CN113189346B - 一组检测生脉注射液质量的血清蛋白标志物及其应用 - Google Patents
一组检测生脉注射液质量的血清蛋白标志物及其应用 Download PDFInfo
- Publication number
- CN113189346B CN113189346B CN202110461175.2A CN202110461175A CN113189346B CN 113189346 B CN113189346 B CN 113189346B CN 202110461175 A CN202110461175 A CN 202110461175A CN 113189346 B CN113189346 B CN 113189346B
- Authority
- CN
- China
- Prior art keywords
- serum
- injection
- proteins
- group
- shengmai injection
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 238000002347 injection Methods 0.000 title claims abstract description 58
- 239000007924 injection Substances 0.000 title claims abstract description 58
- 239000009877 shengmai Substances 0.000 title claims abstract description 33
- 102000004506 Blood Proteins Human genes 0.000 title abstract description 9
- 108010017384 Blood Proteins Proteins 0.000 title abstract description 9
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 68
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 68
- 208000031225 myocardial ischemia Diseases 0.000 claims abstract description 22
- 230000003285 pharmacodynamic effect Effects 0.000 claims abstract description 17
- JWZZKOKVBUJMES-UHFFFAOYSA-N (+-)-Isoprenaline Chemical compound CC(C)NCC(O)C1=CC=C(O)C(O)=C1 JWZZKOKVBUJMES-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000000090 biomarker Substances 0.000 claims description 16
- 229940039009 isoproterenol Drugs 0.000 claims description 14
- 210000002966 serum Anatomy 0.000 abstract description 37
- 238000000034 method Methods 0.000 abstract description 20
- 238000011160 research Methods 0.000 abstract description 16
- 230000001105 regulatory effect Effects 0.000 abstract description 12
- 238000012216 screening Methods 0.000 abstract description 8
- 238000012795 verification Methods 0.000 abstract description 6
- 239000003550 marker Substances 0.000 abstract description 5
- 229940126680 traditional chinese medicines Drugs 0.000 abstract description 5
- 238000013441 quality evaluation Methods 0.000 abstract description 4
- 229960001317 isoprenaline Drugs 0.000 abstract 2
- 241000700159 Rattus Species 0.000 description 24
- 239000003814 drug Substances 0.000 description 17
- WEVYAHXRMPXWCK-UHFFFAOYSA-N Acetonitrile Chemical compound CC#N WEVYAHXRMPXWCK-UHFFFAOYSA-N 0.000 description 15
- 150000001875 compounds Chemical class 0.000 description 14
- 239000000243 solution Substances 0.000 description 12
- 230000000694 effects Effects 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 10
- 108090000765 processed proteins & peptides Proteins 0.000 description 9
- 238000001819 mass spectrum Methods 0.000 description 8
- 102000003855 L-lactate dehydrogenase Human genes 0.000 description 7
- 108700023483 L-lactate dehydrogenases Proteins 0.000 description 7
- 102000007079 Peptide Fragments Human genes 0.000 description 7
- 108010033276 Peptide Fragments Proteins 0.000 description 7
- 102000019197 Superoxide Dismutase Human genes 0.000 description 7
- 108010012715 Superoxide dismutase Proteins 0.000 description 7
- 239000003153 chemical reaction reagent Substances 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 238000005516 engineering process Methods 0.000 description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 6
- 230000002107 myocardial effect Effects 0.000 description 6
