CN114621319A - Probiotic active peptide, active peptide composition and application thereof in preparation of products with anti-inflammatory and/or antioxidant effects - Google Patents
Probiotic active peptide, active peptide composition and application thereof in preparation of products with anti-inflammatory and/or antioxidant effects Download PDFInfo
- Publication number
- CN114621319A CN114621319A CN202111577364.2A CN202111577364A CN114621319A CN 114621319 A CN114621319 A CN 114621319A CN 202111577364 A CN202111577364 A CN 202111577364A CN 114621319 A CN114621319 A CN 114621319A
- Authority
- CN
- China
- Prior art keywords
- active peptide
- probiotic active
- formula
- probiotic
- inflammatory
- 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
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 83
- 239000006041 probiotic Substances 0.000 title claims abstract description 76
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 76
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 75
- 239000000203 mixture Substances 0.000 title claims abstract description 46
- 230000003110 anti-inflammatory effect Effects 0.000 title claims abstract description 16
- 230000003078 antioxidant effect Effects 0.000 title claims abstract description 12
- 238000002360 preparation method Methods 0.000 title abstract description 5
- 239000003814 drug Substances 0.000 claims abstract description 17
- 239000003344 environmental pollutant Substances 0.000 claims abstract description 14
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 12
- 230000004792 oxidative damage Effects 0.000 claims abstract description 10
- 235000013305 food Nutrition 0.000 claims description 18
- 235000015872 dietary supplement Nutrition 0.000 claims description 15
- 230000006749 inflammatory damage Effects 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 230000000975 bioactive effect Effects 0.000 claims description 4
- 229940121363 anti-inflammatory agent Drugs 0.000 claims description 3
- 239000002260 anti-inflammatory agent Substances 0.000 claims description 3
- 230000003647 oxidation Effects 0.000 claims description 2
- 238000007254 oxidation reaction Methods 0.000 claims description 2
- FMMWHPNWAFZXNH-UHFFFAOYSA-N Benz[a]pyrene Chemical compound C1=C2C3=CC=CC=C3C=C(C=C3)C2=C2C3=CC=CC2=C1 FMMWHPNWAFZXNH-UHFFFAOYSA-N 0.000 abstract description 53
- 230000000694 effects Effects 0.000 abstract description 14
- 230000002757 inflammatory effect Effects 0.000 abstract description 12
- 230000006378 damage Effects 0.000 abstract description 9
- 208000027418 Wounds and injury Diseases 0.000 abstract description 6
- 208000014674 injury Diseases 0.000 abstract description 6
- 230000000968 intestinal effect Effects 0.000 abstract description 6
- 229940079593 drug Drugs 0.000 abstract description 5
- 230000003064 anti-oxidating effect Effects 0.000 abstract description 3
- 150000002500 ions Chemical class 0.000 description 22
- 210000004027 cell Anatomy 0.000 description 15
- 239000003642 reactive oxygen metabolite Substances 0.000 description 10
- 230000014509 gene expression Effects 0.000 description 8
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 description 8
- 108010002616 Interleukin-5 Proteins 0.000 description 7
- 102100029438 Nitric oxide synthase, inducible Human genes 0.000 description 7
- 101710089543 Nitric oxide synthase, inducible Proteins 0.000 description 7
- 102100038280 Prostaglandin G/H synthase 2 Human genes 0.000 description 7
- 108050003267 Prostaglandin G/H synthase 2 Proteins 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 108090000695 Cytokines Proteins 0.000 description 6
- 102000004127 Cytokines Human genes 0.000 description 6
- 108090000176 Interleukin-13 Proteins 0.000 description 6
- 108090000978 Interleukin-4 Proteins 0.000 description 6
- 230000028327 secretion Effects 0.000 description 6
- 239000003963 antioxidant agent Substances 0.000 description 5
- XEYBRNLFEZDVAW-ARSRFYASSA-N dinoprostone Chemical compound CCCCC[C@H](O)\C=C\[C@H]1[C@H](O)CC(=O)[C@@H]1C\C=C/CCCC(O)=O XEYBRNLFEZDVAW-ARSRFYASSA-N 0.000 description 5
- 229960002986 dinoprostone Drugs 0.000 description 5
- XEYBRNLFEZDVAW-UHFFFAOYSA-N prostaglandin E2 Natural products CCCCCC(O)C=CC1C(O)CC(=O)C1CC=CCCCC(O)=O XEYBRNLFEZDVAW-UHFFFAOYSA-N 0.000 description 5
- TXVHTIQJNYSSKO-UHFFFAOYSA-N benzo[e]pyrene Chemical class C1=CC=C2C3=CC=CC=C3C3=CC=CC4=CC=C1C2=C34 TXVHTIQJNYSSKO-UHFFFAOYSA-N 0.000 description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 4
