CN106692123B - Application of gamma-aminobutyric acid in preparation of heart protection pharmaceutical preparation - Google Patents
Application of gamma-aminobutyric acid in preparation of heart protection pharmaceutical preparation Download PDFInfo
- Publication number
- CN106692123B CN106692123B CN201611203181.3A CN201611203181A CN106692123B CN 106692123 B CN106692123 B CN 106692123B CN 201611203181 A CN201611203181 A CN 201611203181A CN 106692123 B CN106692123 B CN 106692123B
- Authority
- CN
- China
- Prior art keywords
- gaba
- heart
- free
- fermentation liquor
- solution
- 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
- BTCSSZJGUNDROE-UHFFFAOYSA-N gamma-aminobutyric acid Chemical compound NCCCC(O)=O BTCSSZJGUNDROE-UHFFFAOYSA-N 0.000 title claims abstract description 234
- OGNSCSPNOLGXSM-UHFFFAOYSA-N (+/-)-DABA Natural products NCCC(N)C(O)=O OGNSCSPNOLGXSM-UHFFFAOYSA-N 0.000 title claims abstract description 111
- 229960003692 gamma aminobutyric acid Drugs 0.000 title claims abstract description 111
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 230000004224 protection Effects 0.000 title abstract description 14
- 239000000825 pharmaceutical preparation Substances 0.000 title description 6
- 238000000855 fermentation Methods 0.000 claims abstract description 59
- 230000004151 fermentation Effects 0.000 claims abstract description 59
- 230000000694 effects Effects 0.000 claims abstract description 18
- 210000005003 heart tissue Anatomy 0.000 claims abstract description 18
- 108010025020 Nerve Growth Factor Proteins 0.000 claims abstract description 15
- 210000002064 heart cell Anatomy 0.000 claims abstract description 15
- 102000015779 HDL Lipoproteins Human genes 0.000 claims abstract description 14
- 108010010234 HDL Lipoproteins Proteins 0.000 claims abstract description 14
- 244000005700 microbiome Species 0.000 claims abstract description 13
- 239000002699 waste material Substances 0.000 claims abstract description 13
- 241000251468 Actinopterygii Species 0.000 claims abstract description 11
- 102000008214 Glutamate decarboxylase Human genes 0.000 claims abstract description 8
- 108091022930 Glutamate decarboxylase Proteins 0.000 claims abstract description 8
- 238000012545 processing Methods 0.000 claims abstract description 8
- 239000002994 raw material Substances 0.000 claims abstract description 5
- 239000000243 solution Substances 0.000 claims description 46
- 239000007788 liquid Substances 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 claims description 17
- 239000011347 resin Substances 0.000 claims description 15
- 229920005989 resin Polymers 0.000 claims description 15
- 239000000725 suspension Substances 0.000 claims description 15
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 claims description 14
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 14
- WSMYVTOQOOLQHP-UHFFFAOYSA-N Malondialdehyde Chemical compound O=CCC=O WSMYVTOQOOLQHP-UHFFFAOYSA-N 0.000 claims description 14
- 230000001580 bacterial effect Effects 0.000 claims description 14
- 229940118019 malondialdehyde Drugs 0.000 claims description 14
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 12
- 102000007072 Nerve Growth Factors Human genes 0.000 claims description 11
- 239000003900 neurotrophic factor Substances 0.000 claims description 11
- 229940073490 sodium glutamate Drugs 0.000 claims description 11
- 238000001179 sorption measurement Methods 0.000 claims description 10
- PXEDJBXQKAGXNJ-QTNFYWBSSA-L disodium L-glutamate Chemical compound [Na+].[Na+].[O-]C(=O)[C@@H](N)CCC([O-])=O PXEDJBXQKAGXNJ-QTNFYWBSSA-L 0.000 claims description 9
- 238000004128 high performance liquid chromatography Methods 0.000 claims description 9
- 235000013923 monosodium glutamate Nutrition 0.000 claims description 9
- 230000017531 blood circulation Effects 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 8
- 230000002829 reductive effect Effects 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 7
- VHUUQVKOLVNVRT-UHFFFAOYSA-N Ammonium hydroxide Chemical compound [NH4+].[OH-] VHUUQVKOLVNVRT-UHFFFAOYSA-N 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 239000012670 alkaline solution Substances 0.000 claims description 6
- 235000011114 ammonium hydroxide Nutrition 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims description 5
- 230000033228 biological regulation Effects 0.000 claims description 5
- 230000003683 cardiac damage Effects 0.000 claims description 5
- 238000004042 decolorization Methods 0.000 claims description 5
- 238000005238 degreasing Methods 0.000 claims description 5
- 239000008367 deionised water Substances 0.000 claims description 5
- 229910021641 deionized water Inorganic materials 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 5
- 238000010828 elution Methods 0.000 claims description 5
- 239000001257 hydrogen Substances 0.000 claims description 5
- 229910052739 hydrogen Inorganic materials 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 210000001519 tissue Anatomy 0.000 claims description 5
- 229910052799 carbon Inorganic materials 0.000 claims description 4
- 238000001914 filtration Methods 0.000 claims description 4
- 238000005406 washing Methods 0.000 claims description 4
- 210000003403 autonomic nervous system Anatomy 0.000 claims description 3
- 238000000227 grinding Methods 0.000 claims description 3
- 230000008569 process Effects 0.000 claims description 3
- 238000010009 beating Methods 0.000 claims description 2
- 230000015572 biosynthetic process Effects 0.000 claims description 2
- 210000004165 myocardium Anatomy 0.000 claims description 2
- 230000001737 promoting effect Effects 0.000 claims description 2
- 238000003786 synthesis reaction Methods 0.000 claims description 2
- 238000003828 vacuum filtration Methods 0.000 claims description 2
- KRHYYFGTRYWZRS-UHFFFAOYSA-M Fluoride anion Chemical compound [F-] KRHYYFGTRYWZRS-UHFFFAOYSA-M 0.000 claims 1
- 230000006698 induction Effects 0.000 claims 1
- 210000004027 cell Anatomy 0.000 abstract description 9
- 210000004369 blood Anatomy 0.000 abstract description 7
- 239000008280 blood Substances 0.000 abstract description 7
- 230000002107 myocardial effect Effects 0.000 abstract description 6
- 102000015336 Nerve Growth Factor Human genes 0.000 abstract description 4
- 229940053128 nerve growth factor Drugs 0.000 abstract description 4
- 230000032683 aging Effects 0.000 abstract description 3
- 235000016709 nutrition Nutrition 0.000 abstract description 3
- 230000035764 nutrition Effects 0.000 abstract description 3
- 230000003647 oxidation Effects 0.000 abstract description 3
- 238000007254 oxidation reaction Methods 0.000 abstract description 3
- 230000006907 apoptotic process Effects 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 18
- 230000008859 change Effects 0.000 description 14
- 239000013642 negative control Substances 0.000 description 12
- 239000013641 positive control Substances 0.000 description 9
- 239000008213 purified water Substances 0.000 description 9
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- 230000003203 everyday effect Effects 0.000 description 8
- 235000019688 fish Nutrition 0.000 description 8
- PUZPDOWCWNUUKD-UHFFFAOYSA-M sodium fluoride Chemical compound [F-].[Na+] PUZPDOWCWNUUKD-UHFFFAOYSA-M 0.000 description 8
- 230000000747 cardiac effect Effects 0.000 description 7
- 238000001514 detection method Methods 0.000 description 7
- 230000006870 function Effects 0.000 description 7
- 230000002354 daily effect Effects 0.000 description 6
- ACTIUHUUMQJHFO-UHFFFAOYSA-N Coenzym Q10 Natural products COC1=C(OC)C(=O)C(CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UHFFFAOYSA-N 0.000 description 5
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 5
- 235000017471 coenzyme Q10 Nutrition 0.000 description 5
- 229940110767 coenzyme Q10 Drugs 0.000 description 5