- 238000004885 tandem mass spectrometry Methods 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 102000006602 glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 5
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 5
- 150000002500 ions Chemical class 0.000 description 5
- 238000004949 mass spectrometry Methods 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 4
- 230000009471 action Effects 0.000 description 4
- 238000005194 fractionation Methods 0.000 description 4
- 241000411851 herbal medicine Species 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 239000007791 liquid phase Substances 0.000 description 4
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 230000004224 protection Effects 0.000 description 4
- 238000011002 quantification Methods 0.000 description 4
- 238000000108 ultra-filtration Methods 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- DTQVDTLACAAQTR-UHFFFAOYSA-N Trifluoroacetic acid Chemical compound OC(=O)C(F)(F)F DTQVDTLACAAQTR-UHFFFAOYSA-N 0.000 description 3
- 101710159648 Uncharacterized protein Proteins 0.000 description 3
- 230000033228 biological regulation Effects 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- 239000012634 fragment Substances 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 239000006166 lysate Substances 0.000 description 3
- 238000012544 monitoring process Methods 0.000 description 3
- 239000012071 phase Substances 0.000 description 3
- 239000002504 physiological saline solution Substances 0.000 description 3
- 102000004196 processed proteins & peptides Human genes 0.000 description 3
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 2
- 102100037819 Fas apoptotic inhibitory molecule 1 Human genes 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 101000878509 Homo sapiens Fas apoptotic inhibitory molecule 1 Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 102000004142 Trypsin Human genes 0.000 description 2
- 108090000631 Trypsin Proteins 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 230000006907 apoptotic process Effects 0.000 description 2
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 2
- 239000004202 carbamide Substances 0.000 description 2
- 230000005961 cardioprotection Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000003828 downregulation Effects 0.000 description 2
- 238000010828 elution Methods 0.000 description 2
- 235000019253 formic acid Nutrition 0.000 description 2
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 230000002401 inhibitory effect Effects 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 238000002955 isolation Methods 0.000 description 2
- 238000002372 labelling Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 238000004811 liquid chromatography Methods 0.000 description 2
- 238000011068 loading method Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 239000012188 paraffin wax Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000012588 trypsin Substances 0.000 description 2
- 238000002211 ultraviolet spectrum Methods 0.000 description 2
- 230000003827 upregulation Effects 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 229940088872 Apoptosis inhibitor Drugs 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102000004420 Creatine Kinase Human genes 0.000 description 1
- 108010042126 Creatine kinase Proteins 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 108090000604 Hydrolases Proteins 0.000 description 1
- 102000004157 Hydrolases Human genes 0.000 description 1
- 108010044467 Isoenzymes Proteins 0.000 description 1
- 239000007836 KH2PO4 Substances 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 241000699666 Mus <mouse, genus> Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 240000002948 Ophiopogon intermedius Species 0.000 description 1
- 235000002789 Panax ginseng Nutrition 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 239000006004 Quartz sand Substances 0.000 description 1
- 240000006079 Schisandra chinensis Species 0.000 description 1
- 235000008422 Schisandra chinensis Nutrition 0.000 description 1
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 101710100179 UMP-CMP kinase Proteins 0.000 description 1
- 102000054569 UMP-CMP kinases Human genes 0.000 description 1
- XBJFCYDKBDVADW-UHFFFAOYSA-N acetonitrile;formic acid Chemical compound CC#N.OC=O XBJFCYDKBDVADW-UHFFFAOYSA-N 0.000 description 1
- 235000012538 ammonium bicarbonate Nutrition 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000000259 anti-tumor effect Effects 0.000 description 1
- 239000000158 apoptosis inhibitor Substances 0.000 description 1
- 238000003149 assay kit Methods 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- -1 carbamoylmethyl Chemical group 0.000 description 1
- 206010007625 cardiogenic shock Diseases 0.000 description 1
- 230000003293 cardioprotective effect Effects 0.000 description 1
- 239000000919 ceramic Substances 0.000 description 1
- 238000013375 chromatographic separation Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000010494 dissociation reaction Methods 0.000 description 1
- 230000005593 dissociations Effects 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- 230000008030 elimination Effects 0.000 description 1
- 238000003379 elimination reaction Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000007717 exclusion Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- HQVFCQRVQFYGRJ-UHFFFAOYSA-N formic acid;hydrate Chemical compound O.OC=O HQVFCQRVQFYGRJ-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- PGLTVOMIXTUURA-UHFFFAOYSA-N iodoacetamide Chemical compound NC(=O)CI PGLTVOMIXTUURA-UHFFFAOYSA-N 0.000 description 1
- 238000005468 ion implantation Methods 0.000 description 1
- 238000001294 liquid chromatography-tandem mass spectrometry Methods 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 208000010125 myocardial infarction Diseases 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000036961 partial effect Effects 0.000 description 1
- 238000010827 pathological analysis Methods 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000000575 proteomic method Methods 0.000 description 1
- 238000003908 quality control method Methods 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 238000004366 reverse phase liquid chromatography Methods 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000000870 ultraviolet spectroscopy Methods 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 210000001631 vena cava inferior Anatomy 0.000 description 1
- 230000002861 ventricular Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
- G01N33/6803—General methods of protein analysis not limited to specific proteins or families of proteins
- G01N33/6848—Methods of protein analysis involving mass spectrometry
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N30/00—Investigating or analysing materials by separation into components using adsorption, absorption or similar phenomena or using ion-exchange, e.g. chromatography or field flow fractionation
- G01N30/02—Column chromatography
- G01N30/88—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86
- G01N2030/8809—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample
- G01N2030/8813—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials
- G01N2030/8831—Integrated analysis systems specially adapted therefor, not covered by a single one of the groups G01N30/04 - G01N30/86 analysis specially adapted for the sample biological materials involving peptides or proteins
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N2800/00—Detection or diagnosis of diseases
- G01N2800/32—Cardiovascular disorders
- G01N2800/325—Heart failure or cardiac arrest, e.g. cardiomyopathy, congestive heart failure
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Immunology (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Hematology (AREA)
- General Health & Medical Sciences (AREA)
- Biomedical Technology (AREA)
- Analytical Chemistry (AREA)
- Urology & Nephrology (AREA)
- General Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Pathology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biotechnology (AREA)
- Food Science & Technology (AREA)
- Bioinformatics & Computational Biology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Cell Biology (AREA)
- Biophysics (AREA)
- Microbiology (AREA)
- Organic Chemistry (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Water Supply & Treatment (AREA)
- Genetics & Genomics (AREA)
- Other Investigation Or Analysis Of Materials By Electrical Means (AREA)
Abstract
本发明公开了一组检测生脉注射液质量的血清蛋白标志物及其应用,属于中药领域。本发明利用血清蛋白质组学方法,筛选得到了一组受异丙肾上腺素(ISO)诱导的心肌缺血调控,同时生脉注射液干预对其具有回调作用的蛋白,并选择了其中的三个蛋白进行了PRM验证,结果也验证了所述蛋白质作为生脉注射液质量标志物的可靠性。本发明还提供了一种有效的用于生脉注射液质量评价的血清标志物筛选方法,此方法能够有效的筛选血清中生脉注射液的标志物,为进一步筛选更多生脉注射液药效学指标奠定了基础。本发明公开的标志物同时能够作为心肌缺血的诊断标志物,并且在生脉注射液质量评价和药效学研究中具有广泛的应用价值。
Description
技术领域
本发明涉及中药领域,特别是涉及一组检测生脉注射液质量的血清蛋白标志物及其应用。
背景技术
中药复方注射剂在临床上应用广泛、作用确切,但是其安全性一直是备受关注的问题。体内过程研究包括药代动力学、药效学以及药动学-药效学结合研究有助于指导中药复方的临床合理用药,对于保障中药复方的有效性和安全性具有重要意义。目前,随着分析技术的进步,中药复方药代动力学研究已经取得了很大进展。但是,中药药效学由于缺乏明确、易获取的药效指标,其研究仍然面临许多问题和挑战。对于复方制剂来说,药物的组分配比是影响药物效果的关键。目前在复方药物的配比优化研究中,常常通过给药后的整体治疗效果来评价药物配方,但其存在检测指标过多、精确度较低的问题。
生脉注射液是由红参、麦冬、五味子三味中药组成的复方制剂,在临床上广泛用于治疗心肌梗塞、心源性休克、感染性休克等症。目前对于生脉注射液质量和药效标志物的研究多集中于心肌组织,但其样本的获取困难,不能做到实时监控,且取样时需要处死小鼠,增加大量实验成本。
血清蛋白质组学(serum proteomics)是生命科学研究领域一门新兴而发展迅速的学科,其采取系统的、全方位的研究模式,从整体、动态、网络的水平上对与疾病相关和药物作用下的所有血清蛋白质进行研究。血液中含有丰富的蛋白质,特别是包含了来源于许多机体组织的蛋白质子集,血清蛋白质组学研究可以提供比研究单一组织或器官更加广泛的蛋白质种类和数量。因此,血清蛋白质组学技术已经成为研究抗肿瘤以及抗心血管系统疾病机制及靶点筛选的有力工具。但目前尚未有通过血清蛋白质组学研究生脉注射液质量鉴定标志物的相关报道。
发明内容
本发明的目的是提供一组检测生脉注射液质量的血清蛋白标志物及其应用,以解决上述现有技术存在的问题,为生脉注射液的质量控制和药效学研究提供有用的工具。
为实现上述目的,本发明提供了如下方案:
技术方案之一,一组检测生脉注射液质量的心肌缺血生物标志物,所述的生物标志物包括ProteinID分别为D4AA05、Q4KM73、M0R5J4、M0R590、Q5M871或D3ZJW6的蛋白质中的一种或多种。
在一些优选的技术方案中,所述的生物标志物包括ProteinID分别为M0R590、Q5M871、M0R5J4的蛋白质中的一种或多种。
技术方案之二,提供了检测所述的生物标志物的试剂在制备诊断心肌缺血的制剂中的应用。
技术方案之三,提供了检测所述的生物标志物的试剂在生脉注射液药效学研究中的应用。
技术方案之四,提供了检测所述的生物标志物的试剂在制备评价生脉注射液质量的试剂中的应用。
技术方案之五,提供了一种检测试剂盒,所述的试剂盒中包括检测所述的生物标志物的试剂。
技术方案之六,提供了一种生脉注射液质量评价血清生物标志物的筛选方法,所述方法包括以下步骤:
(1)样品提取:取生脉组注射生脉注射液的心肌缺血大鼠、模型组未注射生脉注射液的心肌缺血大鼠和对照组普通大鼠血清样本,去除高丰度蛋白质,得到低丰度组分溶液;超滤浓缩,加入裂解液,离心,取上清;采用BCA法进行蛋白质定量;
(2)TMT标记:各血清样品消化后,对照组、模型组和生脉组各三个生物学重复样本分别以126、127N,127C,128N,128C,129N,129C,130N和130C标记;每组标记后的肽段等量混合,采用High pH RP spin column进行分级,分级样品冻干后用12μL 0.1%FA复溶,在280nm的UV光谱下测定肽段浓度;
(3)色谱分离与质谱分析:采用液相色谱分离样品,分离后的样品进行质谱分析,搜索质谱数据,得到差异蛋白。
在一些优选的实施方案中,所述的液相色谱条件为:缓冲液A液为0.1%甲酸水溶液,B液为0.1%甲酸乙腈水溶液,其中乙腈为84%;色谱柱以95%的A液平衡,流速为300nL/min;分级后的肽段样品分别进行1.5小时梯度洗脱,液相梯度为:0min-80min,B液线性梯度从0%-55%;80min-85min,B液线性梯度从55%-100%;85min-90min,B液维持在100%。
在一些优选的实施方案中,所述的质谱分析条件为:检测方式为正离子,母离子扫描范围300–1800m/z,一级质谱质荷比为200m/z,分辨率为70,000,AGC靶点为3e6,一级最大注射时间为10ms,扫描范围数为1,动态排除时间为40.0s;每次全扫描后采集10个碎片图谱MS2 scan,MS2激活类型为HCD,隔离窗为2m/z,二级质谱质荷比为200m/z,分辨率为35,000,Microscans为1,二级最大注射时间为60ms,归一化碰撞能量为30eV,Underfill为0.1%,质谱分析90分钟。
本发明公开了以下技术效果:
本发明利用血清蛋白质组学方法,筛选得到了一组受ISO诱导的心肌缺血调控,同时生脉注射液干预对其具有回调作用的蛋白,并选择了其中的三个蛋白进行了PRM验证,结果也显示了准确性。本发明还提供了一种有效的生脉注射液质量评价血清标志物的筛选方法,此方法能够有效的筛选血清中生脉注射液的标志物,为进一步筛选更多生脉注射液药效学指标奠定了基础。本发明公开的标志物同时能够作为心肌缺血的诊断标志物,并且在生脉注射液质量评价和药效学研究中具有广泛的应用价值。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的一些实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据这些附图获得其他的附图。
图1为本发明TMT方法的流程图;
图2为生脉注射液对ISO诱导的大鼠心肌缺血损伤的心脏保护作用图;图中A为对照组、模型组、生脉高剂量组和生脉低剂量组大鼠的心肌组织H&E图(200×);B为对照组、模型组、生脉高剂量组和生脉低剂量组大鼠血清LDH,CK-MB和SOD水平(Mean±SD,n=10),##P<0.01vs.对照组,*P<0.05vs.模型组);
图3为所有差异表达蛋白质韦恩图;
图4为TMT和PRM获得的三个血清差异蛋白M0R5J4,M0R590和Q5M871的平均表达量变化倍数比较图(Mean±SD,n=3)。
具体实施方式
现详细说明本发明的多种示例性实施方式,该详细说明不应认为是对本发明的限制,而应理解为是对本发明的某些方面、特性和实施方案的更详细的描述。
应理解本发明中所述的术语仅仅是为描述特别的实施方式,并非用于限制本发明。另外,对于本发明中的数值范围,应理解为还具体公开了该范围的上限和下限之间的每个中间值。在任何陈述值或陈述范围内的中间值以及任何其他陈述值或在所述范围内的中间值之间的每个较小的范围也包括在本发明内。这些较小范围的上限和下限可独立地包括或排除在范围内。
除非另有说明,否则本文使用的所有技术和科学术语具有本发明所述领域的常规技术人员通常理解的相同含义。虽然本发明仅描述了优选的方法和材料,但是在本发明的实施或测试中也可以使用与本文所述相似或等同的任何方法和材料。本说明书中提到的所有文献通过引用并入,用以公开和描述与所述文献相关的方法和/或材料。在与任何并入的文献冲突时,以本说明书的内容为准。
在不背离本发明的范围或精神的情况下,可对本发明说明书的具体实施方式做多种改进和变化,这对本领域技术人员而言是显而易见的。由本发明的说明书得到的其他实施方式对技术人员而言是显而易见得的。本申请说明书和实施例仅是示例性的。
实施例1
1、材料与方法
1.1材料与试剂
生脉注射液(生产批号:16060604)购自由江苏苏中药业集团股份有限公司。异丙肾上腺素(Isoproterenol,ISO)(sigma公司,美国);乳酸脱氢酶(LDH)、肌酸激酶同工酶MB(CK-MB)测定试剂盒(南京建成生物工程研究所);总超氧化物歧化酶(SOD)活性检测试剂盒(南京建成生物工程研究所)。
甘油(G0854生工/500ml)、溴酚蓝(161-0404生工)、SDS(161-0302Bio-Rad)、Urea(161-0731Bio-Rad)、Tris(A6141 Sigma)、二硫苏糖醇(DTT,161-0404Bio-Rad)、碘乙酰胺(IAA,163-2109Bio-Rad)、KH2PO4(10017618国药)、KCl(10016318国药)、HCl(10011018国药)、BCA定量试剂盒(P0012碧云天)、BSA(A0332-25G生工)、NH4HCO3(A6141 Sigma)、Trypsin(317107Promega)、甲酸(FA,06450Fluka)、三氟乙酸(TFA,T6508 Sigma)、乙腈(ACN,I592230123Merck)、10kD超滤离心管(Sartorius)、石英砂(MP 6910-050)、1/4英寸陶瓷珠(MP 6540-034)、TMT 6/10plex Isobaric Label Reagent(Thermo)
Multiple Affinity Removal LC Column–Human 14/Mouse 3(Agilent)、PierceTMHigh pH Reversed-Phase Peptide Fractionation Kit;C18上样柱:Thermo ScientificAcclaim PepMap100,100μm*2cm,nanoViper C18,3μm,C18分析柱:Thermoscientific EASY column,10cm,ID75μm,3μm,C18-A2;5X上样缓冲液:10%SDS,0.5%溴酚蓝,50%甘油,500mM DTT,250mM TrisHCl,pH6.8;SDT裂解液:4%SDS,100mM Tris-HCl,1mMDTT,pH7.6;UA buffer:8M urea,150mM Tris-HCl,pH 8.0;HPLC流动相A:0.1%FA;HPLC流动相B:0.1%FA,84%ACN
1.2实验动物
成年健康SD大鼠30只,雄性,体质量220~240g,由浙江省实验动物中心提供。动物许可证号:SCXK(浙)2014-0001。动物实验遵照嘉兴学院动物伦理审查委员会批准的动物保护和使用原则开展。
1.3实验分组及给药
30只大鼠随机分为对照组、模型组、生脉注射液组,每组10只。根据生脉注射液临床常用剂量折算大鼠给药剂量,为10.8mL/kg/天,ISO造模前尾静脉(iv)给药,连续给药7天。对照组和模型组分别尾静脉(iv)给予相应体积的生理盐水。模型组、生脉注射液组分别于给药后两天(第8、9天)按85mg/kg/d剂量连续皮下注射ISO生理盐水溶液制造急性心肌缺血模型,对照组皮下注射相应体积的生理盐水。
1.4样品采集与检测
每组大鼠于末次给药ISO(或生理盐水)后30min,开胸进行下腔静脉取血,室温放置1h后,取血清;同时摘取大鼠心脏,生理盐水洗去残血。取左心室部分用福尔马林固定,脱水石蜡包埋,常规切片,用于HE染色。
大鼠血清分成若干份(约300μL/份),-80℃保存。根据试剂盒方法分别测定各组血清中的LDH、SOD、CK-MB等生化指标。采用石蜡包埋、HE染色方法进行心肌组织的病理学分析。
1.5 TMT血清蛋白质组学分析
选取3个组别的血清样本用于TMT蛋白质组学研究,3个组别分别为:对照组,模型组和生脉注射液组。每组样品分别各取3份等量混匀,组成1个生物学重复样本,每组含有3个生物学重复样本,共计9个样本。总体流程如图1所示。
1.5.1血清高丰度蛋白的去除及蛋白定量
取适量血清样本采用去血清高丰度亲和色谱柱Agilent Multiple AffinityRemoval LC Column-Mouse 3按照Agilent对应protocol中的操作方法去除高丰度蛋白质,得到低丰度组分溶液。应用10kD超滤管进行超滤浓缩,加入一倍体积的SDT裂解液,沸水浴15min,14000g离心20min,取上清。采用BCA法进行蛋白质定量。分装样品,-80℃保存。
1.5.2 TMT标记和分级
使用过滤器辅助的样品前处理方法消化各血清样品,得到的肽段在280nm的UV光谱下进行定量。各样品分别取100μg肽段,按照Thermo公司TMT标记试剂盒说明书进行标记,本研究中对照组、模型组和生脉组各三个生物学重复样本分别以126、127N,127C,128N,128C,129N,129C,130N和130C标记。将每组标记后的肽段等量混合,采用High pH RP spincolumn进行分级。分级样品冻干后用12μL 0.1%FA复溶,在280nm的UV光谱下测定肽段浓度。
1.5.3 LC-MS/MS分析
每份样品采用纳升流速的HPLC液相系统Easy nLC进行分离。缓冲液A液为0.1%甲酸水溶液,B液为0.1%甲酸乙腈水溶液(乙腈为84%)。色谱柱以95%的A液平衡,样品由自动进样器上样到上样柱(Thermo Scientific Acclaim PepMap100,100μm*2cm,nanoViperC18),经过分析柱(Thermo scientific EASY column,10cm,ID75μm,3μm,C18-A2)分离,流速为300nL/min。分级后的肽段样品分别进行1.5小时梯度洗脱,液相梯度为:0min-80min,B液线性梯度从0%-55%;80min-85min,B液线性梯度从55%-100%;85min-90min,B液维持在100%。
样品经色谱分离后用Q-Exactive质谱仪进行质谱分析。分析时长为90min,检测方式为正离子,母离子扫描范围300–1800m/z,一级质谱(200m/z)分辨率为70,000,AGC靶点为3e6,一级最大注射时间为10ms,扫描范围数为1,动态排除时间为40.0s。每次全扫描(fullscan)后采集10个碎片图谱(MS2 scan),MS2激活类型为HCD,隔离窗为2m/z,二级质谱(200m/z)分辨率为35,000,Microscans为1,二级最大注射时间为60ms,归一化碰撞能量为30eV,Underfill为0.1%。
1.5.4蛋白鉴定和定量分析
采用Proteome Discoverer 1.4软件(Thermo Scientifc)中的MASCOT引擎(Matrix Science,伦敦,英国)搜索MS/MS质谱数据。设置参数为:所有胰蛋白酶具特异性;氨基甲酰甲基(C),TMT 10plex(N-末端)和TMT10plex(赖氨酸,K)设置为固定修饰,氧化(蛋氨酸,M)和TMT 10plex(酪氨酸,Y)设置为可变修饰,肽对于所有采集的MS1质谱图,质量公差设置为20ppm,对于所有MS2质谱图的碎片,质量公差设置为0.1Da。错误发现率(FDR)设置为≤0.01。蛋白质比率以蛋白质唯一肽的中值计算。所有肽比率均通过中位数蛋白质比率进行标准化,标准化后中位数蛋白质比率应为“1”。差异蛋白上调阈值设置为比较组的比率>1.2且P值<0.05,下调阈值设置为比较组的比率<0.83且P值<0.05。
1.6 PRM验证
为了验证TMT分析获得的蛋白质表达水平,采用平行反应监视(parallelreaction monitoring,PRM)技术对目标差异蛋白进行分析。根据TMT方案制备肽,并将AQUA稳定同位素肽掺入每个样品中作为内标参比。采用Easy nLC-1200系统(ThermoScientific)进行反相色谱分析,在45分钟内乙腈比例从5%到35%,液相分析时长为1小时。采用Q Exactive Plus质谱仪(Thermo Scientific)进行PRM分析:检测方式为正离子,一级质谱MS1全扫描(200m/z)分辨率为70,000,AGC靶点为3e6,最大离子注入时间250ms;每次一级MS1扫描后根据Inclustion list采集20个PRM扫描(MS2扫描),MS2扫描(200m/z)分辨率为35,000,AGC靶点为3e6,最大注射时间200ms,Microscans为1,解离窗为2Th,归一化碰撞能为27。采用Skyline(MacCoss实验室,华盛顿大学)分析原始数据。
1.7数据处理
所有数据用均数±标准差(Mean±SD)表示,统计学采用SPSS 16.0软件进行组间学生t检验分析,P<0.05认为具有显著性差异。
2实验结果
2.1生脉注射液抗大鼠心肌缺血的心脏保护效应
通过分析生脉注射液对心肌缺血大鼠的心肌组织病理变化及对血清LDH,CK-MB和SOD水平的影响,综合评价其心脏保护效应。如图2(A)所示,与模型组大鼠相比,高、低剂量生脉注射液预处理均可适度减轻大鼠心肌炎症细胞的浸润以及纤维化现象。如图2(B)所示,与对照组相比,模型组大鼠血清LDH、CK-MB水平显著升高,SOD水平显著降低(P<0.01);与模型组相比,生脉注射液高、低剂量组能显著降低血清LDH、CK-MB水平,升高血清SOD水平(P<0.05)
2.2血清差异表达蛋白筛选
实验结果共鉴定到蛋白质1009个,其中558个蛋白含有至少两个唯一肽段。在三个比较组分别定量获得1000个蛋白,以倍数变化大于1.2倍(上调大于1.2倍或者下调小于0.83)且P<0.05的标准筛选差异表达蛋白质,三个比较组模型组/对照组、生脉组/模型组和生脉组/对照组分别得到差异表达蛋白227(85上调,142下调)、100(14上调,86下调)和228(115上调,113下调)个(表1)。表2为生脉组/模型组的差异表达蛋白信息,表3为模型组/对照组的差异表达蛋白信息。组间差异蛋白的重叠情况参见韦恩图(图3),研究发现在所有差异蛋白中,与ISO诱导大鼠心肌缺血和生脉注射液调控作用同时相关的共有10个血清蛋白(表4)。
表1.TMT蛋白质组学鉴定获得的组间血清差异蛋白数量
表2.生脉组/模型组大鼠血清显著上调和下调的差异蛋白
表3.模型组/对照组大鼠血清显著上调和下调的差异蛋白
表4.生脉组/模型组和模型组/对照组中的10个共同差异表达蛋白
2.3 PRM验证
采用PRM对TMT蛋白质组学技术获得的部分血清差异蛋白进行验证,选取差异蛋白的标准是:(1)与ISO诱导大鼠心肌缺血和生脉注射液调控作用都相关的共同血清差异蛋白;(2)蛋白在ISO诱导的心肌缺血大鼠血清中具有差异表达,同时生脉注射液可反调该蛋白的表达;(3)由TMT技术获得的唯一肽段大于1的血清蛋白。依据以上标准,本研究选取了三个差异蛋白M0R5J4,M0R590和Q5M871用于PRM验证,如图4所示,PRM和TMT获得的三个差异蛋白的组间相对表达量变化的趋势相似,说明本研究TMT技术获得的血清蛋白质组学数据具可靠性。
3、结果分析
中药复方注射剂在临床上应用广泛、作用确切,但是其安全性一直是备受关注的问题。体内过程研究包括药代动力学、药效学以及药动学-药效学结合研究有助于指导中药复方的临床合理用药,对于保障中药复方的有效性和安全性具有重要意义。目前,随着分析技术的进步,中药复方药代动力学研究已经取得了很大进展。但是,中药药效学由于缺乏明确、易获取的药效指标,其研究仍然面临许多问题和挑战。血清蛋白质组学不仅可以帮助系统地揭示药物作用的靶标和机制,而且有助于筛选潜在的生物标志物,为中药复方的药代动力学和药效学研究提供有价值的候选血清药效指标。本研究基于血清蛋白质组学技术,发现了与生脉注射液调控作用相关的100个血清差异蛋白,筛选到的蛋白数量和种类远远多于心肌组织蛋白质组学。根据韦恩图分析,本发明在模型组/对照组和生脉组/模型组中发现了10个共同的差异蛋白(表3),其中有6个蛋白受ISO诱导的心肌缺血调控,同时生脉注射液干预对其具有回调作用,它们分别是羧酸酯水解酶(ProteinID-D4AA05)、UMP-CMP激酶(ProteinID-Q4KM73)、未表征蛋白质(ProteinID-M0R5J4)、甘油醛-3-磷酸脱氢酶(ProteinID-M0R590)、Fas凋亡抑制分子3(ProteinID-Q5M871)和RCG21066(ProteinID-D3ZJW6)。这6个蛋白显然应该成为生脉注射液心脏保护作用的潜在生物标志物,我们对其中的甘油醛-3-磷酸脱氢酶(GAPDH)、Fas细胞凋亡抑制分子3(FAIM3)和一个未表征的蛋白质(M0R5J4)进行了PRM验证,结果证明其组间表达量的变化趋势与TMT蛋白质学的研究结果相似。
本发明采用TMT定量蛋白质组学技术对生脉注射液抗ISO诱导大鼠心肌缺血损伤的心脏保护作用进行了血清蛋白质组学分析。研究结果分别在模型组/对照组、生脉组/模型组和生脉组/对照组三个比较组中各鉴定获得227、100和228个血清差异蛋白,血清蛋白质组学筛选获得的差异蛋白数量和种类远多于心肌组织蛋白质组学。其中有6个蛋白受ISO诱导的心肌缺血调控,同时生脉注射液干预对其具有回调作用,选择了三个蛋白:甘油醛-3-磷酸脱氢酶(GAPDH)、Fas细胞凋亡抑制分子3(FAIM3)和一个未表征的蛋白质(M0R5J4)进行了PRM验证,结果证明了TMT蛋白质组学的准确性,这些血清蛋白可作为评价生脉注射液质量及开展药效学研究的生物标志物。另外,本发明还获得了模型组与对照组之间差异表达的227个蛋白,这些蛋白可以作为心肌缺血的潜在生物标志物。
以上所述的实施例仅是对本发明的优选方式进行描述,并非对本发明的范围进行限定,在不脱离本发明设计精神的前提下,本领域普通技术人员对本发明的技术方案做出的各种变形和改进,均应落入本发明权利要求书确定的保护范围内。
Claims (3)
1.一种检测生脉注射液质量的心肌缺血生物标志物,所述的生物标志物包括ProteinID分别为M0R5J4、M0R590、Q5M871的蛋白质中的一种或多种;
所述心肌缺血为异丙肾上腺素诱导的心肌缺血。
2.权利要求1所述的生物标志物在生脉注射液药效学研究中的应用。
3.权利要求1所述的生物标志物在评价生脉注射液质量中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110461175.2A CN113189346B (zh) | 2021-04-27 | 2021-04-27 | 一组检测生脉注射液质量的血清蛋白标志物及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110461175.2A CN113189346B (zh) | 2021-04-27 | 2021-04-27 | 一组检测生脉注射液质量的血清蛋白标志物及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113189346A CN113189346A (zh) | 2021-07-30 |
CN113189346B true CN113189346B (zh) | 2024-03-22 |
Family
ID=76979545
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110461175.2A Active CN113189346B (zh) | 2021-04-27 | 2021-04-27 | 一组检测生脉注射液质量的血清蛋白标志物及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113189346B (zh) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN117007810B (zh) * | 2023-06-12 | 2024-04-05 | 中国医学科学院北京协和医院 | 用于预测静脉内平滑肌瘤进展的生物标志物及其应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101363837A (zh) * | 2008-09-02 | 2009-02-11 | 浙江大学 | 一种胰腺癌化疗药疗效评价方法 |
CN105891314A (zh) * | 2016-03-09 | 2016-08-24 | 上海中科新生命生物科技有限公司 | 一种iTRAQ技术筛选心肌梗死疾病表达蛋白的方法 |
CN109115933A (zh) * | 2018-08-31 | 2019-01-01 | 中国人民解放军第四军医大学 | 一种筛选四逆汤抗心肌缺血药效物质基础的方法 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US20100121360A9 (en) * | 2000-12-20 | 2010-05-13 | Fox Hollow Technologies, Inc | Testing a patient population having a cardiovascular condition for drug efficacy |
US7022478B2 (en) * | 2002-05-14 | 2006-04-04 | The Chinese University Of Hong Kong | Methods for evaluating stroke or cardiac ischemia by nucleic acid detection |
US20090269796A1 (en) * | 2007-12-05 | 2009-10-29 | The General Hospital Corporation | Methods of detecting and treating myocardial ischemia and myocardial infarction |
WO2012058313A2 (en) * | 2010-10-27 | 2012-05-03 | The Broad Institute Of Mit And Harvard | Novel biomarkers for cardiovascular injury |
US20140147874A1 (en) * | 2011-03-04 | 2014-05-29 | The Johns Hopkins University | Biomarkers of cardiac ischemia |
-
2021
- 2021-04-27 CN CN202110461175.2A patent/CN113189346B/zh active Active
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101363837A (zh) * | 2008-09-02 | 2009-02-11 | 浙江大学 | 一种胰腺癌化疗药疗效评价方法 |
CN105891314A (zh) * | 2016-03-09 | 2016-08-24 | 上海中科新生命生物科技有限公司 | 一种iTRAQ技术筛选心肌梗死疾病表达蛋白的方法 |
CN109115933A (zh) * | 2018-08-31 | 2019-01-01 | 中国人民解放军第四军医大学 | 一种筛选四逆汤抗心肌缺血药效物质基础的方法 |
Non-Patent Citations (4)
Title |
---|
"Discovery of potential plasma protein biomarkers for acute myocardial infarction via proteomics";Xu Shasha等;《Journal of Thoracic Disease》;第11卷(第9期);第3962-3972页 * |
"A Metabolomic Study of Rats with Doxorubicin-Induced Cardiomyopathy and Shengmai Injection Treatment";Chen Yu等;《PLOS ONE》;第10卷(第5期);第1-20页 * |
"Quantitative Proteomics for Cardiac Biomarker Discovery Using Isoproterenol-Treated Nonhuman Primates";Song Benben等;《Journal of Proteome》;第13卷;第5909-5917页 * |
"基于串联质谱标签法和平行反应监测技术的氟喹诺酮耐药沙门菌蛋白质组学分析";李琳等;《微生物学通报》;第45卷(第7期);第1535-1545页 * |
Also Published As
Publication number | Publication date |
---|---|
CN113189346A (zh) | 2021-07-30 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Marshall et al. | Beyond the paradigm: Combining mass spectrometry and nuclear magnetic resonance for metabolomics | |
Courant et al. | Development of a metabolomic approach based on liquid chromatography-high resolution mass spectrometry to screen for clenbuterol abuse in calves | |
CN103616450B (zh) | 一种肺癌患者血清特异性代谢产物谱及其建立方法 | |
US7653496B2 (en) | Feature selection in mass spectral data | |
CN106248838B (zh) | 高通量液相色谱串联质谱的检测方法以及检测4种儿茶酚胺代谢物的方法 | |
CN115015460B (zh) | 一种运用广泛靶向代谢组学技术鉴定冬虫夏草产地的方法 | |
CN107037146B (zh) | 一种复方木鸡颗粒质量控制的方法 | |
Santacruz et al. | Metabolomic study of soft corals from the Colombian Caribbean: PSYCHE and 1H-NMR comparative analysis | |
Li et al. | Rapid characterization of chemical markers for discrimination of Moutan Cortex and its processed products by direct injection-based mass spectrometry profiling and metabolomic method | |
CN113189346B (zh) | 一组检测生脉注射液质量的血清蛋白标志物及其应用 | |
Zhang et al. | Quality evaluation of traditional Chinese drug toad venom from different origins through a simultaneous determination of bufogenins and indole alkaloids by HPLC | |
CN112986441A (zh) | 一种从组织代谢轮廓筛选的肿瘤标志物及其应用和辅助诊断方法 | |
Musharraf et al. | Metabolite profiling of human plasma by different extraction methods through gas chromatography–mass spectrometry—An objective comparison | |
Abul-Husn et al. | Neuroproteomics of the synapse and drug addiction | |
Liu et al. | Metabolomic study of a rat fever model induced with 2, 4-dinitrophenol and the therapeutic effects of a crude drug derived from Coptis chinensis | |
CN114624317B (zh) | 一种基于直接进样质谱的定性和定量分析方法 | |
CN107907610A (zh) | 用于急性过敏反应豚鼠的血清代谢生物标志物 | |
Liu et al. | Simultaneous determination of eight tryptic peptides in musk using high-performance liquid chromatography coupled with tandem mass spectrometry | |
Callipo et al. | Evaluation of different two‐dimensional chromatographic techniques for proteomic analysis of mouse cardiac tissue | |
Shyur et al. | Metabolomics in herbal medicine research | |
KR101444579B1 (ko) | 인삼 연근 판별용 바이오마커 조성물 | |
CN112630344B (zh) | 代谢标志物在脑梗死中的用途 | |
CN111948326B (zh) | 一种桑黄指纹图谱的构建方法、指纹图谱及应用 | |
Liu et al. | LC/MS fingerprint and simultaneous quantification of main bioactive compounds in Polyporus umbellatus (Pers.) Fr. from different regions and developmental stages | |
Luo et al. | Comparison of liquid chromatography-high-resolution tandem mass spectrometry (MS2) and multi-stage mass spectrometry (MS3) for screening toxic natural products |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |