CN108159037A - A kind of unsaturated fatty-acid compositions and its application for being used to improve neuroprotective function - Google Patents
A kind of unsaturated fatty-acid compositions and its application for being used to improve neuroprotective function Download PDFInfo
- Publication number
- CN108159037A CN108159037A CN201711468538.5A CN201711468538A CN108159037A CN 108159037 A CN108159037 A CN 108159037A CN 201711468538 A CN201711468538 A CN 201711468538A CN 108159037 A CN108159037 A CN 108159037A
- Authority
- CN
- China
- Prior art keywords
- dha
- epa
- unsaturated fatty
- cell
- ratio
- 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.)
- Pending
Links
- 239000000203 mixture Substances 0.000 title claims abstract description 30
- 235000021122 unsaturated fatty acids Nutrition 0.000 title claims abstract description 23
- 150000004670 unsaturated fatty acids Chemical class 0.000 title claims abstract description 23
- 230000000324 neuroprotective effect Effects 0.000 title claims abstract description 21
- 235000013402 health food Nutrition 0.000 claims abstract description 10
- 235000013305 food Nutrition 0.000 claims description 11
- 239000003814 drug Substances 0.000 claims description 10
- 230000000694 effects Effects 0.000 abstract description 17
- 238000002156 mixing Methods 0.000 abstract 1
- MBMBGCFOFBJSGT-KUBAVDMBSA-N all-cis-docosa-4,7,10,13,16,19-hexaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCC(O)=O MBMBGCFOFBJSGT-KUBAVDMBSA-N 0.000 description 187
- 235000020669 docosahexaenoic acid Nutrition 0.000 description 94
- 229940090949 docosahexaenoic acid Drugs 0.000 description 93
- JAZBEHYOTPTENJ-JLNKQSITSA-N all-cis-5,8,11,14,17-icosapentaenoic acid Chemical compound CC\C=C/C\C=C/C\C=C/C\C=C/C\C=C/CCCC(O)=O JAZBEHYOTPTENJ-JLNKQSITSA-N 0.000 description 81
- 235000020673 eicosapentaenoic acid Nutrition 0.000 description 81
- 229960005135 eicosapentaenoic acid Drugs 0.000 description 81
- JAZBEHYOTPTENJ-UHFFFAOYSA-N eicosapentaenoic acid Natural products CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O JAZBEHYOTPTENJ-UHFFFAOYSA-N 0.000 description 81
- 210000004027 cell Anatomy 0.000 description 50
- 230000014509 gene expression Effects 0.000 description 22
- 230000006870 function Effects 0.000 description 18
- 230000003833 cell viability Effects 0.000 description 16
- 238000012545 processing Methods 0.000 description 15
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 12
- 108010025020 Nerve Growth Factor Proteins 0.000 description 11
- 230000006698 induction Effects 0.000 description 11
- 108020004999 messenger RNA Proteins 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 11
- MWUXSHHQAYIFBG-UHFFFAOYSA-N Nitric oxide Chemical compound O=[N] MWUXSHHQAYIFBG-UHFFFAOYSA-N 0.000 description 10
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 10
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 10
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 9
- 239000002253 acid Substances 0.000 description 9
- 102000004169 proteins and genes Human genes 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 239000003963 antioxidant agent Substances 0.000 description 8
- 230000003078 antioxidant effect Effects 0.000 description 8
- 235000006708 antioxidants Nutrition 0.000 description 8
- 238000002474 experimental method Methods 0.000 description 8
- 108700000707 bcl-2-Associated X Proteins 0.000 description 7
- 102000055102 bcl-2-Associated X Human genes 0.000 description 7
- 229940079593 drug Drugs 0.000 description 7
- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 6
- 108090000715 Brain-derived neurotrophic factor Proteins 0.000 description 6
- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 6
- 102000015336 Nerve Growth Factor Human genes 0.000 description 6
- 230000003110 anti-inflammatory effect Effects 0.000 description 6
- 230000004069 differentiation Effects 0.000 description 6
- 229960003180 glutathione Drugs 0.000 description 6
- 102000004219 Brain-derived neurotrophic factor Human genes 0.000 description 5
- 108090000397 Caspase 3 Proteins 0.000 description 5
- 102000003952 Caspase 3 Human genes 0.000 description 5
- 102000007072 Nerve Growth Factors Human genes 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 239000003900 neurotrophic factor Substances 0.000 description 5
- 230000036542 oxidative stress Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 206010029350 Neurotoxicity Diseases 0.000 description 4
- 101150056950 Ntrk2 gene Proteins 0.000 description 4
- 206010044221 Toxic encephalopathy Diseases 0.000 description 4
- 239000003795 chemical substances by application Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 230000004112 neuroprotection Effects 0.000 description 4
- 230000007135 neurotoxicity Effects 0.000 description 4
- 231100000228 neurotoxicity Toxicity 0.000 description 4
- 235000020660 omega-3 fatty acid Nutrition 0.000 description 4
- 230000004224 protection Effects 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 108010085238 Actins Proteins 0.000 description 3
- 102000007469 Actins Human genes 0.000 description 3
- 108020004635 Complementary DNA Proteins 0.000 description 3
- 102000004127 Cytokines Human genes 0.000 description 3
- 108090000695 Cytokines Proteins 0.000 description 3
- 101150111783 NTRK1 gene Proteins 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 3
- 230000006907 apoptotic process Effects 0.000 description 3
- 210000004556 brain Anatomy 0.000 description 3
- 238000010804 cDNA synthesis Methods 0.000 description 3
- 230000005779 cell damage Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 235000021323 fish oil Nutrition 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 235000013336 milk Nutrition 0.000 description 3
- 239000008267 milk Substances 0.000 description 3
- 210000004080 milk Anatomy 0.000 description 3
- 210000002569 neuron Anatomy 0.000 description 3
- 239000008194 pharmaceutical composition Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 229930002330 retinoic acid Natural products 0.000 description 3
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 2
- 101150035467 BDNF gene Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 230000004913 activation Effects 0.000 description 2
- 125000001931 aliphatic group Chemical group 0.000 description 2
- 238000003149 assay kit Methods 0.000 description 2
- 239000002775 capsule Substances 0.000 description 2
- 230000030833 cell death Effects 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 235000013365 dairy product Nutrition 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 235000005911 diet Nutrition 0.000 description 2
- 230000037213 diet Effects 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 230000004064 dysfunction Effects 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 239000000499 gel Substances 0.000 description 2
- 235000003642 hunger Nutrition 0.000 description 2
- 230000002757 inflammatory effect Effects 0.000 description 2
- 210000000274 microglia Anatomy 0.000 description 2
- 230000001537 neural effect Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 108090000765 processed proteins & peptides Proteins 0.000 description 2
- 230000000770 proinflammatory effect Effects 0.000 description 2
- 230000002633 protecting effect Effects 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 108020003175 receptors Proteins 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 230000009897 systematic effect Effects 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 229960001727 tretinoin Drugs 0.000 description 2
- DVSZKTAMJJTWFG-SKCDLICFSA-N (2e,4e,6e,8e,10e,12e)-docosa-2,4,6,8,10,12-hexaenoic acid Chemical compound CCCCCCCCC\C=C\C=C\C=C\C=C\C=C\C=C\C(O)=O DVSZKTAMJJTWFG-SKCDLICFSA-N 0.000 description 1
- MIDXCONKKJTLDX-UHFFFAOYSA-N 3,5-dimethylcyclopentane-1,2-dione Chemical compound CC1CC(C)C(=O)C1=O MIDXCONKKJTLDX-UHFFFAOYSA-N 0.000 description 1
- UWSONZCNXUSTKW-UHFFFAOYSA-N 4,5-Dimethylthiazole Chemical compound CC=1N=CSC=1C UWSONZCNXUSTKW-UHFFFAOYSA-N 0.000 description 1
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- GZJLLYHBALOKEX-UHFFFAOYSA-N 6-Ketone, O18-Me-Ussuriedine Natural products CC=CCC=CCC=CCC=CCC=CCC=CCCCC(O)=O GZJLLYHBALOKEX-UHFFFAOYSA-N 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 208000024827 Alzheimer disease Diseases 0.000 description 1
- 208000037259 Amyloid Plaque Diseases 0.000 description 1
- 102000002659 Amyloid Precursor Protein Secretases Human genes 0.000 description 1
- 108010043324 Amyloid Precursor Protein Secretases Proteins 0.000 description 1
- 241000972773 Aulopiformes Species 0.000 description 1
- 241001474374 Blennius Species 0.000 description 1
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 102000008946 Fibrinogen Human genes 0.000 description 1
- 108010049003 Fibrinogen Proteins 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 235000010469 Glycine max Nutrition 0.000 description 1
- 244000068988 Glycine max Species 0.000 description 1
- 108020005004 Guide RNA Proteins 0.000 description 1
- 101710088172 HTH-type transcriptional regulator RipA Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 102000004889 Interleukin-6 Human genes 0.000 description 1
- 108090001005 Interleukin-6 Proteins 0.000 description 1
- 241000699660 Mus musculus Species 0.000 description 1
- 101150064037 NGF gene Proteins 0.000 description 1
- 206010029260 Neuroblastoma Diseases 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 102000003992 Peroxidases Human genes 0.000 description 1
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 1
- 244000046052 Phaseolus vulgaris Species 0.000 description 1
- 229920009405 Polyvinylidenefluoride (PVDF) Film Polymers 0.000 description 1
- 108010009736 Protein Hydrolysates Proteins 0.000 description 1
- 244000018633 Prunus armeniaca Species 0.000 description 1
- 235000009827 Prunus armeniaca Nutrition 0.000 description 1
- 239000012083 RIPA buffer Substances 0.000 description 1
- 238000002123 RNA extraction Methods 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 206010039966 Senile dementia Diseases 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 230000000996 additive effect Effects 0.000 description 1
- 230000002776 aggregation Effects 0.000 description 1
- 238000004220 aggregation Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 230000002424 anti-apoptotic effect Effects 0.000 description 1
- 239000002260 anti-inflammatory agent Substances 0.000 description 1
- 229940121363 anti-inflammatory agent Drugs 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 230000001741 anti-phlogistic effect Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012752 auxiliary agent Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000009835 boiling Methods 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 235000013736 caramel Nutrition 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 230000007248 cellular mechanism Effects 0.000 description 1
- 210000003710 cerebral cortex Anatomy 0.000 description 1
- 235000013351 cheese Nutrition 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 210000002932 cholinergic neuron Anatomy 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 238000003776 cleavage reaction Methods 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 230000019771 cognition Effects 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000008878 coupling Effects 0.000 description 1
- 238000010168 coupling process Methods 0.000 description 1
- 238000005859 coupling reaction Methods 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 239000002781 deodorant agent Substances 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- KAUVQQXNCKESLC-UHFFFAOYSA-N docosahexaenoic acid (DHA) Natural products COC(=O)C(C)NOCC1=CC=CC=C1 KAUVQQXNCKESLC-UHFFFAOYSA-N 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 238000006911 enzymatic reaction Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000012894 fetal calf serum Substances 0.000 description 1
- 229940012952 fibrinogen Drugs 0.000 description 1
- 235000019688 fish Nutrition 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 235000013355 food flavoring agent Nutrition 0.000 description 1
- 235000003599 food sweetener Nutrition 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 210000001320 hippocampus Anatomy 0.000 description 1
- 235000012907 honey Nutrition 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000037189 immune system physiology Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000008798 inflammatory stress Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 229940100601 interleukin-6 Drugs 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 235000021388 linseed oil Nutrition 0.000 description 1
- 239000000944 linseed oil Substances 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 235000020667 long-chain omega-3 fatty acid Nutrition 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000002844 melting Methods 0.000 description 1
- 230000008018 melting Effects 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000004770 neurodegeneration Effects 0.000 description 1
- 230000019581 neuron apoptotic process Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 235000014571 nuts Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 108040007629 peroxidase activity proteins Proteins 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 150000003904 phospholipids Chemical class 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000013139 quantization Methods 0.000 description 1
- 238000010814 radioimmunoprecipitation assay Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- CZCBTSFUTPZVKJ-UHFFFAOYSA-N rose oxide Chemical compound CC1CCOC(C=C(C)C)C1 CZCBTSFUTPZVKJ-UHFFFAOYSA-N 0.000 description 1
- 239000012146 running buffer Substances 0.000 description 1
- 235000019515 salmon Nutrition 0.000 description 1
- 230000007017 scission Effects 0.000 description 1
- 230000028327 secretion Effects 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 239000002002 slurry Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000003765 sweetening agent Substances 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- -1 tetrazolium salt Chemical class 0.000 description 1
- 210000001519 tissue Anatomy 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 238000011830 transgenic mouse model Methods 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
- 235000021081 unsaturated fats Nutrition 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 108700026220 vif Genes Proteins 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
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
- A61K31/20—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids
- A61K31/202—Carboxylic acids, e.g. valproic acid having a carboxyl group bound to a chain of seven or more carbon atoms, e.g. stearic, palmitic, arachidic acids having three or more double bonds, e.g. linolenic
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
- A23L33/12—Fatty acids or derivatives thereof
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Public Health (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Veterinary Medicine (AREA)
- Epidemiology (AREA)
- Mycology (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
本发明涉及一种用于提高神经保护功能的不饱和脂肪酸组合物，其特征在于：EPA:DHA的摩尔比为1:2。本发明还涉及一种用于提高神经保护功能的保健食品，其特征在于：含有权利要求1所述的不饱和脂肪酸组合物，所述不饱和脂肪酸组合物中，EPA:DHA的摩尔比为1:2。本发明通过调整比例EPA和DHA组合，达到最优的神经保护效果，从中确定了最为有效的比例组合。并可通过人工调和达到最优比例配方，从而提供产品功能效果。
The invention relates to an unsaturated fatty acid composition for improving neuroprotective function, which is characterized in that the molar ratio of EPA:DHA is 1:2. The present invention also relates to a health food for improving neuroprotective function, characterized in that: it contains the unsaturated fatty acid composition according to claim 1, and in the unsaturated fatty acid composition, the molar ratio of EPA:DHA is 1 :2. The present invention achieves the optimal neuroprotective effect by adjusting the ratio of EPA and DHA, and determines the most effective ratio combination. And it can achieve the optimal ratio formula through manual blending, so as to provide product functional effect.
Description
Technical field
The present invention relates to a kind of for improving unsaturated fatty-acid compositions and its application of neuroprotective function, and then,
It is related to health food, food nutrition, drug field.
Background technology
Epidemiological survey has proven to the low incidence of AD (senile dementia) and the diet ω -3PUFAs of intake high-content,
Mainly eicosapentaenoic acid (EPA) is related to docosahexaenoic acid (DHA).ω -3 long-chain polyunsaturated fatty acids (ω -
It is essential nutriment 3PUFAs) as the component part of neuron membrane, main make is risen to human organ and its function
With.They participate in inflammation and immunological process and hormone control.In addition, they participate in development and the function of brain.
Numerous studies have proven to oxidative stress and can result in damage and the dysfunction of neuron with long-term chronic inflammation, are
Depression or the inducement of AD morbidities.
Several evidences show that the presence of inflammatory cytokine in AD increases, such as TNF-α level increases in AD patients serums
The presence of (Khemka etc., 2014) and interleukin-6 mRNA in APPsw transgenic mice Tg2576 hippocampus and cortex with
And around the increase of the microglia cell of the activation of senile plaque in cerebral cortex.
The microglia cell of activation can not only generate proinflammatory cytokine, but also can generate free radicals an oxidation
Nitrogen and superoxide anion.These groups induce the neurodegeneration event similar to AD together with TNF-α secretion object.Research shows that
A β in SH-SY5Y cells25-35Toxicity and ROS and NO releases and the enhancing of oxidative damage it is related, it is quick to have raised redox
The transcription factor of sense such as NF- κ B, this is oxidation and an important factor for inflammatory reaction in AD.
In previous studies, it was recently reported that neuroinflamation can reduce the level of neurotrophic factor such as NGF and BDNF.Thing
In reality, it is reported in the concentration of neurotrophic factor and function of receptors increased and decreased in AD patient.Neurotrophy system participates in
With neure growth, survival and the relevant many physiology courses of plasticity.It also reported cholinergic neuron nerve to occur in AD
Dysfunction.
ω -3PUFAs are related to the structure and function that cell membrane phospholipid is formed in brain, and are considered the lifting in cognitive process
It acts on.Secondly, it has been found that ω -3PUFAs have anti-oxidant and anti-inflammatory effect.Particularly in the brain of aging, this spy
Property potentially contributes to the protection of neuron and prevents cell death.
At present, the fish oil comprising unrighted acid or linseed oil have been widely used in health food.However facing
In bed and experiment, the effect of different unrighted acids is not consistent：There is good anti-inflammatory, the anti-oxidant, neuroprotection of display,
There is invalid report.These different effects may it is related with its separate sources insatiable hunger aliphatic acid used or with its source difference institute
The ratio difference containing DHA with EPA leads to the difference of result.
Invention content
The purpose of the present invention is to provide a kind of for improving the unsaturated fatty-acid compositions of neuroprotective function, comprising
EPA and DHA, it is characterised in that：EPA:The molar ratio of DHA is 1:2.
It is an object of the invention to also provide a kind of health food for being used to improve neuroprotective function, it is characterised in that：
Containing unsaturated fatty-acid compositions of the present invention, in the unsaturated fatty-acid compositions, EPA:The molar ratio of DHA is
1:2。
The present invention further provides the unsaturated fatty-acid compositions in the drug for improving neuroprotective function
Using.
The present invention provides application of the unsaturated fatty-acid compositions in the food for improving neuroprotective function.
The present inventor has studied the anti-oxidant, anti-inflammatory and neural of different proportion DHA and EPA ratio combination by systematic comparison
Protecting effect therefrom determines maximally efficient ratio combination.And ratio formula can be optimal by manually reconciling, so as to carry
For product function effect.
Description of the drawings
Fig. 1 show to prepare for improve neuroprotective function unsaturated fatty-acid compositions flow chart.
The influence that cell viability EPA, DHA of Fig. 2 a various concentrations beta induced to A reduces.
The influence that Fig. 2 b EPA, DHA and its different proportion cell viability beta induced to A reduce.
The protective effect of Fig. 2 c EPA, DHA and its different proportion cellular damage beta induced to A.
The influence of Fig. 3 a EPA, DHA and its different proportion the ROS level variation beta induced to A.
The influence of Fig. 3 b EPA, DHA and its different proportion the NO level variation beta induced to A.
The influence of Fig. 3 c EPA, DHA and its different proportion the GSH level variation beta induced to A.
Fig. 4 a EPA, DHA and its different proportion the TNF-α mRNA beta induced on A express increased influence.
The horizontal increased influence of Fig. 4 b EPA, DHA and its different proportion TNF-α beta induced on A.
The influence of Fig. 5 a EPA, DHA and its different proportion the NGF mRNA expression beta induced to A.
The influence of Fig. 5 b EPA, DHA and its different proportion the BDNF mRNA expression beta induced to A.
The influence of Fig. 5 c EPA, DHA and its different proportion the NGF level beta induced to A.
The influence of Fig. 5 d EPA, DHA and its different proportion the BDNF level beta induced to A.
Fig. 6 a EPA, DHA and its different proportion Bax beta induced to A：The influence of Bcl-2 ratios.
The influence of Fig. 6 b EPA, DHA and its different proportion the Caspase-3 protein expression beta induced to A.
Specific embodiment
Below in conjunction with the specific embodiment of the invention, technical scheme of the present invention is verified, the embodiment verified
Only section Example of the invention.Based on the embodiments of the present invention, this field researcher is not making any creation
Property labour under the premise of the other embodiment that is obtained, shall fall within the protection scope of the present invention.
In the unsaturated fatty-acid compositions for being used to improve neuroprotective function of the present invention, EPA:The molar ratio of DHA is
1:2。
EPA, DHA wherein in composition are commercially available sterling, and the preparation flow of composition is as shown in Figure 1：It surveys respectively first
The influence of the cell viability of the AD cell models of EPA and DHA the SH-SY5Y cell beta induced to A of various concentration is tried, so as to really
Surely the dose concentration to shield, EPA and DHA processing cells then again in varing proportions, detects the thin of SH-SY5Y cells
Born of the same parents' vigor, Antioxidant Indexes：ROS, NO, GHA, anti-inflammatory index：TNF-α, anti-apoptotic index Bc1, Bax, Caspase-3, so as to
Determine EPA：The molar ratio of DHA is 1:Neuroprotective effect is optimal when 2, only need to manually allocate EPA and DHA to required ratio,
It is uniformly mixed using conventional method and obtains the composition.
It is appreciated that the unsaturated fatty-acid compositions of the present invention, can individually eat as health food, can also add
Enter into other food mixed edible or applied in the drug for improving neuroprotective function is prepared.
The health food for being used to improve neuroprotective function of the present invention, contains the insatiable hunger described in the first aspect of the present invention
And aliphatic acid composition, in the unsaturated fatty-acid compositions, EPA:The molar ratio of DHA is 1:2.
The health food can be the form of ordinary food, can also use tablet, capsule, pill, lyophilized form or
Any suitable administration form of person.
Health food in the scope of the invention may include the additive below one or more：Preservative, solubilizer, stabilization
Agent, wetting agent, emulsifier, sweetener, colorant, flavoring agent, deodorant, buffer solution, coating agent, antioxidant, suspending agent,
Auxiliary agent, excipient and diluent.
The present invention further provides the unsaturated fatty-acid compositions in the drug for improving neuroprotective function
Using.
The unsaturated fatty-acid compositions containing EPA and DHA can be used in pharmaceutical composition so that rich in drug
Containing EPA and DHA.
The convenient form of the pharmaceutical composition can be tablet for oral administration, pill, capsule, syrup
Agent, pulvis or granule；Sterile parenteral or subcutaneous solution for parenterai administration, suspension or the bolt for rectally
Agent, all these is all well known in the art.
Specific dosage level and dose frequency can be variation for any specific patient, will depend on more
Kind of factor, the metabolic stability of activity, this compound activity including used specific compound and duration, the age,
Weight, general health, gender, diet, the mode of administration and time, the rate of excretion, the combining of drug, specific disease
The severity of disease and the treatment of single progress.The medicament and/or pharmaceutical composition of the present invention can be according to daily 1~10 time
Administration, such as once or twice daily.For oral and parenteral administration patient, the dosage level of medicament can be single dose
Amount or separated dosage.
The present invention provides application of the unsaturated fatty-acid compositions in the food for improving neuroprotective function.
The composition of the present invention can be used in food industry so that food (such as grain food, dairy products, soya-bean oil, beans
Slurry) in be rich in EPA and DHA.
The cereal foods can have different shape or appearance forrns.For example, stick, cake can be made into
Or macaroon or its can also be oatmeal shape, sheet or rodlike.It can individually eat or with dairy products such as milk, acid
Milk or cottage cheese etc. are eaten together.
It should be understood that essence can also be added in food, such as honey, fruit, chocolate, caramel, nut, apricot
Benevolence, yoghurt flavours or combination.
Current market sales of unsaturated fat acid product also has largely from deep sea fish oil from plant, various
The extraction of algae, ingredient is different, and especially there are notable differences in DHA and EPA ratios.It is such as common from gold in the market
Fish oil DHA in prosperous fish or dog salmon：EPA is close to 1:2；And the unrighted acid in seaweed then contains higher DHA ratios
Example, this leads to health effect, and there are bigger differences.And so far, it there is no to DHA and EPA ratios group in unrighted acid
It closes in anti-oxidant, anti-inflammatory, the research in terms of neuroprotection, it is unclear between the variation of the two ratio and effect, therefore also without most
Ratio of greater inequality example formula and product.
The present inventor has studied the anti-oxidant, anti-inflammatory and neural of different proportion DHA and EPA ratio combination by systematic comparison
Protecting effect therefrom determines maximally efficient ratio combination.And ratio formula can be optimal by manually reconciling, so as to really
Protect product function effect.Be it in health food or drug with providing reference.
A β be by amyloid precusor protein (APP) through β-and the protein hydrolysate of gamma-secretase under pathological conditions.A
β25-35It is a kind of synthetic peptide for the overall length A β for corresponding to 25-35 amino acid, there is identical beta sheet structure, and keep overall length
Aβ1-42Complete toxicity.The peptide shows to form the rapid aggregation property of stable fibrinogen, and has god immediately in dissolving
Through toxicity.The present invention uses A β25-35The SH-SY5Y cells of differentiation are damaged as AD models, more different EPA/DHA ratios pair
A β in SH-SY5Y cells25-35The potential neuroprotection of the neurotoxicity of induction.
At present, without any experiment respectively to EPA, DHA or a certain proportion of combinations are studied and in same experiment
More different ω -3PUFAs.The present invention studies the combination of individual EPA and DHA and its different proportion to A β25-35The AD of induction
The effect of cell model.Cell viability is measured, to compare EPA, the combination of DHA or its various ratio is to A β25-35The AD of induction
The influence of the neurotoxicity of cell model and their potential synergistic effect.In addition, measure oxidative stress, proinflammatory cytokines because
The level of son and neurotrophic factor, to analyze EPA, DHA or combinations may be beneficial to the cell and molecular mechanism of AD.And
The EPA/DHA of different proportion adjusts the ability of the expression of apoptosis-associated genes.
In the present invention, " FAs " refers to the EPA of various concentration, DHA or the combination of its different proportion.
Embodiment
Human neuroblastoma cell system SH-SY5Y cells all-trans retinoic acid (RA) is handled 7-8 days and is divided completely
Turn to human neure like cell.In the last day of differentiation, cell starts to test.In A β25-35The SH- of the differentiation of induction
EPA and DHA is tested in the AD cell models of SY5Y cells in 6,12,25,50,100 μM of influences to cell viability.Wherein occur
Slight but significant attenuation A β25-35The optimal dose of point selected as EPA and DHA that the cell viability of induction reduces and culture continue
Time.Then carry out following seven groups of researchs：(i) (culture medium) is compareed, (ii) A β25-35(add A β25-35Culture medium), (iii) A β
+ EPA (is pre-processed, then with A β with EPA25-35Processing), (iv) A β+DHA (are pre-processed, then with A β with DHA25-35Processing) and
(v-vii) A β+EPA+DHA are (respectively with 2:1,1:1 and 1:2 EPA+DHA (totally 25 μM) processing, then with A β25-35Processing).
Add A β25-35Before, cell EPA, DHA, combination thereof or control solvent are pre-processed 12 hours.Add in A β25-35(eventually
A concentration of 20 μM) after, cell is incubated 24 hours again.Then, the cell viability in SH-SY5Y cells, oxidative stress, inflammatory are studied
Cytokine TNF-α, neurotrophic factor and Apoptosis.
In the present embodiment, it as reagent, uses>Eicosapentaenoic acid (the EPA of 99% purity；20：5, n-3) and 20
Two carbon acid (DHA；22：6, n-3) sodium salt comes from Sigma-Aldrich companies.By FAs dissolvings in the medium, in nitrogen
It is divided into aliquot under air-flow, and is preserved at -80 DEG C until using.
In the present embodiment, SH-SY5Y comes from ATCC (CRL-2266, Lot.61983120).Cell is being contained 10%
Fetal calf serum (FBS,Canada) and 1% Pen .- Strep DMEM/F12 culture mediums (Add and take
Ventilation 75-cm greatly)2It is cultivated in culture bottle.It is dense with end in the DMEM/F12 containing 3%FBS (culture medium is replaced for every 2 days)
RA (Sigma Aldrich, Canada) the processing SH-SY5Y cells spent for 10 μM are divided into complete human neure for 7-8 days
Like cell.
In the present embodiment, using the 3- (4,5- dimethylthiazole -2) -2 for measuring cell proliferation rate and cell viability,
5- diphenyltetrazolium bromides (MTT) measure cell viability.By cell inoculation in 96 orifice plates, 90 are added in into each hole
μ L cell suspending liquids.After experiment process, cell viability is detected with MTT (ATCC) according to the manufacturer's instructions.Use enzyme mark
Instrument (BioTek, USA) measures optical density in 570nm.The absorbance of control group is considered as the 100% of cell viability.
As detected by mtt assay, A β25-35It lives in the cell for the SH-SY5Y cells that 20 μM significantly reduce differentiation in 24 hours
Power (p<0.01, Fig. 2 a).However, significantly reduce A β with (6-100 μM) pretreatment of EPA or DHA of various concentration25-35With agent
The reduction of cell viability caused by measuring dependence mode, and 6 in EPA, 12 (p>0.05), 25 (p<0.05), 50 μM of (p<
And 100 μM of (p 0.01)<0.05) 6 (p or in DHA<0.05), 12-50 (p<And 100 μM of (p 0.01)<0.05) (Fig. 2 a).
Compared with EPA, the effect of DHA seems more stronger than effects of the EPA under same dose.On the basis of these results, subsequent
In the measure of SH-SY5Y cells progress oxidative stress, inflammation and apoptosis, we select 25 μM as EPA in different proportion processing
With the accumulated dose of DHA combinations.
In the present embodiment, it since MTT is measured to cell quantity sensitivity, is influenced by cell Proliferation and cell viability,
It you must use another detection method and confirm result.Come using CytoTox-96 assay kits (Promega, Canada)
Total release of cytoplasm lactic dehydrogenase (LDH) in culture medium is assessed, this is the result of cell integrity damage.The measure is based on
From 2-P- (iodophenyl) -3- (p-nitrophenyl) -5- phenyltetrazoles chloride (INT, tetrazolium salt) to the coupling of formazan product
Enzymatic conversion, enzyme reaction is discharged from cell by LDH and the diaphorase in substrate mixture is measured is catalyzed.Pass through enzyme mark
Instrument reads absorbance at 490nm.Every group of mean light absorbency is normalized to the percentage of control value.
In order to test different EPA/DHA ratios in neuroprotection have influence in various degree it is assumed that test below
It is middle to be combined using following different FAs：EPA/DHA 2：1, EPA/DHA 1：1 or EPA/DHA 1：2.Fig. 2 b's the results show that
With A β25-35Group is compared, in all proportions test, different EPA/DHA ratios significantly (p<0.05) cell viability is improved.Protection
SH-SY5Y cells are from A β25-35The FAs effect of the neurotoxicity of induction is：EPA<2：1EPA/DHA<DHA≤1：1EPA/DHA<
1：2EPA/DHA.
Measure (it is the index of cell death) is discharged by LDH and further demonstrates different proportion EPA/DHA to A β25-35
The protective effect (Fig. 2 c) of the SH-SY5Y cellular damages of induction.With reference to said determination as a result, we can obtain knot for certain
By, EPA, DHA and combinations thereof can to varying degrees effective protection differentiation SH-SY5Y cells from A β25-35Induction
Cellular damage, the EPA of best raising cell viability：The ratio of DHA is 1：2EPA/DHA.
In the present embodiment, SH-SY5Y cells are inoculated in 96 orifice plates, 200 μ L cell suspending liquids are added in into each hole.
After experiment process, according to the manufacturer's instructions, with ROS kits in fluorecyte (Sigma Aldrich) quantization cell
The level of interior ROS.Using fluorescence microplate reader (Reader Synergy HT, BioTek Instruments, the U.S.), with lex=
650/lem=675nm fluorescence intensities.According to the manufacturer's instructions, by Griess reagent systems, (Promega adds and takes
Cell intracellular nitric oxide (NO) yield is measured greatly).Absorbance is measured at 540nm using microplate reader.
By SH-SY5Y cell inoculations in 6 orifice plates, and 2mL cell suspending liquids are added in into each hole.In experiment process
Afterwards, according to the manufacturer's instructions, GSH concentration is measured with glutathione assay kit (Sigma Aldrich).Fluorescence intensity
It is measured with fluorimeter reader, excitation wavelength 390nm, launch wavelength 478nm.
Fig. 3 a illustrate A β25-35ROS fluorescence (p is dramatically increased than control group<0.01).When with A β25-35When group compares, individually
(p is significantly reduced using the ROS fluorescence of the combination of EPA and DHA under EPA and all test ratios<0.05).However, in DHA groups
ROS fluorescence does not find significant difference.Protect SH-SY5Y cells from A β25-35The FAs effect for inducing increased ROS fluorescence is：
DHA<1：2EPA/DHA<EPA<1：1EPA/DHA≤2:1EPA/DHA.
As shown in Figure 3b, when individually with A β25-35When handling cell, observe that nitrate levels dramatically increase about 40.29%
(p<0.05).However, with A β25-35Group is compared, and the nitrate levels of EPA processing are used alone slightly but do not significantly reduce (p<
0.05).With A β25-35Group compares, and DHA and the processing of all EPA/DHA processing groups, the equal horizontal significance difference of nitrate-free is used alone
It is different.
It is being shown in Fig. 3 c the result shows that, A β25-35The GSH contents of damaging cells significantly (p<0.05) it reduces, with A β25-35
Group is compared, the EPA/DHA significantly (p of all proportions test<0.05) increase GSH contents.Protect SH-SY5Y cells from A β25-35
Induction anti-oxidant GSH reduce FAs effect be：1：2EPA/DHA≤DHA<1：1EPA/DHA<EPA≤2：1EPA/DHA.
In the present embodiment, by the SH-SY5Y cell inoculations of differentiation in six orifice plates, and addition 2mL is thin into each hole
Born of the same parents' suspension.After experiment, cell is harvested.Follow manufacturer recommendation RNA extraction method (Lipid
Tissue Handbook).Using GoScriptTM Reverse Transcriptase (a) (Promega, Canada) from RNA
Synthesize complementary DNA (cDNA).2 μ gRNA synthesize for the first chain cDNA.The nucleotide sequence of primer is from NCBI's
Nucleotide databases and Primer Premier 6.0.After specificity verification in NCBI- nucleotide-BLAST, draw
The synthesis of object is carried out by Invitrogen companies.It is prepared using Quantitect SYBR Greenmaster mix (Qiagen)
PCR reacts, and carries out PCR reactions using Real Time PCR Detection System (Bio-Rad, the U.S.) CFX96TM real-time systems.PCR processes
It is as follows：95 DEG C are initially incubated 5 minutes to activate Hot-Star-Taq archaeal dna polymerases, then 94 DEG C 15 seconds (denaturation), 59 DEG C 30
Second (annealing) and 72 DEG C 30 seconds (extensions).After 38 cycles, a melting curve is produced, for measuring the spy of primer
The opposite sex and homogeneity.The rna expression (relative quantification) of gene expression dose house-keeping gene beta-actin is corrected with △ △ CT
Standardization.
In the present embodiment, by SH-SY5Y cell inoculations in 6 orifice plates, and 2mL cells is added in into each hole and are suspended
Liquid.After experiment process, cell is collected, and centrifuge 10 minutes, and with RIPA buffer solutions (RIPA, Thermo with 10.000g
Scientific it) cracks.By ultrasonic wave Assisted Cleavage, lysate is centrifuged 10 minutes at 4 DEG C with 10000g.Collect supernatant,
After boiling add in the aliquot containing 20-40 μ g proteins, and on 10%SDS-PAGE gels with 100V in running buffer
It is detached 60 minutes in liquid.Run gel after, by Protein transfer to polyvinylidene fluoride (PVDF) film (Millipore,
Bellerica, MA, the U.S.) on.Then trace is washed 5 minutes in Tris buffer solutions-polysorbas20 (TBST), then 20
(TBST and 5% alipoidic milk power) is closed at DEG C 1 hour.It after closing, washs trace 5 minutes with TBST, is incubated with primary antibody, wrap
It includes for more grams of the rabbit source of actin, NGF, BDNF, TrkA, TrkB, TNF-α, Bcl-2, Bax and Caspase-3 (Abcam)
Grand antibody, 4 DEG C overnight, and secondary antibody, the anti-rabbit IgG of peroxidase (HRP) combination 1 hour are then added at 20 DEG C.By trace
It is washed three times in TBS.Use ClarityTMWestern ECL substrate reagent box (Bio-rad, Canada) are with Image
LabTMThe ChemiDoc of software (Bio-rad, Canada)TMImmunoreactivity band is detected in MP systems.By being normalized
For the beta-actin detected again on same film, then with the percentage calculation of control group, all target proteins are quantified.
Compared with the control group, after 4 hours are incubated, by giving A β25-35, TNF-α mRNA expression dramatically increase (p<
0.05, Fig. 4 a), but with A β25-35Increase (the P of protein expression can not be found before being incubated 12 hours<0.01).However,
With A β25-35Group is compared, and has individually been restored with the EPA and DHA being used in combination to A β25-35The TNF-α mRNA expression of processing reaction
It significantly reduces.(Fig. 4 a) is in addition, 1：The effect of 1EPA/DHA is with obvious effects more more effective than individual EPA or DHA, these evidences
Show 1:1EPA/DHA can play anti-inflammatory agent potential synergistic effect.
It is similar to TNF-α gene, A β25-35Processing occurs apparent ngf gene expression variation for 4 hours.Compared with the control group,
Aβ25-35Damaging cells NGF mRNA expression is apparent to lower (p<0.01, Fig. 5 a).Meanwhile A β25-35Damaging cells BDNF genes
(p was also lowered in mRNA expression at 4 hours<0.01, Fig. 5 b).Compared with the control, A β25-35In damaging cells, when being incubated within 12 hours
It was found that the protein expression of NGF significantly reduces (p<0.01, Fig. 5 c), and bdnf protein expression dramatically increases (p<0.01, Fig. 5 d).With
EPA, DHA and its pretreatment of various ratios can weaken A β to some extent25-35NGF mRNA and the protein expression (figure of induction
5a, c), BDNF mRNA expression (Fig. 5 b) and bdnf protein expression variation (Fig. 5 d).
Different from the gene of front, TrkA and TrkB are giving A β25-35It is affected after 24 hours.Aβ25-35It is substantially reduced
TrkA protein expressions (p<0.01).EPA, DHA and its processing of various ratios cannot significantly change A β25-35Effect to receptor, is removed
2：1EPA/DHA significantly increases this variation (Fig. 5 a).Individually giving A β25-35Cell in, it has been found that TrkB protein
Distant increase (p<0.05).2：1 and 1：2EPA/DHA pretreatments can part significantly reverse A β25-35The TrkB expression of induction
Change (equal p<0.05, Fig. 5 b), compared with A β groups, other groups handled with EPA and/or DHA are without significant change.
In A β25-35Different incubation times (4,8,12 and 24 hours) under test cdna Bax, Bcl-2 and Caspase-3 egg
White expression.Bcl-2 is with A β25-35When being incubated 24 hours, protein expression is by A β25-35It is strong to reduce, by using EPA, DHA and its
The pretreatment of various ratios dramatically increases (p in various degree<0.01).About Bax, do not found after being incubated at 4~24 hours apparent
Variation.Bax in cell：The ratio of Bcl-2 is individually giving A β25-35Dramatically increase (p<0.01).However, EPA, DHA and its
The processing of various ratios can differently be obviously reduced this effect, and by Bax：Bcl-2 ratios are reduced to different degrees of control
Level is (in addition to EPA p<0.05, all p<0.01, Fig. 6 a)
For caspase-3 mRNA, A β25-35Dramatically increase its protein expression (p<0.01), EPA, DHA and its various ratios
Example processing can weaken A β to some extent25-35Effect (in addition to 1：1EPA/DHAp<0.05, all p<0.01, Fig. 6 b).
To sum up, 1：2EPA/DHA shows most effective to cell viability reduction；2：1EPA/DHA is sent out in test group
Wave the most effective ratio of antioxidation.For antiphlogistic effects, 1：1EPA/DHA is the optimal proportion in all test ratios；
When being related to Anti-G value, pure DHA is more more effective than the combination of EPA and any other ratio.Based on these as a result, the knot obtained
By for EPA, DHA and its various ratios differently adjust A in SH-SY5Y cells by differently inhibiting inflammation and oxidative stress
β25-35The neurotoxicity of induction adjusts neurotrophic factor level, so as to weaken Neuron Apoptosis.
Claims (4)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711468538.5A CN108159037A (en) | 2017-12-29 | 2017-12-29 | A kind of unsaturated fatty-acid compositions and its application for being used to improve neuroprotective function |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711468538.5A CN108159037A (en) | 2017-12-29 | 2017-12-29 | A kind of unsaturated fatty-acid compositions and its application for being used to improve neuroprotective function |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN108159037A true CN108159037A (en) | 2018-06-15 |
Family
ID=62519793
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711468538.5A Pending CN108159037A (en) | 2017-12-29 | 2017-12-29 | A kind of unsaturated fatty-acid compositions and its application for being used to improve neuroprotective function |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN108159037A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108721269A (en) * | 2018-07-10 | 2018-11-02 | 广东海洋大学 | A kind of application of EPA with DHA intermixtures in terms of prevention anxiety and anhedonia |
| CN109045004A (en) * | 2018-07-10 | 2018-12-21 | 广东海洋大学 | A kind of EPA and DHA intermixture for preventing and treating anxiety and anhedonia |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1320939A (en) * | 2000-04-27 | 2001-11-07 | 尹顿公司 | Mechanically interlocking device for coordinating operation of circuit protectors and with overregulation compensation |
| CN106692973A (en) * | 2015-11-13 | 2017-05-24 | 深圳君圣泰生物技术有限公司 | Pharmaceutical compositions and applications thereof, and pharmaceutical preparation |
-
2017
- 2017-12-29 CN CN201711468538.5A patent/CN108159037A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1320939A (en) * | 2000-04-27 | 2001-11-07 | 尹顿公司 | Mechanically interlocking device for coordinating operation of circuit protectors and with overregulation compensation |
| CN106692973A (en) * | 2015-11-13 | 2017-05-24 | 深圳君圣泰生物技术有限公司 | Pharmaceutical compositions and applications thereof, and pharmaceutical preparation |
Non-Patent Citations (1)
| Title |
|---|
| NICOLE PAGES 等: "Brain protection by rapeseed oil in magnesium-deficient mice", 《PROSTAGLANDINS LEUKOT ESSENT FATTY ACIDS.》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108721269A (en) * | 2018-07-10 | 2018-11-02 | 广东海洋大学 | A kind of application of EPA with DHA intermixtures in terms of prevention anxiety and anhedonia |
| CN109045004A (en) * | 2018-07-10 | 2018-12-21 | 广东海洋大学 | A kind of EPA and DHA intermixture for preventing and treating anxiety and anhedonia |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| YUN-SOO et al. | Quercetin prevents adipogenesis by regulation of transcriptional factors and lipases in OP9 cells | |
| Tan et al. | Melatonin: both a messenger of darkness and a participant in the cellular actions of non-visible solar radiation of near infrared light | |
| CN1893909A (en) | Composition for improving skin, hair and coat health containing flavanones | |
| JP6403039B2 (en) | Activator of SIRT3 and SIRT6 | |
| CN101663029A (en) | Use of hydroxytyrosol as anti-aging agent | |
| KR102091620B1 (en) | Pharmaceutical composition for prevention or treatment of blood brain barrier disorder comprising l-serine as an effective component and health functional food comprising the same | |
| KR20200037731A (en) | A composition for inhibiting senescence comprising Akkermansia muciniphila or culture of the same | |
| CN108159037A (en) | A kind of unsaturated fatty-acid compositions and its application for being used to improve neuroprotective function | |
| JP7032855B2 (en) | Compositions for altering the expression of clock genes | |
| US20100099918A1 (en) | Tranquilizer and Functional Food | |
| CN111818812B (en) | Composition for improving skin condition | |
| CN108463234A (en) | The glucose metabolism activator of astroglia | |
| CN105960244A (en) | Antibacterial peptide-inducing agent | |
| KR20210012915A (en) | A pharmaceutical composition for preventing or treating inflammatory bone joint diseases comprising an enzyme-treated royal jelly powder | |
| JP6557893B1 (en) | Composition for prevention, reduction and / or treatment of heat stroke | |
| CN104825504A (en) | Anti-inflammatory application of pomegranate flower polyphenol | |
| US8367060B2 (en) | Pharmaceutical composition containing arazyme for the prevention of liver dysfunction | |
| Zinina et al. | Protein hydrolysate as a source of bioactive peptides in diabetic food products | |
| EP3878457A1 (en) | Brown/beige adipocyte activating agent containing d-allulose as active ingredient | |
| WO2015028456A1 (en) | PPAR modulators | |
| NAHYA | The Bee Bread as Super Food: An up-to-date Review in terms of Chronic Diseases | |
| CN109758444B (en) | Antioxidant application of 2-methyl substituted fatty acids | |
| Ahmad et al. | Efek Pemberian Minyak Buah Zaitun (Olea Europaea L) Dosis Rendah Terhadap Kadar Malondialdehid (MDA) Tikus Putih (Rattus Norvegicus) Obesitas | |
| Gabbianelli | 4th European Summer School on Nutrigenomics (ESSN 2021) | |
| Gabbianelli | Proceedings of the 4th European Summer School on Nutrigenomics (ESSN 2021), June 21-25, 2021 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20180615 |