- ACTIUHUUMQJHFO-UPTCCGCDSA-N coenzyme Q10 Chemical compound COC1=C(OC)C(=O)C(C\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UPTCCGCDSA-N 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 230000000877 morphologic effect Effects 0.000 description 5
- 238000010172 mouse model Methods 0.000 description 5
- 241001468179 Enterococcus avium Species 0.000 description 4
- 240000001929 Lactobacillus brevis Species 0.000 description 4
- 235000013957 Lactobacillus brevis Nutrition 0.000 description 4
- 244000057717 Streptococcus lactis Species 0.000 description 4
- 235000014897 Streptococcus lactis Nutrition 0.000 description 4
- 229940024606 amino acid Drugs 0.000 description 4
- 235000001014 amino acid Nutrition 0.000 description 4
- 150000001413 amino acids Chemical class 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 238000011161 development Methods 0.000 description 4
- 230000018109 developmental process Effects 0.000 description 4
- 235000013305 food Nutrition 0.000 description 4
- 210000000056 organ Anatomy 0.000 description 4
- 235000013024 sodium fluoride Nutrition 0.000 description 4
- 239000011775 sodium fluoride Substances 0.000 description 4
- 210000001835 viscera Anatomy 0.000 description 4
- 241001454694 Clupeiformes Species 0.000 description 3
- 241000282414 Homo sapiens Species 0.000 description 3
- 241000221523 Rhodotorula toruloides Species 0.000 description 3
- 238000000944 Soxhlet extraction Methods 0.000 description 3
- 235000019513 anchovy Nutrition 0.000 description 3
- 210000000601 blood cell Anatomy 0.000 description 3
- 210000004204 blood vessel Anatomy 0.000 description 3
- 210000000170 cell membrane Anatomy 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 238000011981 development test Methods 0.000 description 3
- 239000004519 grease Substances 0.000 description 3
- 238000009629 microbiological culture Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 230000001681 protective effect Effects 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- 230000003068 static effect Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000011282 treatment Methods 0.000 description 3
- 201000001320 Atherosclerosis Diseases 0.000 description 2
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
- 239000004471 Glycine Substances 0.000 description 2
- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 description 2
- 102000004895 Lipoproteins Human genes 0.000 description 2
- 108090001030 Lipoproteins Proteins 0.000 description 2
- 240000007994 Rhodomyrtus tomentosa Species 0.000 description 2
- 235000007234 Rhodomyrtus tomentosa Nutrition 0.000 description 2
- 241000269851 Sarda sarda Species 0.000 description 2
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 2
- 235000004279 alanine Nutrition 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 210000003855 cell nucleus Anatomy 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 208000029078 coronary artery disease Diseases 0.000 description 2
- 239000003085 diluting agent Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 230000003859 lipid peroxidation Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 230000002503 metabolic effect Effects 0.000 description 2
- 210000002569 neuron Anatomy 0.000 description 2
- 235000018102 proteins Nutrition 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 239000002516 radical scavenger Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000035882 stress Effects 0.000 description 2
- 238000000967 suction filtration Methods 0.000 description 2
- OIXLLKLZKCBCPS-RZVRUWJTSA-N (2s)-2-azanyl-5-[bis(azanyl)methylideneamino]pentanoic acid Chemical compound OC(=O)[C@@H](N)CCCNC(N)=N.OC(=O)[C@@H](N)CCCNC(N)=N OIXLLKLZKCBCPS-RZVRUWJTSA-N 0.000 description 1
- 208000019901 Anxiety disease Diseases 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 206010003210 Arteriosclerosis Diseases 0.000 description 1
- 241000238424 Crustacea Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000146989 Engraulis japonicus Species 0.000 description 1
- PXGOKWXKJXAPGV-UHFFFAOYSA-N Fluorine Chemical compound FF PXGOKWXKJXAPGV-UHFFFAOYSA-N 0.000 description 1
- 208000013875 Heart injury Diseases 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241000269821 Scombridae Species 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 241000347391 Umbrina cirrosa Species 0.000 description 1
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 description 1
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000036506 anxiety Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 208000011775 arteriosclerosis disease Diseases 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 1
- 230000036983 biotransformation Effects 0.000 description 1
- 230000036772 blood pressure Effects 0.000 description 1
- 230000036760 body temperature Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000001569 carbon dioxide Substances 0.000 description 1
- 229910002092 carbon dioxide Inorganic materials 0.000 description 1
- 210000004413 cardiac myocyte Anatomy 0.000 description 1
- 230000005961 cardioprotection Effects 0.000 description 1
- 230000003293 cardioprotective effect Effects 0.000 description 1
- 239000007795 chemical reaction product Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 230000004087 circulation Effects 0.000 description 1
- 230000008602 contraction Effects 0.000 description 1
- 230000007123 defense Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 230000002124 endocrine Effects 0.000 description 1
- -1 fluoride ions Chemical class 0.000 description 1
- 229910052731 fluorine Inorganic materials 0.000 description 1
- 239000011737 fluorine Substances 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000004217 heart function Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 244000000013 helminth Species 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 235000020640 mackerel Nutrition 0.000 description 1
- 238000009364 mariculture Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 210000002464 muscle smooth vascular Anatomy 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 238000001694 spray drying Methods 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 231100000027 toxicology Toxicity 0.000 description 1
- 229940116269 uric acid Drugs 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/195—Carboxylic acids, e.g. valproic acid having an amino group
- A61K31/197—Carboxylic acids, e.g. valproic acid having an amino group the amino and the carboxyl groups being attached to the same acyclic carbon chain, e.g. gamma-aminobutyric acid [GABA], beta-alanine, epsilon-aminocaproic acid or pantothenic acid
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P13/00—Preparation of nitrogen-containing organic compounds
- C12P13/005—Amino acids other than alpha- or beta amino acids, e.g. gamma amino acids
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- General Health & Medical Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Public Health (AREA)
- Biotechnology (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Medicinal Chemistry (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention belongs to the technical field of biomedicine, and particularly relates to application of gamma-aminobutyric acid (GABA) in preparing a heart protection medicinal preparation. Fermenting low-value fish or processing waste thereof serving as a raw material by using a microorganism with glutamate decarboxylase activity, and separating and purifying the obtained fermentation product to obtain GABA. The experimental results show that: (1) GABA has obvious nutrition and protection effects on cardiac cells; (2) the oxidation resistance of GABA can inhibit the apoptosis of heart cells and prevent the aging of myocardial cells; (3) GABA can increase the content of high-density lipoprotein in heart blood; (4) GABA can increase the nerve growth factor content of heart tissue.