- 230000003833 cell viability Effects 0.000 description 4
- 239000002609 medium Substances 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 3
- 239000000356 contaminant Substances 0.000 description 3
- 230000007613 environmental effect Effects 0.000 description 3
- 238000004128 high performance liquid chromatography Methods 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 230000003834 intracellular effect Effects 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 241000218588 Lactobacillus rhamnosus Species 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000001035 drying Methods 0.000 description 2
- 238000002101 electrospray ionisation tandem mass spectrometry Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- VNWKTOKETHGBQD-UHFFFAOYSA-N methane Chemical compound C VNWKTOKETHGBQD-UHFFFAOYSA-N 0.000 description 2
- 125000001997 phenyl group Chemical group [H]C1=C([H])C([H])=C(*)C([H])=C1[H] 0.000 description 2
- 230000001681 protective effect Effects 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- BBEAQIROQSPTKN-UHFFFAOYSA-N pyrene Chemical compound C1=CC=C2C=CC3=CC=CC4=CC=C1C2=C43 BBEAQIROQSPTKN-UHFFFAOYSA-N 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- 230000035899 viability Effects 0.000 description 2
- VQVUBYASAICPFU-UHFFFAOYSA-N (6'-acetyloxy-2',7'-dichloro-3-oxospiro[2-benzofuran-1,9'-xanthene]-3'-yl) acetate Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Cl)=C(OC(C)=O)C=C1OC1=C2C=C(Cl)C(OC(=O)C)=C1 VQVUBYASAICPFU-UHFFFAOYSA-N 0.000 description 1
- XXMFJKNOJSDQBM-UHFFFAOYSA-N 2,2,2-trifluoroacetic acid;hydrate Chemical compound [OH3+].[O-]C(=O)C(F)(F)F XXMFJKNOJSDQBM-UHFFFAOYSA-N 0.000 description 1
- 102400000888 Cholecystokinin-8 Human genes 0.000 description 1
- 101800005151 Cholecystokinin-8 Proteins 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 238000008157 ELISA kit Methods 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102100038277 Prostaglandin G/H synthase 1 Human genes 0.000 description 1
- 108050003243 Prostaglandin G/H synthase 1 Proteins 0.000 description 1
- 230000006052 T cell proliferation Effects 0.000 description 1
- 210000001744 T-lymphocyte Anatomy 0.000 description 1
- 206010043275 Teratogenicity Diseases 0.000 description 1
- 210000004241 Th2 cell Anatomy 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000009285 allergic inflammation Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 150000004945 aromatic hydrocarbons Chemical class 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000002238 attenuated effect Effects 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 230000000711 cancerogenic effect Effects 0.000 description 1
- 231100000357 carcinogen Toxicity 0.000 description 1
- 239000003183 carcinogenic agent Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 208000037893 chronic inflammatory disorder Diseases 0.000 description 1
- 239000003245 coal Substances 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000003828 downregulation Effects 0.000 description 1
- 238000002330 electrospray ionisation mass spectrometry Methods 0.000 description 1
- 210000002889 endothelial cell Anatomy 0.000 description 1
- 210000003979 eosinophil Anatomy 0.000 description 1
- GVEPBJHOBDJJJI-UHFFFAOYSA-N fluoranthrene Natural products C1=CC(C2=CC=CC=C22)=C3C2=CC=CC3=C1 GVEPBJHOBDJJJI-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 239000000446 fuel Substances 0.000 description 1
- 210000001035 gastrointestinal tract Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 210000003630 histaminocyte Anatomy 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000004957 immunoregulator effect Effects 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000006166 lysate Substances 0.000 description 1
- UHNSRFWQBVXBSK-UHFFFAOYSA-N methanol;2,2,2-trifluoroacetic acid Chemical compound OC.OC(=O)C(F)(F)F UHNSRFWQBVXBSK-UHFFFAOYSA-N 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000007886 mutagenicity Effects 0.000 description 1
- 231100000299 mutagenicity Toxicity 0.000 description 1
- 239000003345 natural gas Substances 0.000 description 1
- 230000008520 organization Effects 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 238000012261 overproduction Methods 0.000 description 1
- 230000001590 oxidative effect Effects 0.000 description 1
- 230000036542 oxidative stress Effects 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 210000004180 plasmocyte Anatomy 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000002953 preparative HPLC Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000031337 regulation of inflammatory response Effects 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 230000009528 severe injury Effects 0.000 description 1
- IZTQOLKUZKXIRV-YRVFCXMDSA-N sincalide Chemical compound C([C@@H](C(=O)N[C@@H](CCSC)C(=O)NCC(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCSC)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC=1C=CC=CC=1)C(N)=O)NC(=O)[C@@H](N)CC(O)=O)C1=CC=C(OS(O)(=O)=O)C=C1 IZTQOLKUZKXIRV-YRVFCXMDSA-N 0.000 description 1
- 230000000391 smoking effect Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 231100000211 teratogenicity Toxicity 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000004227 thermal cracking Methods 0.000 description 1
- 238000001196 time-of-flight mass spectrum Methods 0.000 description 1
- 230000002103 transcriptional effect Effects 0.000 description 1
- 239000002912 waste gas Substances 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/12—Cyclic peptides with only normal peptide bonds in the ring
- C07K5/126—Tetrapeptides
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- 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
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Pharmacology & Pharmacy (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Food Science & Technology (AREA)
- Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Mycology (AREA)
- Genetics & Genomics (AREA)
- Biophysics (AREA)
- Rheumatology (AREA)
- Pain & Pain Management (AREA)
- Toxicology (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
The invention relates to the technical field of biological medicines, and particularly discloses a probiotic active peptide, a probiotic active peptide composition and application of the probiotic active peptide and the probiotic active peptide composition in preparation of products with anti-inflammatory and/or anti-oxidation effects. The probiotic active peptide has a structure shown in a formula I or a formula II. The probiotic active peptide composition comprises probiotic active peptides with structures shown in a formula I and a formula II. The probiotic active peptide and the composition thereof have anti-inflammatory and antioxidant effects; especially has the effect of resisting inflammatory injury and oxidative injury of intestinal cells caused by benzopyrene as an environmental pollutant.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to a probiotic active peptide, a probiotic active peptide composition and application of the probiotic active peptide and the probiotic active peptide composition in preparation of products with anti-inflammatory and/or anti-oxidation effects.
Background
Benzopyrene (abbreviated as BaP), which is a condensed ring aromatic hydrocarbon containing a benzene ring; benzopyrenes have various isomers according to the fused position of a benzene ring, and benzo [ a ] pyrene and benzo [ e ] pyrene are two common benzopyrenes; is a class I carcinogen determined by the world health organization, and has extremely strong teratogenicity and mutagenicity.
The food can generate thermal cracking reaction under high-temperature processing conditions such as frying, smoking, baking and the like, and then the benzopyrene is formed through reactions such as cyclization, polymerization and the like; in addition, in industrial production and life, fuels such as coal, petroleum, natural gas and the like are not completely combusted to generate BaP-containing waste gas, so that food is polluted by water sources, air, soil and the like.
However, a great deal of research finds that the BaP in the food has induction and promotion effects on the occurrence and development of various digestive tract diseases. However, there is currently no therapeutic product for oxidative damage and inflammatory damage caused by benzopyrene, which is an environmental pollutant.
Disclosure of Invention
In order to overcome at least one of the technical problems of the prior art, the present invention firstly provides a probiotic active peptide.
The detailed technical scheme of the invention is as follows:
the invention firstly provides a probiotic active peptide, which has a structure shown in a formula I or a formula II:
the inventor surprisingly found in the research that: the probiotic active peptide with the structure shown in the formula I or the formula II has excellent antioxidant and anti-inflammatory effects; especially has excellent effects of resisting environmental pollutants and oxidizing damage and inflammatory damage caused by pyrene.
Wherein, the probiotic active peptide with the structure shown in the formula I is cyclo [ Thr-His-Ala-Trp ] peptide, which is abbreviated as LRCP-1 in the following; the probiotic active peptide with the structure shown in the formula II is cyclo [ His-Phe-Leu-Val ] peptide, which is abbreviated as LRCP-2 in the following.
The invention also provides a probiotic active peptide composition, which comprises probiotic active peptides with structures shown in formula I and formula II.
Furthermore, the research of the inventor shows that after the probiotic active peptide with the structures shown in the formula I and the formula II is combined, the antioxidant and anti-inflammatory effects, especially the effects of resisting oxidative damage and inflammatory damage caused by environmental pollutant benzopyrene, of the probiotic active peptide are better than that of the probiotic active peptide with the structure shown in the formula I.
Preferably, the molar ratio of the probiotic active peptide with the structures shown in the formula I and the formula II is 1-10: 1-10.
Further preferably, the molar ratio of the probiotic active peptide with the structures shown in the formula I and the formula II is 1-5: 1-5.
Most preferably, the molar ratio of the probiotic active peptides having the structures shown in formula I and formula II is 1: 1.
The invention also provides application of the probiotic active peptide or the probiotic active peptide composition in preparation of food, dietary supplements or medicines.
Preferably, the food, dietary supplement or medicament is a food, dietary supplement or medicament having an antioxidant effect.
Preferably, the oxidation resistance is resistance to oxidative damage caused by environmental contaminants.
Most preferably, the environmental contaminant is benzopyrene.
Most preferably, the oxidative damage is oxidative damage of intestinal cells.
The probiotic active peptide and the probiotic active peptide composition have excellent antioxidation, and especially have excellent effect of resisting oxidative damage caused by environmental pollutant benzopyrene; therefore, the compound can be used as an active ingredient for preparing foods, dietary supplements or medicaments with an antioxidant effect, particularly an effect of resisting oxidative damage caused by environmental pollutants benzopyrene.
Preferably, the food, dietary supplement or medicament is a food, dietary supplement or medicament with anti-inflammatory effect.
Further preferably, the anti-inflammatory agent is against inflammatory damage caused by environmental pollutants.
Most preferably, the environmental contaminant is benzopyrene.
Most preferably, the inflammatory injury is inflammatory injury of intestinal cells.
The probiotic active peptide and the probiotic active peptide composition have excellent anti-inflammatory effect, and especially have excellent effect of resisting inflammatory injury caused by environmental pollutant benzopyrene; therefore, the compound can be used as an active ingredient for preparing foods, dietary supplements or medicaments with anti-inflammatory effect, particularly for preparing foods, dietary supplements or medicaments with the effect of resisting inflammatory injury caused by environmental pollutants benzopyrene.
Has the advantages that: (1) the invention provides probiotic active peptides and compositions thereof. Experimental results show that the probiotic active peptide and the composition thereof can inhibit excessive secretion of intestinal inflammatory factors and excessive production of ROS (reactive oxygen species) caused by benzopyrene serving as an environmental pollutant. This shows that the probiotic active peptide and the composition thereof have excellent anti-inflammatory and antioxidant functions; especially can inhibit inflammatory injury and oxidative injury of intestinal endothelial cells caused by benzopyrene as an environmental pollutant. Further research shows that the probiotic active peptide composition has more obvious inhibiting effect on excessive secretion of intestinal cell inflammatory factors and excessive generation of ROS (reactive oxygen species) caused by benzopyrene serving as an environmental pollutant, and shows that the mixed probiotic active peptides play a synergistic anti-inflammatory and antioxidant function.
(2) The probiotic active peptide and the composition thereof have excellent anti-inflammatory and antioxidant activities. Therefore, the probiotic active peptide and the composition thereof can be used as anti-inflammatory and antioxidant active substances and have wide application prospects in foods, dietary supplements or medicines.
(3) The probiotic active peptide can be obtained by separating from lactobacillus rhamnosus lysate, has rich sources, and is beneficial to application in food, dietary supplements or medicines.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the embodiments or the prior art descriptions will be briefly described below, it is obvious that the drawings in the following description are only drawings of some embodiments of the present invention, and other drawings can be obtained by those skilled in the art without creative efforts.
FIG. 1 is a mass spectrum of LRCP-1.
FIG. 2 is an HPLC chart of LRCP-1.
FIG. 3 is a mass spectrum of LRCP-2.
FIG. 4 is an HPLC chart of LRCP-2.
FIG. 5 is a graph showing the results of experiments in which LRCP-1, LRCP-2, or a combination thereof inhibited BaP-induced ROS production.
FIG. 6 is a graph showing the results of experiments in which LRCP-1, LRCP-2, or a combination thereof inhibits BaP-induced inflammatory cytokine secretion.
FIG. 7 is a graph showing the results of experiments in which LRCP-1, LRCP-2 or a combination thereof inhibited BaP-induced secretion of NO, PGE 2.
FIG. 8 is a graph showing the results of experiments in which LRCP-1, LRCP-2, or a combination thereof inhibits the expression of COX-2 and iNOS induced by BaP.
Detailed Description
The technical solution of the present invention will be clearly and completely described with reference to the following examples. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Mixing 50g of lactobacillus rhamnosus with 300mL of water, carrying out ultrasonic crushing, and centrifuging to obtain a water layer; repeating the steps for 3 times, freeze-drying the water layer at low temperature in vacuum, and then preparing the probiotic active peptide with the structure shown in formula I (LRCP-1) and formula II (LRCP-2) by using HPLC.
Conditions for preparative HPLC were: adopting C18 reversed phase preparative chromatographic column; taking 0.1% trifluoroacetic acid water solution as mobile phase A, taking 0.1% trifluoroacetic acid methanol solution as mobile phase A, wherein the ratio of mobile phase A: mobile phase B60: 40; the measuring wavelength is 310 μm; collecting the chromatographic peak of 12.49min and the fraction corresponding to the chromatographic peak of 12.16min, concentrating and drying to obtain LRCP-1 and LRCP-2.
The prepared LRCP-1 and LRCP-2 were subjected to ESI-MS analysis using a SCIEX X500R Q-TOF mass spectrometer equipped with an ESI source. The MS conditions were as follows: the mass range is set to be m/z 50-1500. Obtaining Q-TOF mass spectrum data in a positive mode, wherein the mass spectrum analysis conditions are as follows: CAD gas flow, 7L/min; the temperature of the drying gas is 550 ℃; ion spraying voltage, 5500V; potential, 80 volts. Data files generated by software: SCIEX OS 1.0. The analytical results were as follows:
resolution of LRCP-1: 496.7325 is [ M + H]+(ii) a Ion 478.2213 is [ M-H2O+H]+(ii) a Ion 460.7127 is [ M-2H2O+H]+(ii) a Ion 425.1942 is [ M-Ala + H2O+H]+(ii) a Ion 258.1244 is [ Trp-Ala-2H2O+H]+(ii) a Ion 239.1140 is [ His-Thr-2H2O+H]+(ii) a Ion 209.1035 is [ Ala-His-2H2O+H]+(ii) a Ion 159.0920 is [ Trp-COOH + H]+(ii) a Ion 110.0721 is [ His-COOH + H]+(ii) a Based on ESI-MS/MS data, it was determined that LRCP-1 was cyclo [ Thr-His-Ala-Trp]Peptide, namely probiotic active peptide with the structure shown in formula I.
Resolution of LRCP-2: ion 497.2843[ M + H]+(ii) a Ion 469.2897 is [ M-CO + H]+(ii) a Ion 398.2164 is [ M-Val + H2O+H]+(ii) a The ion 384.2012 being [ M-Leu + H2O+H]+(ii) a Ion 380.2053 is [ M-Val + H]+(ii) a Ion 350.2168 is [ M-Phe + H2O+H]+(ii) a Ion 285.1325 is [ His-Phe-2H2O+H]+(ii) a Ion 261.1582 is [ Leu-Phe-2H2O+H]+(ii) a Ion 237.1332 is [ His-Val-2H2O+H]+(ii) a The ion 212.1169 being [ Val-Leu-2H2O+H]+(ii) a Ion 138.0656 is [ His-H2O+H]+(ii) a Ion 110.0708 is [ His-COOH + H]+(ii) a Ion 86.0975 is [ Leu-COOH + H]+(ii) a Based on ESI-MS/MS data, it was determined that LRCP-2 was cyclo [ His-Phe-Leu-Val ]]Peptide, namely probiotic active peptide with the structure shown in formula II.
Examples of the experiments
In order to evaluate the biological activity of the above-mentioned probiotic active peptides LRCP-1 and LRCP-2 and probiotic active peptide composition (composition consisting of LRCP-1 and LRCP-2 in a molar ratio of 1:1, abbreviated as composition) of the present invention, the following effect experimental examples were conducted. BaP in the following experimental examples was benzo [ a ] pyrene.
Caco-2 cells were seeded in 6-well plates (4X 10)5cells/mL) and stimulated with BaP (10nm) for 12 hours. Thereafter, the medium was removed, then washed twice with PBS (pH 7.2), and then the cells were cultured in new medium (LRCP-1, LRCP-2 or and probiotic bioactive peptide composition) containing the sample for 24 hours. The viability of Caco-2 cells was determined using the CCK8 assay. Viability of Caco-2 cells in each well was expressed as a percentage of control.
Caco-2 cells were seeded in 6-well plates (4X 10)5cells/mL) and stimulated with BaP (10nm) for 12 hours. The medium was removed, then washed twice with PBS (pH 7.2), and then the cells were cultured in new medium containing LRCP-1 or probiotic active peptide composition for 24 hours. Cells were harvested and stained with 30 μm 2 ', 7' -dichlorofluorescein diacetate for 30 min at 37 ℃. The cells were then washed twice with PBS and placed in a microplate reader. Meanwhile, collecting the culture medium and detecting the levels of related inflammatory factors and ROS by using an ELISA kit; the cells were collected. Extracting protein, and detecting the change condition of the protein expression of the related signal path by a western blotting method.
The results are as follows:
TABLE 1 Probiotics active peptides and compositions thereof inhibit BaP-induced cell viability decline
(n＝3,#p<0.05, compared to blank;*p<0.05, compared to BaP group).
As shown in table 1, BaP exposure reduced cell viability by 18.2% (p <0.05), indicating that BaP exposure caused severe damage to cells. However, BaP-induced reduction in cell viability was significantly reversed after treatment with the probiotic active peptides or compositions thereof. Among them, LRCP-1 showed 13.8% increase in cell viability (p <0.01) compared to the BaP-induced group, and had more effective protective activity against BaP-induced damage than LRCP-2. Furthermore, the probiotic active peptide composition has a more effective protective activity against BaP-induced damage than LRCP-1 and LRCP-2 as well. We therefore conducted more in-depth experiments with LRCP-1 and probiotic active peptide compositions.
Studies have shown that BaP exposure induces excessive intracellular ROS production, which in turn leads to oxidative stress and apoptosis. In this experiment, BaP exposure induced an increase in intracellular ROS levels of 453% (p <0.01) compared to the blank control. Both LRCP-1 and probiotic active peptide compositions (abbreviated as compositions in the figure) treated reduced intracellular ROS levels compared to the BaP group. Among them, the probiotic active peptide composition inhibited BaP-induced ROS overproduction more significantly than LRCP-1 (fig. 5).
Proinflammatory cytokines, such as IL-4, IL-5 and IL-13, play an important role in the regulation of immunity and inflammation. IL-4 has activity in modulating inflammation, stimulating mast cell and T cell proliferation. IL-5 is a T cell-derived cytokine that triggers the eventual differentiation of activated B cells into antibody-secreting plasma cells. Overexpression of IL-5 increases eosinophil number and antibody levels in vivo. IL-13 is an immunoregulatory cytokine secreted primarily by activated Th2 cells and plays a key regulatory role in the pathogenesis of allergic inflammation. We examined the effect of LRCP-1 and probiotic active peptide compositions on BaP-induced IL-4, IL-5, IL-13 and IFN- γ release by ELISA. As shown in fig. 6, IL-4, IL-5, IL-13 and IFN- γ levels were significantly up-regulated by 864.8% (p <0.01), 503.4% (p <0.01), 326.8% (p <0.01) and 644.3% (p <0.01), respectively, compared to the blank control group. However, the LRCP-1 and probiotic active peptide compositions attenuated BaP-induced excessive release of IL-4, IL-5, IL-13, and IFN- γ in Caco-2 cells. Treatment with the probiotic active peptide composition reduced BaP-induced IL-4, IL-5, IL-13 and IFN- γ expression by 44.3% (p <0.01), 54.8% (p <0.01), 45.8% (p <0.01) and 61.5% (p <0.01), respectively, as compared to the BaP group. The probiotic active peptide composition can obviously inhibit inflammatory cytokine secretion caused by BaP exposure and inhibit inflammatory damage caused by BaP exposure.
Over-secretion, expression of NO and PGE2 and synergy of NO, PGE2 with other inflammatory cytokines exacerbate inflammation. To evaluate the anti-inflammatory effects of LRCP-1 and probiotic bioactive peptide compositions, we examined the changes in NO and PGE2 levels, and the results are shown in fig. 7. BaP exposure significantly induced NO release. BaP exposure increased NO levels by 228% (p <0.01) compared to the blank control group. However, LRCP-1 and probiotic active peptide compositions reduced BaP-induced NO production by 35.7% (p <0.05) and 45.1% (p < 0.01). Similarly, LRCP-1 and probiotic active peptide compositions also inhibited BaP-induced excessive release of PGE 2.
iNOS is an enzyme involved in excess NO production during chronic inflammatory diseases. COX-1 was able to regulate the homeostasis of PGE2, while COX-2 was able to induce an excessive release of PGE 2. Therefore, iNOS and COX-2 play important roles in the regulation of inflammatory responses at the transcriptional level. We examined the effect of LRCP-1 and probiotic active peptide compositions on BaP-induced expression of iNOS and COX-2. As shown in FIG. 8, BaP significantly induced the expression of iNOS and COX-2, while LRCP-1 and probiotic active peptide compositions inhibited the expression of iNOS and COX-2. The inhibition of Caco-2 cell NO and PGE2 excessive secretion induced by BaP stimulation by LRCP-1 and probiotic active peptide composition is related to the down regulation of iNOS and COX-2 expression.
While the invention has been described in connection with what is presently considered to be the most practical and preferred embodiment, it is to be understood that the invention is not to be limited to the disclosed embodiment, but on the contrary, is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.
Claims (10)
1. A probiotic active peptide is characterized by having a structure shown as a formula I or a formula II:
2. a probiotic active peptide composition is characterized by comprising probiotic active peptides with structures shown in formula I and formula II.
3. The probiotic active peptide composition according to claim 2, characterized in that the molar ratio of the probiotic active peptides having the structures shown in formula I and formula II is 1-10: 1-10.
4. The probiotic active peptide composition of claim 3, wherein the molar ratio of the probiotic active peptides having the structures shown in formula I and formula II is 1-5: 1-5.
5. The probiotic bioactive peptide composition of claim 4, wherein the molar ratio of the probiotic bioactive peptides having the structures shown in formula I and formula II is 1: 1.
6. Use of a probiotic active peptide or probiotic active peptide composition according to any one of claims 1 to 5 in the manufacture of a food product, dietary supplement or medicament.
7. The use according to claim 6, wherein the food, dietary supplement or medicament is a food, dietary supplement or medicament having an antioxidant effect.
8. The use according to claim 7, wherein the oxidation resistance is against oxidative damage caused by environmental pollutants.
9. The use according to claim 6, wherein the food, dietary supplement or medicament is a food, dietary supplement or medicament with anti-inflammatory effect.
10. The use according to claim 9, wherein the anti-inflammatory agent is an anti-inflammatory agent against inflammatory damage caused by environmental pollutants.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111577364.2A CN114621319A (en) | 2021-12-22 | 2021-12-22 | Probiotic active peptide, active peptide composition and application thereof in preparation of products with anti-inflammatory and/or antioxidant effects |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111577364.2A CN114621319A (en) | 2021-12-22 | 2021-12-22 | Probiotic active peptide, active peptide composition and application thereof in preparation of products with anti-inflammatory and/or antioxidant effects |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114621319A true CN114621319A (en) | 2022-06-14 |
Family
ID=81898821
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111577364.2A Pending CN114621319A (en) | 2021-12-22 | 2021-12-22 | Probiotic active peptide, active peptide composition and application thereof in preparation of products with anti-inflammatory and/or antioxidant effects |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114621319A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112812150A (en) * | 2021-01-29 | 2021-05-18 | 深圳海创生物科技有限公司 | Active cyclic peptide, active cyclic peptide composition and application of active cyclic peptide composition in preparation of products with antioxidant or anti-inflammatory effects |
| CN113521248A (en) * | 2021-09-03 | 2021-10-22 | 深圳海创生物科技有限公司 | Application of probiotic active peptide in preparation of medicine, food or health-care product with effect of preventing or treating diarrhea |
-
2021
- 2021-12-22 CN CN202111577364.2A patent/CN114621319A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112812150A (en) * | 2021-01-29 | 2021-05-18 | 深圳海创生物科技有限公司 | Active cyclic peptide, active cyclic peptide composition and application of active cyclic peptide composition in preparation of products with antioxidant or anti-inflammatory effects |
| CN113521248A (en) * | 2021-09-03 | 2021-10-22 | 深圳海创生物科技有限公司 | Application of probiotic active peptide in preparation of medicine, food or health-care product with effect of preventing or treating diarrhea |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | Beneficial effects of tea water extracts on the body weight and gut microbiota in C57BL/6J mice fed with a high-fat diet | |
| Shahbazi et al. | Anti-inflammatory and immunomodulatory properties of fermented plant foods | |
| Rajapakse et al. | Purification of a radical scavenging peptide from fermented mussel sauce and its antioxidant properties | |
| De Angelis et al. | Fermentation by selected sourdough lactic acid bacteria to decrease coeliac intolerance to rye flour | |
| EP2391374B1 (en) | Composition from sphaeranthus indicus and garcinia mangostana for the control of metabolic syndrome | |
| CA2803322C (en) | Fermented soya based mixture comprising isoflavones-aglicones, equol andlunasil, process for the preparation and uses thereof in food, medical and cosmetic fields. | |
| Rizzello et al. | Lactic acid fermentation as a tool to enhance the functional features of Echinacea spp | |
| Gao et al. | Sturgeon hydrolysates alleviate DSS-induced colon colitis in mice by modulating NF-κB, MAPK, and microbiota composition | |
| CA2559596A1 (en) | Antioxidant producing bacterium and uses thereof | |
| Luo et al. | Protective effects of radish (Raphanus sativus L.) leaves extract against hydrogen peroxide-induced oxidative damage in human fetal lung fibroblast (MRC-5) cells | |
| Zhang et al. | The immunoenhancement effects of sea buckthorn pulp oil in cyclophosphamide-induced immunosuppressed mice | |
| Li et al. | Rabdosia serra alleviates dextran sulfate sodium salt-induced colitis in mice through anti-inflammation, regulating Th17/Treg balance, maintaining intestinal barrier integrity, and modulating gut microbiota | |
| Punampalam et al. | Evaluation of antioxidant properties of phycobiliproteins and phenolic compounds extracted from Bangia atropurpurea | |
| Wang et al. | Phenolics from noni (Morinda citrifolia L.) fruit alleviate obesity in high fat diet-fed mice via modulating the gut microbiota and mitigating intestinal damage | |
| Jeampakdee et al. | The apoptotic and free radical–scavenging abilities of the protein hydrolysate obtained from chicken feather meal | |
| Medjeber et al. | Ex vivo immunomodulatory effect of ethanolic extract of propolis during Celiac Disease: Involvement of nitric oxide pathway | |
| Huang et al. | Red yeast rice dietary intervention reduces oxidative stress-related inflammation and improves intestinal microbiota | |
| CN114621319A (en) | Probiotic active peptide, active peptide composition and application thereof in preparation of products with anti-inflammatory and/or antioxidant effects | |
| Iga-Buitrón et al. | Lactic fermentation of broccoli (Brassica oleracea var. italica) to enhance the antioxidant and antiproliferative activities | |
| Correa et al. | Bioavailability of plant pigment phytochemicals in Angelica keiskei in older adults: A pilot absorption kinetic study | |
| CN113164533A (en) | Probiotic supplement for metabolic health comprising lactobacillus | |
| Nagai et al. | Antioxidative activity of water extracts from the yam (Dioscorea opposita Thunb.) tuber mucilage tororo | |
| Arellano-García et al. | Lactobacillus rhamnosus GG administration partially prevents diet-induced insulin resistance in rats: a comparison with its heat-inactivated parabiotic | |
| Choi et al. | Pharmacological effects of fermented red pepper | |
| KR20230037245A (en) | Composition for preventing, improving, or treating inflammatory disease containing Lactobacillus rhamnosus PL60 as an active ingredient |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |