key Laboratory of Magnetic Resonance and Atomic and Molecular Physics, (Wuhan Institute of Physics and Mathematics, Chinese Academy of Sciences), Wuhan 430071, China;
2. University of Chinese Academy of Sciences, Beijing 100049, China

Received:2012-04-27 Revised:2012-05-08 Online:2013-03-05 Published:2013-03-05
Contact: TANG Hui-ru，Tel: 027-87198430，huiru.tang@wipm.ac.cn E-mail:huiru.tang@wipm.ac.cn
About author:严星（1986-），男，湖北武汉人，硕士研究生，主要从事基于核磁共振的代谢组学研究. 电话：027-87198430，E-mail：yanxing2008049@163.com
Supported by:
国家重点基础研究发展计划（“973”计划）资助项目（2010CB912501）.

摘要/Abstract

摘要：

前列腺素E2（PGE2）作为一种体内广泛分布的活性物质，通过与其特异性受体EP1，EP2，EP3和EP4结合实现信号跨膜转导，参与许多重要的生理病理过程. 硫前列酮作为PGE2的类似物，通过激活EP1和EP3受体发挥其生理作用，但相关的代谢基础还不甚清楚. 该研究运用基于核磁共振（NMR）技术的代谢组学方法研究了EP1和EP3受体激活剂硫前列酮对小鼠血清和肝脏代谢组的影响. 结果表明，中高剂量硫前列酮处理32天会导致小鼠肝脏代谢组中的烟酰胺腺嘌呤二核苷酸磷酸、烟酰胺腺嘌呤二核苷酸、二磷酸尿苷、磷酸腺苷和胆汁酸的明显增加，同时肝糖原、葡萄糖、苯丙氨酸、酪氨酸、尿苷、肌苷、烟碱酸和氧化型谷胱甘肽明显减少. 恢复3周后，高剂量硫前列酮处理小鼠会导致肝脏代谢组中的胆碱水平比对照组高. 这些结果表明EP1和EP3受体激活剂硫前列酮会对小鼠肝脏的糖、核酸和氨基酸等代谢产生影响. 同时，未发现硫前列酮对血清代谢组产生明显影响，这可能与血液循环系统维持机体内环境相对稳定有关. 以上研究结果为认识PGE2-EP1/3信号通路在代谢中的作用提供了基础数据.

关键词: 核磁共振(NMR), 硫前列酮, 代谢组, 前列腺素E2

Abstract:

Prostaglandin E2 (PGE2) is a widely distributed substance in body, and involved in many physiological and pathological processes. It regulates signal transduction pathways through binding to EP1, EP2, EP3 and EP4 receptors. PGE2 is also. Sulprostone is an analogue of PGE2, playing its physiological functions through activation of EP1 and EP3. The metabolic foundation of PGE2 is still poorly understood. This work studied the metabonome of serum and liver in sulprostone-induced mice using nuclear magnetic resonance (NMR)-based analysis. The results showed that following 32 days of sulprostone treatment (i.e., 0.1 mg/kg, medium dose; 1 mg/kg, high dose), the animals showed significantly increased NADP⁺，NAD⁺，UDP，AMP and bile acids levels, and significantly reduced levels of glycogen, glucose, phenylalanine, tyrosine, uridine, inosine, nicotinic acid and GSSG in the liver. After three-weeks of recovery, choline elevation persisted in the high dose group. These results indicated that sulprostone caused metabolic disturbance of glucose, nucleic acid and amino acids in the liver. No sulprostoneinduced changes were observed in the serum data, probably due to hemeostasis. The results obtained provided more insights into the PGE2-EP1/3-signaling pathways in mouse, especially in the metabolic aspects.

Key words: NMR, sulprostone, metabonome, prostaglandin E2

中图分类号:

O482.53

严星，张利民，安艳捧，李洪德，唐惠儒*. 硫前列酮影响小鼠代谢的NMR研究[J]. 波谱学杂志.

YAN Xing，ZHANG Li-min，AN Yan-peng，LI Hong-de, TANG Hui-ru*. NMR-Based Metabonomic Analysis for Sulprostone-Induced Mice[J]. Chinese Journal of Magnetic Resonance.

参考文献

[1] Thorburn G D. The placenta, prostaglandins and parturition: a review[J]. Reprod Fert Dev, 1991, 3(3): 277-294.

[2] Sugimoto Y, Narumiya S, Ichikawa A. Distribution and function of prostanoid receptors: studies from knockout mice[J]. Prog Lipid Res, 2000, 39(4): 289-314.

[3] Narumiya S, Sugimoto Y, Ushikubi F. Prostanoid receptors: structures, properties, and functions[J]. Phys Rev, 1999, 79(4): 1 193-1 226.

[4] Kreydiyyeh S I, Riman S, Serhan M, et al. TNF-alpha modulates hepatic Na⁺K⁺ATPase activity via PGE2 and EP2 receptors[J]. Prostag Oth Lipid Med, 2007, 83(4): 295-303.

[5] Kimura M, Osumi S, Ogihara M. Prostaglandin E-2 (EP1) receptor agonist-induced DNA synthesis and proliferation in primary cultures of adult rat hepatocytes: The involvement of TGF-alpha[J]. Endocrinology, 2001, 142(10): 4 428-4 440.

[6] Mater M K, Thelen A P, Jump D B. Arachidonic acid and PGE(2) regulation of hepatic lipogenic gene expression[J]. J Lipid Research, 1999, 40(6): 1 045-1 052.

[7] Kuhnz W, Hoyer G A, Backhus S. Identification of the major metabolites of the prostaglandin E2-analogue sulprostone in human plasma, and isolation from urine (in vivo) and liver perfusate (in vitro) of female guinea-pigs[J]. Drug Metab Dispos, 1991, 19(5): 920-925.

[8] Liu Cheng-quan(刘承权), Chen Gen-fu(陈根富), Hu Yan(胡炎), et al. A pharmacodynamic study on sulprostone(磺前列酮的药效学研究)[J]. Chin Pharmacol Bull(中国药理学通报), 1992, 6(1): 471-474.

[9] Zhao Xiu-ju（赵秀举），Wang Yu-lan（王玉兰）. Applications of NMR-based metabonomic approaches in the assessment of drug toxicity（代谢组NMR分析与药物毒理研究）[J]. Chinese J Magn Reson（波谱学杂志） 2011, 28(1): 1-17.

[10] Waters N J, Garrod S, Farrant R D, et al. High-resolution magic angle spinning ¹]H NMR spectroscopy of intact liver and kidney: Optimization of sample preparation procedures and biochemical stability of tissue during spectral acquisition[J]. Anal Biochem, 2000, 282(1): 16-23.

[11] vanBaal J, deJong M D, Zijlstra F J, et al. Endogenously produced prostanoids stimulate calcium reabsorption in the rabbit cortical collecting system[J]. J Physiol-London, 1996, 497(1): 229-239.

[12] Liu M L, Nicholson J K, London J C. High-resolution diffusion and relaxation edited one- and two-dimensional ¹H NMR- spectroscopy of biological fluids[J]. Anal Chem, 1996, 68(19): 3 370-3 376.

[13] Tang H R, Wang Y L, Nicholson J K. Use of relaxation-edited one-dimensional and two dimensional nuclear magnetic resonance spectroscopy to improve detection of small metabolites in blood plasma[J]. Anal Biochem, 2004, 325(2): 260-272.

[14] Xiao Chao-ni（肖超妮），Liu Hong-bing（刘红兵），Dai Hui（戴惠），et al. Efficient Identification and structural elucidation of metabolites using HPLC-DAD-SPE-CryoNMRMS method（基于HPLC-DAD-SPE-CryoNMR-MS技术的代谢物快速定性和结构确定）[J]. Chinese J Magn Reson（波谱学杂志）, 2009, 26(1): 1-16.

[15] Zhang L M, Ye Y F, An Y P, et al. Systems responses of rats to aflatoxin B1 exposure revealed with metabonomic changes in multiple biological matrices[J]. J Proteome Res, 2011, 10(2): 614-623.

[16] Fan W M T. Metabolite profiling by one- and two-dimensional NMR analysis of complex mixtures[J]. Prog Nucl Magn Reson Spect, 1996, 28: 161-219.

[17] Nicholson J K, Foxall P J D, Spraul M, et al. 750 MHz ¹H and ¹H-¹³C NMR spectroscopy of human blood plasma[J]. Anal Chem, 1995, 67(5): 793-811.

[18] Puschel G P, Kirchner C, Schroder A, et al. Glycogenolytic and antiglycogenolytic prostaglandin E2 actions in rat hepatocytes are mediated via different signalling pathways[J]. Eur J Biochem, 1993, 218(3): 1 083-1 089.

[19] Mine T, Kojima I, Ogata E. Mechanism of prostaglandin E2-induced glucose production in rat hepatocytes[J]. Endocrinology, 1990, 126(6): 2 831-2 836.

[20] Puschel G P, Christ B. Inhibition by PGE2 of glucagon-induced increase in phosphoenolpyruvate carboxykinase mRNA and acceleration of mRNA degradation in cultured rat hepatocytes[J]. Febs Lett, 1994, 351(3): 353-356.

[21] Haussinger D, Stehle T, Tranthi T A, et al. Prostaglandin responses in isolated perfused rat liver: Ca²⁺ and K⁺ fluxes, hemodynamic and metabolic effects[J]. Biol Chem HoppeSeyler, 1987, 368(11): 1 509-1 513.

[22] Iwai M, Gardemann A, Puschel G, et al. Potential role for prostaglandin F2α, D2, E2 and thromboxane A2 in mediating the metabolic and hemodynamic actions of sympathetic nerves in perfused rat liver[J]. Eur J Biochem, 1988, 175(1): 45-50.

[23] Okumura T, Saito K. Effect of prostaglandins on glycogenesis and glycogenolysis in primary cultures of rathepatocytes: Arole of prostaglandin D2 in the liver[J]. Prostaglandins, 1990, 39(5): 525-540.

[24] Okumura T, Saito K. A sex difference in the effect of prostaglandins on hormone-stimulated glycogenolysis in primary cultures of rat hepatocytes[J]. Biochim Biophys Acta, 1990, 1 051(3): 300-305.

[25] Ames B N, Elson-Schwab I, Silver E A. Highdose vitamin therapy stimulates variant enzymes with decreased coenzyme binding affinity (increased K-m): relevance to genetic disease and polymorphisms[J]. Am J Clin Nutr, 2002, 75(4): 616-658.

[26] Kojima M, Morisaki T, Izuhara K, et al. Lipopolysaccharide increases cyclo-oxygenase-2 expression in a colon carcinoma cell line through nuclear factor-kappa B activation[J]. Oncogene, 2000, 19(9): 1 225-1 231.

[27] Johnson L R, Guthrie P. Effect of cholecystokinin and 16,16-dimethyl prostaglandin E2 on RNA and DNA of gastric and duodenal mucosa[J]. Gastroenterology, 1976, 70(1): 59-65.

[28] Smith K L, Tisdale M J. Mechanism of muscle protein degradation in cancer cachexia[J]. British J Cancer, 1993, 68(2): 314-318.

[29] Tisdale M J. Inhibition of lipolysis and muscle protein degradation by EPA in cancer cachexia[J]. Nutrition, 1996, 12(1): S31-S33.

[30] Kitada H, Miyata M, Nakamura T, et al. Protective role of hydroxysteroid sulfotransferase in lithocholic acid-induced liver toxicity[J]. J Biol Chem, 2003, 278(20): 17 838-17 844.

[31] Ljubuncic P, Tanne Z, Bomzon A. Evidence of a systemic phenomenon for oxidative stress in cholestatic liver disease[J]. Gut, 2000, 47(5): 710-716.

[32] Jaeschke H, Farhood A. Neutrophil and Kupffer cell-induced oxidant stress and ischemia-reperfusion injury in rat liver[J]. Am J Physiol, 1991, 260(3): G355-G362.

[33] Masubuchi Y, Suda C, Horie T. Involvement of mitochondrial permeability transition in acetaminophen-induced liver injury in mice[J]. J Hepatol, 2005, 42(1): 110-116.