Description
Technical Field
The invention relates to the technical field of biomedicine, in particular to application of gamma-aminobutyric acid in preparation of a heart protection pharmaceutical preparation.
Background
The marine edible fish has rich nutrition, delicious taste, and many health functions, and has important edible and economic values. With the development of modern separation technology and biotechnology, the utilization of fish resources by people is no longer limited to the mariculture industry, and the research content relates to a plurality of fields of ecology, cultivation, medicine, toxicology, heredity, molecular biology and the like. The method carries out high-valued development and utilization on low-value fish resources, provides ideal marine food and marine medicine for human beings, creates macroscopic economic benefit, and is beneficial to promoting the rapid development of marine industry in China.
The heart is one of the most important organs in the vertebrate body. The heart functions to promote blood flow, provide sufficient blood flow to organs and tissues, supply oxygen and various nutrients (such as water, inorganic salts, glucose, proteins, various water-soluble vitamins, etc.), and remove metabolic end products (such as carbon dioxide, urea, uric acid, etc.), so that cells maintain normal metabolism and functions. Various endocrine hormones and other body fluid factors in the body are also transported to target cells through blood circulation to realize body fluid regulation and maintain the internal environment of the body relatively constant. In addition, the implementation of blood defense functions, and the regulation of body temperature that is relatively constant, also relies on the continuous circulation of blood within the blood vessels, which is achieved by the action of the heart "pump". The key of protecting heart is to strengthen the antioxidation performance of heart cells, promote blood circulation and reduce the cholesterol content in blood.
Gamma-aminobutyric acid (GABA) is an important inhibitory neurotransmitter in the nervous system of mammals, crustaceans, insects and certain parasitic worms. A few plants contain such non-protein amino acids. GABA is involved in various metabolic activities and has the effects of resisting oxidation, lowering blood pressure, improving sleep, resisting anxiety, improving lipid metabolism, reducing weight, preventing arteriosclerosis and the like, so that the GABA is paid more and more attention by scientific workers.
Disclosure of Invention
The invention aims to provide application of gamma-aminobutyric acid in preparation of a heart protection pharmaceutical preparation.
The scheme for achieving the purpose of the invention is as follows:
use of gamma-aminobutyric acid (GABA) for the preparation of a cardioprotective pharmaceutical preparation.
Preferably; application of gamma-aminobutyric acid (GABA) in preparing medicine for protecting heart damage caused by fluorine.
Preferably; the preparation method of GABA comprises the following steps: fermenting low-value fish or processing waste thereof serving as a raw material by using microorganisms with glutamate decarboxylase activity, and separating and purifying the obtained fermentation product to obtain the feed.
The method comprises the following steps:
1) grinding raw material low-value fish or processing waste thereof into homogenate, adding distilled water with 5-10 times volume (w/v) of homogenate mass into the homogenate to prepare homogenate, adding 0-5 wt% glucose (v/w) of the volume of the homogenate into the homogenate, and adjusting the pH value of the solution to 4.5-6.0 to obtain a microorganism culture solution. Sterilizing the culture solution, cooling, inoculating a microorganism with glutamate decarboxylase activity, and culturing at 30-40 ℃ for 24-72 hours to obtain a bacterial suspension (solution A) rich in the microorganism with the glutamate decarboxylase activity;
2) adjusting the pH value of the solution A to 5.5-6.5, adding 0.5-2 wt% of sodium glutamate (v/w) of the volume of the solution A to prepare a culture solution (solution B) rich in sodium glutamate, and culturing the solution B at 35-40 ℃ for 72-120 hours to obtain a fermentation liquid rich in GABA;
3) and (3) removing thallus and degreasing of GABA-rich fermentation liquor: centrifuging the fermentation liquid obtained in the step 2) for 5-20 minutes at the speed of 2000-;
4) and (3) decoloring thallus-free and grease-free fermentation liquor: decolorizing with active carbon or SD300 resin to obtain colorless transparent fermentation liquid;
5) and (3) separating and purifying GABA in colorless and transparent fermentation liquor: fully performing exchange adsorption on 732 resin (hydrogen type) exchange columns pretreated on the colorless and transparent fermentation liquor obtained in the step 4) at the flow rate of 2-5mL/min, washing the column to pH6.0 by deionized water after adsorption, washing the column by 100-200mL of 0.1-0.2mol/L alkaline solution, and then eluting the column by 1-2mol/L alkaline solution at the flow rate of 2-5 mL/min; and (3) developing with ninhydrin acetone solution during the elution process, collecting effluent with ninhydrin reaction showing blue color and pH of about 6.0, detecting GABA content in each tube of effluent by high performance liquid chromatography, combining the effluent with high GABA content, and concentrating under reduced pressure to obtain separated and purified GABA.
