塞络通胶囊治疗急性脑缺血模型大鼠的代谢组学分析
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摘要
目的:从代谢组学角度研究塞络通胶囊治疗急性多发性脑梗死大鼠模型的作用机制。方法:24只SD大鼠随机分为假手术组、模型组和塞络通组(33 mg·kg~(-1)),通过大鼠颈内动脉注射荧光微球的方法建立急性多发性脑梗死大鼠模型。手术成功后,塞络通组大鼠通过十二指肠注射给药,给药体积2 m L·kg~(-1)。基于UPLC-Q-TOF-MS分析各组大鼠脑组织中内源性代谢物,应用主成分分析(PCA)和偏最小二乘法-判别分析(PLS-DA)进行模式识别及生物标志物的鉴定。结果:通过模式识别发现假手术组与模型组的代谢轮廓有明显差异,并鉴定了10个与急性脑缺血相关的生物标志物。与假手术组相比,模型组中N-乙酰天门冬氨酸(NAA),延胡索酸,谷胱甘肽,脱氢抗坏血酸,天门冬氨酸以及S-腺苷同型半胱氨酸含量下降,精氨酸、瓜氨酸、酵母氨酸、乙内酰脲-5-丙酸含量升高;而塞络通胶囊则可使上述10个异常变化的生物标志物水平显著回调。结论:塞络通胶囊主要调节的代谢途径有NAA代谢、精氨酸代谢、能量代谢及氧化应激等。
Objective: To explore the mechanism of Sailuotong capsules in treating acute cerebral ischemia from the perspective of metabonomics. Method: A total of 24 SD rats were randomly divided into 3 groups,including sham-operated group,model group and Sailuotong group( 33 mg·kg-1). The rat model of acute multiple cerebral infarction was established by injecting fluorescent microspheres into internal carotid artery. After the successful operation,rats in Sailuotong group were administered by duodenal injection immediately,and the dosage volume was 2 m L·kg-1. Endogenous metabolites in rat brain tissues of each group were determined by UPLC-Q-TOF-MS. The relevant data and biomarkers were analyzed by principal component analysis( PCA) and partial least squares-discriminant analysis( PLS-DA). Result: The analysis of pattern recognition indicated that the metabolite profiles in model group and sham-operated group were separated obviously,and ten biomarkers related to acute cerebral ischemia were also identified. Compared with the sham-operated group,contents of N-acetylaspartate( NAA), fumaric acid, glutathione, dehydroascorbic acid, aspartic acid and Sadenosylhomocysteine were decreased,while the contents of arginine,citrulline,saccharopine and hydantoin-5-propionic acid were increased in the model group. Meanwhile,the ten abnormal biomarkers mentioned above got restoration in Sailuotong group. Conclusion: The main regulated metabolic pathways of Sailuotong capsules are NAA metabolism,arginine metabolism,energy metabolism,oxidative stress,etc.
Objective: To explore the mechanism of Sailuotong capsules in treating acute cerebral ischemia from the perspective of metabonomics. Method: A total of 24 SD rats were randomly divided into 3 groups,including sham-operated group,model group and Sailuotong group( 33 mg·kg-1). The rat model of acute multiple cerebral infarction was established by injecting fluorescent microspheres into internal carotid artery. After the successful operation,rats in Sailuotong group were administered by duodenal injection immediately,and the dosage volume was 2 m L·kg-1. Endogenous metabolites in rat brain tissues of each group were determined by UPLC-Q-TOF-MS. The relevant data and biomarkers were analyzed by principal component analysis( PCA) and partial least squares-discriminant analysis( PLS-DA). Result: The analysis of pattern recognition indicated that the metabolite profiles in model group and sham-operated group were separated obviously,and ten biomarkers related to acute cerebral ischemia were also identified. Compared with the sham-operated group,contents of N-acetylaspartate( NAA), fumaric acid, glutathione, dehydroascorbic acid, aspartic acid and Sadenosylhomocysteine were decreased,while the contents of arginine,citrulline,saccharopine and hydantoin-5-propionic acid were increased in the model group. Meanwhile,the ten abnormal biomarkers mentioned above got restoration in Sailuotong group. Conclusion: The main regulated metabolic pathways of Sailuotong capsules are NAA metabolism,arginine metabolism,energy metabolism,oxidative stress,etc.
引文
[1] YIN Y W,LI J C,LI B H,et al. Toll-like receptor 4 gene Asp299Gly polymorphism in ischemic cerebrovascular disease:a meta-analysis[J]. Int J Neurosci,2014,124(4):252-260.
[2]张静,赵建军,杨伟,等.基于中医证候特征的缺血性中风复发风险的横断面分析[J].中国实验方剂学杂志,2018,24(12):194-198.
[3] WANG H P,SHI S J,YAN W J,et al. Ischemic stroke susceptibility gene in a Northern Han Chinese population[J]. Neural Regen Res,2013,8(20):1881-1891.
[4]吴玉芙,刘晓红,郭伟成,等.补阳还五汤对气虚血瘀型脑梗死患者脑血管储备功能的影响[J].中国实验方剂学杂志,2017,23(12):162-167.
[5]张颖,林力,刘建勋,等.塞络通灌胃后人参皂苷在大鼠体内的药代动力学及脑分布研究[J].中国中药杂志,2014,39(2):316-321.
[6]张颖,林力,刘建勋,等.中药复方塞络通中银杏内酯类成分的药代动力学及脑分布研究[J].世界科学技术—中医药现代化,2014,16(7):1458-1464.
[7]徐立,宋文婷,林成仁,等.塞络通胶囊对大鼠多发性脑梗死恢复期Glu和GABA合成以及NMDA受体亚型表达的影响[J].药学学报,2012,47(7):870-877.
[8]徐立,丛伟红,魏翠娥,等.维脑康对小鼠记忆障碍的影响[J].中药药理与临床,2007,23(6):60-62.
[9]徐立,林成仁,刘建勋,等.塞络通胶囊对多发梗死性痴呆大鼠认知功能障碍的干预作用[J].中国中药杂志,2012,37(19):2943-2946.
[10]徐立,刘建勋,丛伟红,等.维脑康胶囊对慢性脑低灌注大鼠脑内胆碱能系统及清除自由基能力的影响[J].中国中药杂志,2008,33(5):531-534.
[11]任建勋,徐立,林成仁,等.塞络通胶囊在络脉瘀阻证脑梗死大鼠缺血时间窗外给药的保护作用[J].中华中医药杂志,2012,27(12):3060-3063.
[12]郭慧,崔扬,王秋红,等.基于代谢组学技术的中药复方研究近况[J].中国实验方剂学杂志,2017,23(1):213-219.
[13]孟胜喜,霍清萍,王兵,等.代谢组学技术在中医药防治缺血性脑血管病研究中的应用[J].中医杂志,2016,57(17):1515-1518,1521.
[14] Takagi N,Miyake K,Taguchi T,et al. Failure in learning task and loss of cortical cholinergic fibers in microsphere-embolized rats[J]. Exp Brain Res,1997,114(2):279-287.
[15] Koller K J,Coyle J T. Ontogenesis of N-acetyl-aspartate and N-acetyl-aspartyl-glutamate in rat brain[J]. Brain Res,1984,317(1):137-140.
[16]蔺俊斌,廖维靖. N-乙酰天冬氨酸与缺血性卒中的研究进展[J].中国脑血管病杂志,2011,8(9):501-504.
[17]刘雪辉,曹平,钟文,等.天智颗粒联合盐酸多奈哌齐片对轻中度肝阳上亢证血管性痴呆的临床分析[J].中国实验方剂学杂志,2016,22(7):177-181.
[18] Muoz Maniega S,Cvoro V,Chappell F M,et al. Changes in NAA and lactate following ischemic stoke:a serial MR spectroscopic imaging study[J]. Neurology,2008,71(24):1993-1999.
[19] Demougeot C, Marie C, Giroud M, et al. Nacetylaspartate:a literature review of animal research on brain ischaemia[J]. J Neurochem,2004,90(4):776-783.
[20] LEI H,Berthet C,Hirt L,et al. Evolution of the neurochemical profile after transient focal cerebral ischemia in the mouse brain[J]. J Cereb Blood Flow Metab,2009,29(4):811-819.
[21] Lemesle M,Walker P,Guy F,et al. Multi-variate analysis predicts clinical outcome 30 days after middle cerebral artery infarction[J]. Acta Neurol Scand,2000,102(1):11-17.
[22] van Zandvoort M J,van der Grond J,Kappelle L J,et al. Cognitive deficits and changes in neurometabolites after a lacunar infarct[J]. J Neurol,2005,252(2):183-190.
[23] Morris D,Khurasany M,Nguyen T,et al. Glutathione and infection[J]. Biochim Biophy Acta,2013,1830(5):3329-3349.
[24] Lyons J,Rauh-Pfeiffer A,YU Y M,et al. Blood glutathione synthesis rates in healthy adults receiving a sulfur amino acid-free diet[J]. Proc Natl Acad Sci USA,2000,97(10):5071-5076.
[25]易文龙.谷胱甘肽对新生大鼠脑缺氧缺血脑细胞内钙和脂质过氧化影响的研究[J].数理医药学杂志,2004,17(4):371-372.
[26]郭志丽,朱妍,肖红斌,等.与脑梗死关联的氨基酸代谢通路研究进展与发现[J].中国医药导报,2013,10(26):24-27.
[27]陈瑞群,廖成彬,郭茜,等.结合血清代谢组学与支持向量回归机揭示右归丸调节肾阳虚证的代谢标志物群及其作用机制[J].中国实验方剂学杂志,2018,24(13):62-70.
[28] Rector T S,Bank A J,Mullen K A,et al. Randomized,double-blind,placebo-controlled study of supplemental oral L-arginine in patients with heart failure[J].Circulation,1996,93(12):2135-2141.
[2]张静,赵建军,杨伟,等.基于中医证候特征的缺血性中风复发风险的横断面分析[J].中国实验方剂学杂志,2018,24(12):194-198.
[3] WANG H P,SHI S J,YAN W J,et al. Ischemic stroke susceptibility gene in a Northern Han Chinese population[J]. Neural Regen Res,2013,8(20):1881-1891.
[4]吴玉芙,刘晓红,郭伟成,等.补阳还五汤对气虚血瘀型脑梗死患者脑血管储备功能的影响[J].中国实验方剂学杂志,2017,23(12):162-167.
[5]张颖,林力,刘建勋,等.塞络通灌胃后人参皂苷在大鼠体内的药代动力学及脑分布研究[J].中国中药杂志,2014,39(2):316-321.
[6]张颖,林力,刘建勋,等.中药复方塞络通中银杏内酯类成分的药代动力学及脑分布研究[J].世界科学技术—中医药现代化,2014,16(7):1458-1464.
[7]徐立,宋文婷,林成仁,等.塞络通胶囊对大鼠多发性脑梗死恢复期Glu和GABA合成以及NMDA受体亚型表达的影响[J].药学学报,2012,47(7):870-877.
[8]徐立,丛伟红,魏翠娥,等.维脑康对小鼠记忆障碍的影响[J].中药药理与临床,2007,23(6):60-62.
[9]徐立,林成仁,刘建勋,等.塞络通胶囊对多发梗死性痴呆大鼠认知功能障碍的干预作用[J].中国中药杂志,2012,37(19):2943-2946.
[10]徐立,刘建勋,丛伟红,等.维脑康胶囊对慢性脑低灌注大鼠脑内胆碱能系统及清除自由基能力的影响[J].中国中药杂志,2008,33(5):531-534.
[11]任建勋,徐立,林成仁,等.塞络通胶囊在络脉瘀阻证脑梗死大鼠缺血时间窗外给药的保护作用[J].中华中医药杂志,2012,27(12):3060-3063.
[12]郭慧,崔扬,王秋红,等.基于代谢组学技术的中药复方研究近况[J].中国实验方剂学杂志,2017,23(1):213-219.
[13]孟胜喜,霍清萍,王兵,等.代谢组学技术在中医药防治缺血性脑血管病研究中的应用[J].中医杂志,2016,57(17):1515-1518,1521.
[14] Takagi N,Miyake K,Taguchi T,et al. Failure in learning task and loss of cortical cholinergic fibers in microsphere-embolized rats[J]. Exp Brain Res,1997,114(2):279-287.
[15] Koller K J,Coyle J T. Ontogenesis of N-acetyl-aspartate and N-acetyl-aspartyl-glutamate in rat brain[J]. Brain Res,1984,317(1):137-140.
[16]蔺俊斌,廖维靖. N-乙酰天冬氨酸与缺血性卒中的研究进展[J].中国脑血管病杂志,2011,8(9):501-504.
[17]刘雪辉,曹平,钟文,等.天智颗粒联合盐酸多奈哌齐片对轻中度肝阳上亢证血管性痴呆的临床分析[J].中国实验方剂学杂志,2016,22(7):177-181.
[18] Muoz Maniega S,Cvoro V,Chappell F M,et al. Changes in NAA and lactate following ischemic stoke:a serial MR spectroscopic imaging study[J]. Neurology,2008,71(24):1993-1999.
[19] Demougeot C, Marie C, Giroud M, et al. Nacetylaspartate:a literature review of animal research on brain ischaemia[J]. J Neurochem,2004,90(4):776-783.
[20] LEI H,Berthet C,Hirt L,et al. Evolution of the neurochemical profile after transient focal cerebral ischemia in the mouse brain[J]. J Cereb Blood Flow Metab,2009,29(4):811-819.
[21] Lemesle M,Walker P,Guy F,et al. Multi-variate analysis predicts clinical outcome 30 days after middle cerebral artery infarction[J]. Acta Neurol Scand,2000,102(1):11-17.
[22] van Zandvoort M J,van der Grond J,Kappelle L J,et al. Cognitive deficits and changes in neurometabolites after a lacunar infarct[J]. J Neurol,2005,252(2):183-190.
[23] Morris D,Khurasany M,Nguyen T,et al. Glutathione and infection[J]. Biochim Biophy Acta,2013,1830(5):3329-3349.
[24] Lyons J,Rauh-Pfeiffer A,YU Y M,et al. Blood glutathione synthesis rates in healthy adults receiving a sulfur amino acid-free diet[J]. Proc Natl Acad Sci USA,2000,97(10):5071-5076.
[25]易文龙.谷胱甘肽对新生大鼠脑缺氧缺血脑细胞内钙和脂质过氧化影响的研究[J].数理医药学杂志,2004,17(4):371-372.
[26]郭志丽,朱妍,肖红斌,等.与脑梗死关联的氨基酸代谢通路研究进展与发现[J].中国医药导报,2013,10(26):24-27.
[27]陈瑞群,廖成彬,郭茜,等.结合血清代谢组学与支持向量回归机揭示右归丸调节肾阳虚证的代谢标志物群及其作用机制[J].中国实验方剂学杂志,2018,24(13):62-70.
[28] Rector T S,Bank A J,Mullen K A,et al. Randomized,double-blind,placebo-controlled study of supplemental oral L-arginine in patients with heart failure[J].Circulation,1996,93(12):2135-2141.