Preferably; the step 4) of adopting the activated carbon to decolor the thallus-free and grease-free fermentation liquor is to add 0.5 to 5 percent (w/v) of the activated carbon into the thallus-free and grease-free fermentation liquor, carry out water bath for 10 to 30 minutes at a temperature of between 60 and 90 ℃, and carry out vacuum filtration when the liquor is hot to obtain colorless and transparent fermentation liquor;
the SD300 resin decolorization is to decolorize thallus-free and grease-free fermentation liquor by adding 5-10% SD300 resin into the thallus-free and grease-free fermentation liquor, decolorizing at 20-30 ℃ for 3 hours by using a shaking table at 100rpm, and filtering to obtain colorless transparent fermentation liquor.
Preferably; and 5) concentrating under reduced pressure to 1/20-1/10 volume to obtain separated and purified GABA concentrated solution, and detecting the purity of GABA in the concentrated solution by using high performance liquid chromatography.
Preferably; and 5) the alkaline solution is a sodium hydroxide solution or ammonia water.
The heart damage model is fed with GABA (gamma-aminobutyric acid) which is separated and purified and has the concentration of 0.1-2.0g/L and is fed to a heart damage model mouse which is made of sodium fluoride for 5-20 days, and the GABA has obvious heart protection effect through the change of the morphological structure of heart cells, the malondialdehyde level of heart tissues and the change conditions of high-density lipoprotein and neurotrophic factor in heart blood.
Detecting the heart protection capability of the isolated and purified GABA:
1) a heart-damaged mouse model was established with sodium fluoride.
2) Feeding the separated and purified GABA with the concentration of 0.1-2.0g/L for 5-20 days.
3) The change conditions of the cell morphological structure of the heart cells, the malondialdehyde of the heart tissue, the high-density lipoprotein and the level of the neurotrophic factor show that the GABA has obvious heart protection effect.
The invention has the advantages that:
1) the twelfth order food grade GABA production process of Ministry of health in 2009: the L-sodium glutamate is used as a substrate and is produced by the steps of safe strain fermentation, heating sterilization, cooling, active carbon treatment, filtration, spray drying and the like. The present production line generally produces food grade GABA in batches with 20% purity, and GABA obtained through separation and purification of the present invention is prepared through biotransformation of L-sodium glutamate with safe strain, and has purity over 60% and purity higher than that of food grade GABA sold in the market.
2) The present invention provides a new use of GABA, the protection effect of GABA to heart can be realized from several aspects: (1) GABA has obvious nutrition and protection effects on cardiac cells; (2) the oxidation resistance of GABA can inhibit the apoptosis of heart cells and prevent the aging of myocardial cells; (3) GABA can increase the content of high density lipoprotein in heart blood. The high-density lipoprotein is plasma lipoprotein for resisting atherosclerosis, and is a protective factor for coronary heart disease. Commonly known as "blood vessel scavenger", it plays a key role in maintaining the normal function of the heart. (4) GABA can increase the nerve growth factor content of heart tissue. The movements of the vascular smooth muscle and the cardiac muscle in the heart are regulated by the autonomic nervous system. The nerve growth factor has important regulation and control functions on the development, differentiation, growth and regeneration of neurons distributed in the heart. By nourishing nerve cells in the heart, the blood flow in the heart is sufficient, and the myocardial contraction function can be normally exerted.
3) The GABA obtained by the invention is separated and purified from low-value fish or processing waste fermentation liquor thereof, and has the obvious characteristics of no toxicity and easy absorption.
Drawings
FIG. 1 is a microscopic structure diagram of cardiac cells under a 40-fold objective lens of a microscope, wherein (a) cardiac microstructures of a normal group, (b) cardiac microstructures of a negative control group, (c) cardiac microstructures of a positive control group, and (d) cardiac microstructures of a GABA group are isolated and purified;
FIG. 2 shows the detection of the purity of GABA separated and purified by high performance liquid chromatography.
Detailed Description
The following examples are further illustrative of the present invention and are not intended to be limiting thereof.
Example 1
1. Grinding fresh red bonito into homogenate by a grinder, adding distilled water with the mass of 10 volume (w/v) of the homogenate of the red bonito to prepare homogenate, adding glucose (v/w) with the volume of 1 wt% of the homogenate into the homogenate, adjusting the pH value of the solution to 5.5, and preparing a microorganism culture medium. The medium was sterilized at 121 ℃ for 15 minutes. After cooling, inoculating streptococcus lactis according to the inoculation amount (v/v) of 1 percent, and fermenting and culturing for 1 day at the temperature of 30 ℃ to obtain bacterial suspension rich in streptococcus lactis.
2. Adjusting the pH value of the bacterial suspension rich in the streptococcus lactis to 6.0, adding 1 wt% of sodium glutamate (v/w) of the volume of the bacterial suspension into the bacterial suspension, placing the bacterial suspension in a thermostat at 37 ℃, and performing static culture for 72 hours to obtain the fermented liquor of the rhodotorula gracilis with the GABA content of 4.8 g/L.
The streptococcus lactis is purchased from China general microbiological culture collection center, Beijing, and is numbered bio-52538.
3. And (3) separating and purifying GABA in the fermented liquor of the rhodomyrtus tomentosa rich in GABA.
1) And (3) removing thalli and degreasing of the fermented liquor of the rhodotorula gracilis rich in GABA: and (3) centrifuging the fermentation liquor obtained in the step (2) at the speed of 4000r/min for 10 minutes, taking the upper layer liquid, and carrying out autoclaving to obtain the sterile fermentation liquor. And removing grease from the sterile fermentation liquor by a Soxhlet extraction method to obtain thallus-free and grease-free fermentation liquor.
2) And (3) decoloring thallus-free and grease-free fermentation liquor: and (3) carrying out decolorization treatment by using SD300 resin, specifically adding 5% SD300 resin (w/v) into the obtained thallus-free and grease-free fermentation liquor, decolorizing for 3 hours at 25 ℃ by using a shaking table at 100rpm, and carrying out suction filtration to obtain colorless transparent fermentation liquor.
3) And (3) separating and purifying GABA in colorless and transparent fermentation liquor: the colorless transparent liquid was put on a 732 resin (hydrogen type) pretreated exchange column and sufficiently subjected to exchange adsorption at a flow rate of 3 mL/min. After adsorption, the mixture is washed by deionized water until the pH value is 6.0. The column was washed with 150mL of 0.1mol/L NaOH and then eluted with 1mol/L NaOH at a flow rate of 3 mL/min. The color development test was carried out with ninhydrin acetone solution continuously during the elution, and the change in pH was detected with pH paper. When the ninhydrin reaction is blue, and the pH value is about 6.0, retaining the effluent liquid until the ninhydrin blue reaction disappears, detecting the GABA content in the collection tube by high performance liquid chromatography, combining the effluent liquid in the collection tube with higher GABA content (the pH value of GABA aqueous solution is about 6, and the effluent liquid reacts with ninhydrin to be blue, so that the pH value of the effluent liquid can be judged to be 6, the effluent liquid reacts with ninhydrin to be blue, the amino acid contained in the effluent liquid is GABA), and concentrating under reduced pressure to 1/20 volumes to obtain the isolated and purified GABA concentrated solution.
Example 2
1. Adding 5 times (w/v) volume of distilled water into viscera (waste product) of the red-spotted maigre to prepare solution, adding 2.0 wt% of glucose (v/w) of the solution volume into the solution, adjusting pH value to 5.0, and preparing into microorganism culture solution. The culture broth was sterilized at 121 ℃ for 20 minutes. Cooling, inoculating enterococcus avium according to 1% inoculum size (v/v), fermenting and culturing at 37 deg.C for 36 hr to obtain suspension rich in enterococcus avium.
2. Adding 2 wt% of sodium glutamate (v/w) of the volume of the bacterial suspension into the bacterial suspension rich in the enterococcus avium, and adjusting the pH value to 5.5 to prepare a sodium glutamate-rich culture solution. The culture solution is placed in a thermostat with the temperature of 37 ℃ for static culture for 96 hours, and the fermented liquid of the internal organs of the red-brown mackerel with the GABA content of 5.7g/L is obtained.
The enterococcus avium is purchased from China general microbiological culture Collection center, Beijing, and the serial number is CGMCC 9184.
3. And (3) separating and purifying GABA in the fermented liquor of the viscera of the rhodomyrtus tomentosa rich in GABA.
1) And (3) removing thallus and degreasing from the fermented liquor of the internal organs of the rhodotorula gracilis rich in GABA: centrifuging at 3000r/min for 15 min, and sterilizing the upper layer liquid at high temperature and high pressure to obtain the sterile fermentation liquid. And removing grease from the thallus-free fermentation liquor by a Soxhlet extraction method to obtain the thallus-free and grease-free fermentation liquor.
2) And (3) decoloring thallus-free and grease-free fermentation liquor: and (3) carrying out decolorization treatment on SD300 resin, specifically adding 10% SD300 resin (w/v) into the thallus-free and grease-free fermentation liquor obtained above, decolorizing for 4 hours at 25 ℃ by using a shaking table at 100rpm, and carrying out suction filtration to obtain colorless transparent fermentation liquor.
3) And (3) separating and purifying GABA in colorless and transparent fermentation liquor: the colorless transparent liquid was passed through a 732 resin (hydrogen type) column which had been pretreated, and the flow rate was 3mL/min, and the column was sufficiently subjected to exchange adsorption. After adsorption, the mixture is washed by deionized water until the pH value is 6.0. The column was washed with 150mL of 0.1mol/L ammonia water, and then eluted with 1mol/L ammonia water at a flow rate of 3 mL/min. The color development test was carried out with ninhydrin acetone solution continuously during the elution, and the change in pH was detected with pH paper. When the ninhydrin reaction is blue, and the pH value is about 6.0, retaining the effluent, detecting the GABA content in the collecting tube by high performance liquid chromatography, combining the effluents in the collecting tube with higher GABA content (the pH value of the GABA aqueous solution is about 6, and the pH value of the GABA aqueous solution reacts with the ninhydrin to be blue, so that the effluent can be judged to have the pH value of 6, and reacts with the ninhydrin to be blue, and the effluent contains GABA, and concentrating under reduced pressure to 1/10 volume to obtain the separated and purified GABA concentrated solution.
Example 3
1. Decocting Engraulis japonicus Temminck et Schlegel in water, concentrating the waste liquid, adding 5 times (v/v) of distilled water to obtain diluent, adding 1 wt% of glucose (v/w) of the diluent, adjusting pH to 6.0, and making into microorganism culture solution. The culture broth was sterilized at 121 ℃ for 20 minutes. After cooling, inoculating lactobacillus brevis according to the inoculation amount (v/v) of 1 percent, and fermenting and culturing for 24 hours at the temperature of 37 ℃ to obtain bacterial suspension rich in lactobacillus brevis.
2. Adding 1.5 wt% of sodium glutamate (v/w) of the volume of the bacterial suspension into the bacterial suspension rich in the lactobacillus brevis, and adjusting the pH value to be 6.5 to prepare a sodium glutamate-rich culture solution. The culture solution is placed in a thermostat with the temperature of 37 ℃ for static culture for 100 hours, and anchovy poaching waste liquid fermentation liquor with the GABA content of 5.81g/L is obtained.
The lactobacillus brevis is purchased from China general microbiological culture collection center, Beijing, and has the number of CGMCC 1.2028.
3. And (3) separating and purifying GABA in the anchovy water-boiling waste liquid fermentation liquor rich in GABA.
1) Removing thallus and degreasing from the anchovy poaching waste liquid fermentation liquor rich in GABA: centrifuging at 5000r/min for 10 min, and sterilizing the upper layer liquid at high temperature and high pressure to obtain sterile fermentation liquid. And removing grease from the thallus-free fermentation liquor by a Soxhlet extraction method to obtain the thallus-free and grease-free fermentation liquor.
2) Activated carbon decolorization of thallus-free and grease-free fermentation liquor: adding 1.5% active carbon (w/v) into the thallus-free and grease-free fermentation liquor, decoloring for 30 minutes at 80 ℃, and filtering to obtain colorless transparent fermentation liquor.
3) And (3) separating and purifying GABA in colorless and transparent fermentation liquor: the colorless transparent fermentation liquid is put on a 732 resin (hydrogen type) pretreated exchange column, and the flow rate is 3mL/min, so that the colorless transparent fermentation liquid is fully subjected to exchange adsorption. After adsorption, the mixture is washed by deionized water until the pH value is 6.0. The column was washed with 150mL of 0.1mol/L ammonia water, and then eluted with 1mol/L ammonia water at a flow rate of 3 mL/min. The color development test was carried out with ninhydrin acetone solution continuously during the elution, and the change in pH was detected with pH paper. When the ninhydrin reaction is blue, and pH is about 6.0, retaining the effluent, detecting GABA content in the collection tube by high performance liquid chromatography, and combining the effluents in the collection tube with higher GABA content (the pH of GABA aqueous solution is about 6, and reacts with ninhydrin to be blue, so that the pH of the effluent can be judged to be 6, and the effluent reacts with ninhydrin to be blue, and contains GABA). The GABA concentrated solution is separated and purified by concentrating under reduced pressure to 1/10 volumes.
The GABA concentrated solutions separated and purified in the above examples 1, 2 and 3 were combined, and the purity of GABA was measured by HPLC, and the results are shown in FIG. 2, in which the GABA content was 63%, and the other detectable amino acids were: ala alanine 2%, Gly glycine 8%, Arg arginine 5%. Alanine, glycine and arginine are all common amino acids in human bodies, have low content, are all in a safe intake range, and cannot generate adverse physiological influence on human bodies or tested animals.
Example 4
The combined GABA concentrated solution obtained by separation and purification is used for preparing the heart protection pharmaceutical preparation, and the test and the effect of the heart protection effect are as follows:
cardioprotection test of isolated and purified GABA:
experimental animals: 18-22 g/male mouse of 120 Kunming species.
Animal model: 100 mice were fed 0.1g/L NaF for 30 days, creating heart-damaged model mice.
Preparing the medicine: the GABA concentrated solutions separated and purified in the above examples 1, 2 and 3 were combined, and then purified water was added to the combined solution to prepare a solution having GABA contents of 0.1g/L, 0.5g/L and 1.0 g/L.
Grouping experiments: 20 normal groups (healthy mice were fed purified water daily); 20 negative control groups (heart-impaired mice were fed purified water daily); 20 positive control groups (heart-damaged mice were fed daily with American GNC Jiananxi coenzyme Q10, dosage according to the instructions); 20 low dose isolated and purified GABA groups (heart-damaged mice are fed with GABA content of 0.1g/L solution every day); the GABA group is separated and purified by 20 medium doses (the heart-damaged mice are fed with GABA content of 0.5/L solution every day); the GABA group was isolated and purified at 20 high doses (heart-damaged mice were fed with 1.0g/L GABA content solution per day).
The experimental method comprises the following steps: feeding pure water and American GNC Jiananxi coenzyme Q10 to heart-damaged mice, separating and purifying GABA solution with different concentrations for 14 days, and taking heart tissues to detect the morphological structure of heart cells, the change of malondialdehyde level of the heart tissues and the change of high-density lipoprotein and neurotrophic factor content in heart blood. Detecting the change of the malondialdehyde content in the heart tissue by using a malondialdehyde detection kit (Nanjing Biotechnology Co., Ltd., reference specification of the detection method); detecting the change of the content of the high-density lipoprotein by using a high-density lipoprotein detection kit (Nanjing Biotechnology Co., Ltd., reference specification of detection method); detecting the content of the cardiac nerve growth factor by using a neurotrophic factor Elisa kit (Houston corporation, USA, a reference instruction of a detection method); the cardiac cell structure is observed by conventional slice HE staining and microscopic examination.
Detection indexes are as follows: the change of the morphological structure of heart cells and the change of the contents of malondialdehyde, high-density lipoprotein and neurotrophic factor in heart tissues.
(1) The results of the morphological structure change of the heart cells are shown in fig. 1: the normal group of myocardial cells are closely arranged, the cell nucleus is positioned in the center of the cell, the cell membrane boundary is clear, the number of blood cells is large, and the blood flow is full; the density of the myocardial cells of the negative control group is reduced, the arrangement is loose, the cell membrane boundary is not clear, and the density of the blood cells is low; the heart damage phenomena of the positive control group and the group fed with 1g/L of separated and purified GABA are obviously improved, the density of cardiac muscle cells and blood cells is higher than that of the negative control group, the cell membrane boundary is clear, and the cell nucleus is positioned in the center of the cell. Therefore, the high concentration GABA (1g/L) has obvious improvement effect on the heart injury induced by NaF, leads the heart cell tissue structure to tend to be normalized, and has no obvious difference with a positive control group.
(2) The results of the change in the neurotrophic factor content in the heart homogenate showed: the content of neurotrophic factors of mice fed with 1g/L of separated and purified GABA group is increased, is close to the value of a normal group, is obviously higher than that of a negative control group, has no obvious difference with the positive control group, and the result is shown in table 1; the content of the neurotrophic factor of mice fed with 0.1g/L and 0.5g/L of GABA separated and purified groups has no significant difference with that of a negative control group; proved that GABA separated and purified from fermentation liquor of low-value fish or processing waste thereof has obvious promotion effect on the synthesis of neurotrophic factors under high concentration (1g/L), can ensure the normal play of the regulation and control function of the autonomic nervous system in the heart, and ensures the blood flow of the heart and the rhythmicity of myocardial beating.
TABLE 1 neurotrophic factor content in cardiac homogenate
1. normal group mice were fed purified water daily; 2. the negative control group is the heart-damaged model mouse which is fed with purified water every day; 3. positive control group is heart damaged model mouse fed with American GNC JIAN' ANXI coenzyme Q10 (dosage is referred to the instruction); 4. the GABA group is that GABA obtained by separating and purifying is fed to heart damaged model mice by 1g/L every day.
(3) Malondialdehyde is one of the most important products of membrane lipid peroxidation, and the generation of the malondialdehyde can also aggravate membrane damage, so that the content of the malondialdehyde is a common index in physiological researches on aging physiology and resistance of tissues and organs, and the degree of membrane lipid peroxidation can be known through the malondialdehyde so as to indirectly measure the damage degree of a membrane system and the stress resistance of the tissues and organs. The content of malondialdehyde in heart tissues of mice taking 1g/L of the separately purified GABA group is obviously lower than that of a negative control group, and no significant difference exists between the heart tissues of the mice taking the separately purified GABA group and the positive control group, and the results are shown in Table 2; there was no significant difference between the malondialdehyde content in the heart tissue of mice in the groups administered with 0.1g/L and 0.5g/L of separately purified GABA and the negative control group. Proves that the separated and purified GABA has a protective effect on heart tissues damaged by fluoride ions under a high concentration (1g/L), can reduce the damage degree of a heart cell membrane system and improve the stress resistance of the heart tissues.
TABLE 2 influence of GABA on malondialdehyde content in cardiac homogenate
Normal group mice were fed purified water daily; the negative control group is the heart-damaged model mouse which is fed with purified water every day; positive control group is heart damaged model mouse fed with American GNC JIAN' ANXI coenzyme Q10 (dosage is referred to the instruction); the GABA group is that GABA obtained by separating and purifying is fed to heart damaged model mice by 1g/L every day.
(4) The high-density lipoprotein is plasma lipoprotein for resisting atherosclerosis, is a protective factor for coronary heart disease, and is commonly called as blood vessel scavenger. The content of high-density lipoprotein in heart tissues of mice fed with 1g/L of isolated and purified GABA groups is increased, and the result is shown in Table 3; the content of high-density lipoprotein in heart tissues of mice fed with 0.1g/L and 0.5g/L of the separated and purified GABA groups has no significant difference with that of a negative control group.
TABLE 3 high Density lipoprotein content in cardiac tissue
Normal group mice were fed purified water daily; the negative control group is the heart-damaged model mouse which is fed with purified water every day; positive control group is heart damaged model mouse fed with American GNC JIAN' ANXI coenzyme Q10 (dosage is referred to the instruction); the GABA group is that GABA obtained by separating and purifying is fed to heart damaged model mice by 1g/L every day.
The test results show that GABA biologically converted from low-value fish or processing waste thereof has obvious heart protection effect.
Claims (6)
1. The application of gamma-aminobutyric acid (GABA) in preparing the medicine for protecting heart damage caused by fluorine ion induction comprises the following steps:
normalizing the tissue structure of heart cells, promoting the synthesis of neurotrophic factors, ensuring the normal performance of the regulation and control function of the autonomic nervous system in the heart, and ensuring the blood flow of the heart and the rhythmicity of the beating of cardiac muscle;
reducing the malondialdehyde content in heart tissue;
increasing the content of high density lipoprotein in heart tissue.
2. Use according to claim 1, characterized in that: the preparation method of GABA comprises the following steps: fermenting low-value fish or processing waste thereof serving as a raw material by using microorganisms with glutamate decarboxylase activity, and separating and purifying the obtained fermentation product to obtain the feed.
3. The use according to claim 2, wherein the GABA is prepared by a process comprising:
1) grinding raw material low-value fish or processing waste thereof into homogenate, adding distilled water with 5-10 times volume of homogenate into the homogenate to prepare homogenate, adding 0-5 wt% of glucose in the volume of the homogenate into the homogenate, adjusting the pH value of the solution to 4.5-6.0 to obtain a microorganism culture solution, sterilizing the culture solution, cooling, inoculating a microorganism with glutamate decarboxylase activity, and culturing at 30-40 ℃ for 24-72 hours to obtain a bacterial suspension rich in glutamate decarboxylase activity microorganism;
2) adjusting the pH value of the bacterial suspension rich in the glutamic acid decarboxylase active microorganisms obtained in the step 1) to be 5.5-6.5, adding 0.5-2 wt% of sodium glutamate in the volume of the bacterial suspension to prepare a culture solution rich in sodium glutamate, and culturing at the temperature of 35-40 ℃ for 72-120 hours to obtain fermentation liquor rich in GABA;
3) and (3) removing thallus and degreasing of GABA-rich fermentation liquor: centrifuging the fermentation liquid obtained in the step 2) for 5-20 minutes at the speed of 2000-;
4) and (3) decoloring thallus-free and grease-free fermentation liquor: decolorizing with active carbon or SD300 resin to obtain colorless transparent fermentation liquid;
5) and (3) separating and purifying GABA in colorless and transparent fermentation liquor: fully performing exchange adsorption on the 732 resin hydrogen type exchange column pretreated on the colorless transparent fermentation liquid obtained in the step 4) at the flow rate of 2-5mL/min, washing the column to pH6.0 by deionized water after adsorption, washing the column by using 100-200mL of 0.1-0.2mol/L alkaline solution, and then eluting the column by using 1-2mol/L alkaline solution at the flow rate of 2-5 mL/min; and (3) developing with ninhydrin acetone solution during the elution process, collecting effluent with ninhydrin reaction showing blue color and pH of about 6.0, detecting GABA content in each tube of effluent by high performance liquid chromatography, combining the effluent with high GABA content, and concentrating under reduced pressure to obtain separated and purified GABA.
4. Use according to claim 3, characterized in that:
the step 4) of adopting the activated carbon to decolor the thallus-free and grease-free fermentation liquor is to add 0.5 to 5 percent (w/v) of the activated carbon into the thallus-free and grease-free fermentation liquor, carry out water bath for 10 to 30 minutes at a temperature of between 60 and 90 ℃, and carry out vacuum filtration when the liquor is hot to obtain colorless and transparent fermentation liquor;
the SD300 resin decolorization is to decolorize thallus-free and grease-free fermentation liquor by adding 5-10% SD300 resin into the thallus-free and grease-free fermentation liquor, decolorizing at 20-30 ℃ for 3 hours by using a shaking table at 100rpm, and filtering to obtain colorless transparent fermentation liquor.
5. Use according to claim 3, characterized in that: and 5) concentrating under reduced pressure to 1/20-1/10 volume to obtain separated and purified GABA concentrated solution, and detecting the GABA content in the concentrated solution by using high performance liquid chromatography.
6. Use according to claim 3, characterized in that: and 5) the alkaline solution is a sodium hydroxide solution or ammonia water.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201611203181.3A CN106692123B (en) | 2016-12-23 | 2016-12-23 | Application of gamma-aminobutyric acid in preparation of heart protection pharmaceutical preparation |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201611203181.3A CN106692123B (en) | 2016-12-23 | 2016-12-23 | Application of gamma-aminobutyric acid in preparation of heart protection pharmaceutical preparation |
Publications (2)
Publication Number | Publication Date |
---|---|
CN106692123A CN106692123A (en) | 2017-05-24 |
CN106692123B true CN106692123B (en) | 2020-02-07 |
Family
ID=58903055
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201611203181.3A Active CN106692123B (en) | 2016-12-23 | 2016-12-23 | Application of gamma-aminobutyric acid in preparation of heart protection pharmaceutical preparation |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN106692123B (en) |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108841881A (en) * | 2018-06-19 | 2018-11-20 | 中国科学院海洋研究所 | A method of γ-aminobutyric acid is prepared by raw material of sea robin |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103966274A (en) * | 2013-02-05 | 2014-08-06 | 中国科学院海洋研究所 | Method for biotransformation production of gamma-aminobutyric acid with aquatic products and processing leftovers thereof as raw materials |
CN104825434A (en) * | 2015-03-31 | 2015-08-12 | 中国科学院海洋研究所 | Preparation with hypothyroidism resisting effect, and preparation method thereof |
CN104840800A (en) * | 2015-06-08 | 2015-08-19 | 福州乾正药业有限公司 | Composition of bamboo leaf flavonoid and gamma-aminobutyric acid as well as preparation method and applications thereof |
CN104922145A (en) * | 2015-05-25 | 2015-09-23 | 福州乾正药业有限公司 | Composition of gamma-aminobutyric acid and chitosan oligosaccharide, as well as preparation method and applications of composition |
-
2016
- 2016-12-23 CN CN201611203181.3A patent/CN106692123B/en active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103966274A (en) * | 2013-02-05 | 2014-08-06 | 中国科学院海洋研究所 | Method for biotransformation production of gamma-aminobutyric acid with aquatic products and processing leftovers thereof as raw materials |
CN104825434A (en) * | 2015-03-31 | 2015-08-12 | 中国科学院海洋研究所 | Preparation with hypothyroidism resisting effect, and preparation method thereof |
CN104922145A (en) * | 2015-05-25 | 2015-09-23 | 福州乾正药业有限公司 | Composition of gamma-aminobutyric acid and chitosan oligosaccharide, as well as preparation method and applications of composition |
CN104840800A (en) * | 2015-06-08 | 2015-08-19 | 福州乾正药业有限公司 | Composition of bamboo leaf flavonoid and gamma-aminobutyric acid as well as preparation method and applications thereof |
Non-Patent Citations (5)
Title |
---|
Prevention of stress-induced disturbances of contractility of nonischemic regions of the heart in myocardial infarction by gamma-hydroxybutyric acid;R.S. Dosmagambetova;《Bulletin of experimental biology and medicine》;19831231;第96卷(第6期);第1679-1681页 * |
γ-氨基丁酸的抗心律失常作用;常洁琴等;《中国药学杂志》;19941231;第752页 * |
大剂量γ-氨基丁酸和二巯丙基磺酸钠联合维生素B6对毒鼠强剂型中毒大鼠肝、脑、心的影响;谢海;《中国优秀硕士学位论文全文数据库 医药卫生科技辑》;20110215;第26页第2段、第27页第2段、第29页第1段 * |
常洁琴等.γ-氨基丁酸的抗心律失常作用.《中国药学杂志》.1994,第752页. * |
氟中毒对心脏损伤机理的研究;杨霞等;《中华地方病学杂志》;20140920;第33卷(第5期);第20-22页 * |
Also Published As
Publication number | Publication date |
---|---|
CN106692123A (en) | 2017-05-24 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2020177390A1 (en) | Method for preparing l-ergothioneine-containing cosmetic stock solution by means of fermenting hericium erinaceus | |
AU2004276123B2 (en) | Fermentation and culture method, fermented plant extract, fermented plant extract powder and composition containing the fermented plant extract | |
CN103966274A (en) | Method for biotransformation production of gamma-aminobutyric acid with aquatic products and processing leftovers thereof as raw materials | |
Nakano et al. | Neutralization of microcystin shock in mice by tumor necrosis factor alpha antiserum | |
CN110800871A (en) | Application of schizochytrium limacinum powder in improving DHA content in ruminant milk | |
CN116555129B (en) | Lactobacillus gasseri BDUP, application and product thereof | |
CN115322932B (en) | Lactobacillus plantarum with anti-alcohol and sobering-up capabilities and application thereof | |
US20130217085A1 (en) | Methods for improving fermentation yield of polyunsaturated fatty acids | |
CN107252475B (en) | The application of natural host defense peptide Alligatorin4 | |
CN101057873A (en) | Method for preparing hepatitis B virus resisting maggot protein powder and its maggot oil and fatty acid | |
CN111560321A (en) | Efficient fermentation process of rhodotorula benthica | |
CN110656151A (en) | Method for improving kidney-tonifying and yang-strengthening effects of sea cucumber | |
CN109806383A (en) | A kind of sea eel peptide promotes the application in immune food, drug or health care product in preparation | |
CN106692123B (en) | Application of gamma-aminobutyric acid in preparation of heart protection pharmaceutical preparation | |
CN102356879A (en) | Functional peptide-reinforced healthcare food | |
CN107854508B (en) | Composition for relieving alcoholic liver injury and preparation method thereof | |
CN100490829C (en) | Selenium-rich hedgehog fungus extract preparation for treating colitis, gastritis and stomach cancer and its preparing process | |
CN104825434A (en) | Preparation with hypothyroidism resisting effect, and preparation method thereof | |
CN106667987A (en) | Application of gamma-aminobutyric acid (GABA) in preparing liver-protecting medicinal preparation | |
CN104404103B (en) | Zymotic fluid prepared by a kind of method and this method for improving Polysaccharide from Portulaca oleracea | |
TWI689593B (en) | Method for preparing high-yield γ-aminobutyric acid | |
CN103757076A (en) | Method for increasing valence of glutathione in saccharomyces cerevisiae fermentation liquor by utilizing oxygen carrier | |
CN117511825B (en) | Application of segment fermentation inoculant of Wittman coagulans IOB502 in alcoholic liver injury | |
KR102681818B1 (en) | Method for preparing spirulina extract with increased branched chain amino acid content | |
KR20240100578A (en) | Novel Pediococcus genus strain having ability to reduce off-flavor in aquatic products and composition for reducing off-flavor in aquatic products containing the same |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |