CN110835614B - 乳双歧杆菌gkk2、含其的组合物及其改善过敏性气喘的用途 - Google Patents
乳双歧杆菌gkk2、含其的组合物及其改善过敏性气喘的用途 Download PDFInfo
- Publication number
- CN110835614B CN110835614B CN201811083035.0A CN201811083035A CN110835614B CN 110835614 B CN110835614 B CN 110835614B CN 201811083035 A CN201811083035 A CN 201811083035A CN 110835614 B CN110835614 B CN 110835614B
- Authority
- CN
- China
- Prior art keywords
- bifidobacterium lactis
- gkk2
- composition
- allergic asthma
- subject
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 229940009289 bifidobacterium lactis Drugs 0.000 title claims abstract description 53
- 241000901050 Bifidobacterium animalis subsp. lactis Species 0.000 title claims abstract description 50
- 239000000203 mixture Substances 0.000 title claims abstract description 40
- 208000006673 asthma Diseases 0.000 title claims abstract description 38
- 201000009961 allergic asthma Diseases 0.000 title claims abstract description 30
- 210000002966 serum Anatomy 0.000 claims abstract description 33
- 239000007788 liquid Substances 0.000 claims abstract description 19
- 239000001963 growth medium Substances 0.000 claims abstract description 13
- 108010063907 Glutathione Reductase Proteins 0.000 claims abstract description 12
- 102100036442 Glutathione reductase, mitochondrial Human genes 0.000 claims abstract description 12
- 230000000241 respiratory effect Effects 0.000 claims abstract description 11
- 239000012530 fluid Substances 0.000 claims abstract description 4
- 210000005228 liver tissue Anatomy 0.000 claims abstract description 4
- 241000894006 Bacteria Species 0.000 claims description 24
- 230000001580 bacterial effect Effects 0.000 claims description 14
- 235000013305 food Nutrition 0.000 claims description 12
- 238000009630 liquid culture Methods 0.000 claims description 12
- 241000186660 Lactobacillus Species 0.000 claims description 11
- 229940039696 lactobacillus Drugs 0.000 claims description 11
- 238000004321 preservation Methods 0.000 claims description 11
- 239000013543 active substance Substances 0.000 claims description 10
- 230000006872 improvement Effects 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 9
- 239000003085 diluting agent Substances 0.000 claims description 7
- 239000000843 powder Substances 0.000 claims description 7
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 6
- 239000000654 additive Substances 0.000 claims description 6
- 238000004108 freeze drying Methods 0.000 claims description 6
- 239000003755 preservative agent Substances 0.000 claims description 6
- 206010006482 Bronchospasm Diseases 0.000 claims description 5
- 230000007885 bronchoconstriction Effects 0.000 claims description 5
- 239000007921 spray Substances 0.000 claims description 5
- 230000000996 additive effect Effects 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 4
- 235000003599 food sweetener Nutrition 0.000 claims description 4
- 244000005700 microbiome Species 0.000 claims description 4
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 4
- 239000010802 sludge Substances 0.000 claims description 4
- 239000003765 sweetening agent Substances 0.000 claims description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 3
- 241001052560 Thallis Species 0.000 claims description 3
- -1 absorption promoters Substances 0.000 claims description 3
- 238000005273 aeration Methods 0.000 claims description 3
- 238000004378 air conditioning Methods 0.000 claims description 3
- 235000013361 beverage Nutrition 0.000 claims description 3
- 239000001569 carbon dioxide Substances 0.000 claims description 3
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 3
- 235000013365 dairy product Nutrition 0.000 claims description 3
- 235000015872 dietary supplement Nutrition 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 235000013402 health food Nutrition 0.000 claims description 3
- 239000003094 microcapsule Substances 0.000 claims description 3
- JCXJVPUVTGWSNB-UHFFFAOYSA-N nitrogen dioxide Inorganic materials O=[N]=O JCXJVPUVTGWSNB-UHFFFAOYSA-N 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 230000009467 reduction Effects 0.000 claims description 3
- 239000000829 suppository Substances 0.000 claims description 3
- 229940124532 absorption promoter Drugs 0.000 claims description 2
- 239000003708 ampul Substances 0.000 claims description 2
- 239000007937 lozenge Substances 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims description 2
- 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 abstract description 26
- 239000002609 medium Substances 0.000 abstract description 16
- 229960003180 glutathione Drugs 0.000 abstract description 13
- 239000003833 bile salt Substances 0.000 abstract description 11
- 108090000854 Oxidoreductases Proteins 0.000 abstract description 5
- 238000010438 heat treatment Methods 0.000 abstract description 4
- 230000005757 colony formation Effects 0.000 abstract description 3
- 229940093761 bile salts Drugs 0.000 abstract description 2
- 230000008602 contraction Effects 0.000 abstract 1
- 210000003437 trachea Anatomy 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 36
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 26
- 239000013566 allergen Substances 0.000 description 20
- 238000012360 testing method Methods 0.000 description 20
- 241001134770 Bifidobacterium animalis Species 0.000 description 18
- 239000000523 sample Substances 0.000 description 16
- 239000004310 lactic acid Substances 0.000 description 13
- 235000014655 lactic acid Nutrition 0.000 description 13
- 239000000243 solution Substances 0.000 description 13
- 108010058846 Ovalbumin Proteins 0.000 description 12
- 210000004027 cell Anatomy 0.000 description 12
- 238000006243 chemical reaction Methods 0.000 description 12
- 229940092253 ovalbumin Drugs 0.000 description 12
- 238000002474 experimental method Methods 0.000 description 11
- 238000005406 washing Methods 0.000 description 11
- 108060003951 Immunoglobulin Proteins 0.000 description 10
- 241001465754 Metazoa Species 0.000 description 10
- 241000699666 Mus <mouse, genus> Species 0.000 description 10
- 102000018358 immunoglobulin Human genes 0.000 description 10
- 230000002829 reductive effect Effects 0.000 description 10
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 239000000427 antigen Substances 0.000 description 9
- 108091007433 antigens Proteins 0.000 description 9
- 102000036639 antigens Human genes 0.000 description 9
- 229940118852 bifidobacterium animalis Drugs 0.000 description 9
- 210000004369 blood Anatomy 0.000 description 9
- 239000008280 blood Substances 0.000 description 9
- 241000186000 Bifidobacterium Species 0.000 description 8
- 206010020751 Hypersensitivity Diseases 0.000 description 8
- 239000011534 wash buffer Substances 0.000 description 8
- 230000000172 allergic effect Effects 0.000 description 7
- 208000010668 atopic eczema Diseases 0.000 description 7
- WVDDGKGOMKODPV-UHFFFAOYSA-N Benzyl alcohol Chemical compound OCC1=CC=CC=C1 WVDDGKGOMKODPV-UHFFFAOYSA-N 0.000 description 6
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical compound CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 6
- 208000026935 allergic disease Diseases 0.000 description 6
- 230000007815 allergy Effects 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 6
- 230000000694 effects Effects 0.000 description 6
- 208000024891 symptom Diseases 0.000 description 6
- 239000002253 acid Substances 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- NZWOPGCLSHLLPA-UHFFFAOYSA-N methacholine Chemical compound C[N+](C)(C)CC(C)OC(C)=O NZWOPGCLSHLLPA-UHFFFAOYSA-N 0.000 description 5
- 229960002329 methacholine Drugs 0.000 description 5
- 238000009629 microbiological culture Methods 0.000 description 5
- 239000002504 physiological saline solution Substances 0.000 description 5
- 239000006041 probiotic Substances 0.000 description 5
- 235000018291 probiotics Nutrition 0.000 description 5
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 108010024636 Glutathione Proteins 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 206010070834 Sensitisation Diseases 0.000 description 4
- 102000005686 Serum Globulins Human genes 0.000 description 4
- 230000000903 blocking effect Effects 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 235000011187 glycerol Nutrition 0.000 description 4
- 210000003630 histaminocyte Anatomy 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 210000002345 respiratory system Anatomy 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 230000008313 sensitization Effects 0.000 description 4
- 239000007787 solid Substances 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 238000002965 ELISA Methods 0.000 description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 3
- 239000002671 adjuvant Substances 0.000 description 3
- 238000010171 animal model Methods 0.000 description 3
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N dimethyl sulfoxide Natural products CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 3
- 210000000987 immune system Anatomy 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 230000002335 preservative effect Effects 0.000 description 3
- 230000000529 probiotic effect Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 238000000926 separation method Methods 0.000 description 3
- UAIUNKRWKOVEES-UHFFFAOYSA-N 3,3',5,5'-tetramethylbenzidine Chemical compound CC1=C(N)C(C)=CC(C=2C=C(C)C(N)=C(C)C=2)=C1 UAIUNKRWKOVEES-UHFFFAOYSA-N 0.000 description 2
- FJKROLUGYXJWQN-UHFFFAOYSA-N 4-hydroxybenzoic acid Chemical compound OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 2
- 206010001742 Allergy to animal Diseases 0.000 description 2
- 206010002198 Anaphylactic reaction Diseases 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- 206010012438 Dermatitis atopic Diseases 0.000 description 2
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- NTYJJOPFIAHURM-UHFFFAOYSA-N Histamine Chemical compound NCCC1=CN=CN1 NTYJJOPFIAHURM-UHFFFAOYSA-N 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 108010063738 Interleukins Proteins 0.000 description 2
- 102000015696 Interleukins Human genes 0.000 description 2
- 241000222385 Phanerochaete Species 0.000 description 2
- 108010045362 Serum Globulins Proteins 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 210000001744 T-lymphocyte Anatomy 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000011149 active material Substances 0.000 description 2
- WNROFYMDJYEPJX-UHFFFAOYSA-K aluminium hydroxide Chemical compound [OH-].[OH-].[OH-].[Al+3] WNROFYMDJYEPJX-UHFFFAOYSA-K 0.000 description 2
- 208000003455 anaphylaxis Diseases 0.000 description 2
- 239000003146 anticoagulant agent Substances 0.000 description 2
- 229940127219 anticoagulant drug Drugs 0.000 description 2
- 201000008937 atopic dermatitis Diseases 0.000 description 2
- 235000019445 benzyl alcohol Nutrition 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- 238000000576 coating method Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 239000003623 enhancer Substances 0.000 description 2
- 238000011156 evaluation Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000007789 gas Substances 0.000 description 2
- 210000001035 gastrointestinal tract Anatomy 0.000 description 2
- 150000002617 leukotrienes Chemical class 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 239000008176 lyophilized powder Substances 0.000 description 2
- 238000004452 microanalysis Methods 0.000 description 2
- 239000000825 pharmaceutical preparation Substances 0.000 description 2
- 229940127557 pharmaceutical product Drugs 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 239000003223 protective agent Substances 0.000 description 2
- 239000011541 reaction mixture Substances 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 210000003491 skin Anatomy 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 229910001220 stainless steel Inorganic materials 0.000 description 2
- 239000010935 stainless steel Substances 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 230000009284 tracheal contraction Effects 0.000 description 2
- 210000005092 tracheal tissue Anatomy 0.000 description 2
- AXTGDCSMTYGJND-UHFFFAOYSA-N 1-dodecylazepan-2-one Chemical compound CCCCCCCCCCCCN1CCCCCC1=O AXTGDCSMTYGJND-UHFFFAOYSA-N 0.000 description 1
- 229940090248 4-hydroxybenzoic acid Drugs 0.000 description 1
- 241000238876 Acari Species 0.000 description 1
- WBZFUFAFFUEMEI-UHFFFAOYSA-M Acesulfame k Chemical compound [K+].CC1=CC(=O)[N-]S(=O)(=O)O1 WBZFUFAFFUEMEI-UHFFFAOYSA-M 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- 102100029348 CDGSH iron-sulfur domain-containing protein 2 Human genes 0.000 description 1
- 206010008479 Chest Pain Diseases 0.000 description 1
- 206010010356 Congenital anomaly Diseases 0.000 description 1
- 206010011224 Cough Diseases 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 208000000059 Dyspnea Diseases 0.000 description 1
- 206010013975 Dyspnoeas Diseases 0.000 description 1
- 241000588914 Enterobacter Species 0.000 description 1
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 description 1
- 108010053070 Glutathione Disulfide Proteins 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 239000007995 HEPES buffer Substances 0.000 description 1
- 239000004705 High-molecular-weight polyethylene Substances 0.000 description 1
- 101000989662 Homo sapiens CDGSH iron-sulfur domain-containing protein 2 Proteins 0.000 description 1
- 206010021033 Hypomenorrhoea Diseases 0.000 description 1
- 102100037850 Interferon gamma Human genes 0.000 description 1
- 108010074328 Interferon-gamma Proteins 0.000 description 1
- 102000004388 Interleukin-4 Human genes 0.000 description 1
- 108090000978 Interleukin-4 Proteins 0.000 description 1
- 108010002616 Interleukin-5 Proteins 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 1
- 239000004384 Neotame Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 241000700159 Rattus Species 0.000 description 1
- 208000037656 Respiratory Sounds Diseases 0.000 description 1
- 206010039085 Rhinitis allergic Diseases 0.000 description 1
- 239000004376 Sucralose Substances 0.000 description 1
- 210000004241 Th2 cell Anatomy 0.000 description 1
- 208000024780 Urticaria Diseases 0.000 description 1
- 206010047924 Wheezing Diseases 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000003187 abdominal effect Effects 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 239000000619 acesulfame-K Substances 0.000 description 1
- 230000002378 acidificating effect Effects 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 208000038016 acute inflammation Diseases 0.000 description 1
- 230000006022 acute inflammation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 208000030961 allergic reaction Diseases 0.000 description 1
- 201000010105 allergic rhinitis Diseases 0.000 description 1
- 230000001668 ameliorated effect Effects 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 0.000 description 1
- 210000003719 b-lymphocyte Anatomy 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
- 238000007906 compression Methods 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000428 dust Substances 0.000 description 1
- 230000001804 emulsifying effect Effects 0.000 description 1
- YQGOJNYOYNNSMM-UHFFFAOYSA-N eosin Chemical compound [Na+].OC(=O)C1=CC=CC=C1C1=C2C=C(Br)C(=O)C(Br)=C2OC2=C(Br)C(O)=C(Br)C=C21 YQGOJNYOYNNSMM-UHFFFAOYSA-N 0.000 description 1
- 238000013401 experimental design Methods 0.000 description 1
- 210000003608 fece Anatomy 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 230000007661 gastrointestinal function Effects 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- 235000013922 glutamic acid Nutrition 0.000 description 1
- 239000004220 glutamic acid Substances 0.000 description 1
- YPZRWBKMTBYPTK-BJDJZHNGSA-N glutathione disulfide Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@H](C(=O)NCC(O)=O)CSSC[C@@H](C(=O)NCC(O)=O)NC(=O)CC[C@H](N)C(O)=O YPZRWBKMTBYPTK-BJDJZHNGSA-N 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 229960001340 histamine Drugs 0.000 description 1
- 235000020256 human milk Nutrition 0.000 description 1
- 210000004251 human milk Anatomy 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 230000000968 intestinal effect Effects 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 229960003639 laurocapram Drugs 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 239000008297 liquid dosage form Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000005923 long-lasting effect Effects 0.000 description 1
- 230000004199 lung function Effects 0.000 description 1
- 238000013123 lung function test Methods 0.000 description 1
- 210000002540 macrophage Anatomy 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 235000019412 neotame Nutrition 0.000 description 1
- HLIAVLHNDJUHFG-HOTGVXAUSA-N neotame Chemical compound CC(C)(C)CCN[C@@H](CC(O)=O)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 HLIAVLHNDJUHFG-HOTGVXAUSA-N 0.000 description 1
- 108010070257 neotame Proteins 0.000 description 1
- 230000004768 organ dysfunction Effects 0.000 description 1
- 230000036284 oxygen consumption Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 235000011837 pasties Nutrition 0.000 description 1
- 210000003819 peripheral blood mononuclear cell Anatomy 0.000 description 1
- 230000004526 pharmaceutical effect Effects 0.000 description 1
- WVDDGKGOMKODPV-ZQBYOMGUSA-N phenyl(114C)methanol Chemical compound O[14CH2]C1=CC=CC=C1 WVDDGKGOMKODPV-ZQBYOMGUSA-N 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 210000003240 portal vein Anatomy 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011165 process development Methods 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000011514 reflex Effects 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 208000023504 respiratory system disease Diseases 0.000 description 1
- 238000001223 reverse osmosis Methods 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- 239000012898 sample dilution Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 210000002460 smooth muscle Anatomy 0.000 description 1
- 238000002791 soaking Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 239000001509 sodium citrate Substances 0.000 description 1
- NLJMYIDDQXHKNR-UHFFFAOYSA-K sodium citrate Chemical compound O.O.[Na+].[Na+].[Na+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O NLJMYIDDQXHKNR-UHFFFAOYSA-K 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 235000019408 sucralose Nutrition 0.000 description 1
- BAQAVOSOZGMPRM-QBMZZYIRSA-N sucralose Chemical compound O[C@@H]1[C@@H](O)[C@@H](Cl)[C@@H](CO)O[C@@H]1O[C@@]1(CCl)[C@@H](O)[C@H](O)[C@@H](CCl)O1 BAQAVOSOZGMPRM-QBMZZYIRSA-N 0.000 description 1
- 239000013595 supernatant sample Substances 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 125000003396 thiol group Chemical group [H]S* 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 230000029069 type 2 immune response Effects 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 235000015192 vegetable juice Nutrition 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P11/00—Drugs for disorders of the respiratory system
- A61P11/06—Antiasthmatics
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
- A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/52—Adding ingredients
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/70—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter
- A23L2/84—Clarifying or fining of non-alcoholic beverages; Removing unwanted matter using microorganisms or biological material, e.g. enzymes
-
- 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
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/26—Carbohydrates, e.g. sugar alcohols, amino sugars, nucleic acids, mono-, di- or oligo-saccharides; Derivatives thereof, e.g. polysorbates, sorbitan fatty acid esters or glycyrrhizin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/08—Antiallergic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/51—Bifidobacterium
- A23V2400/531—Lactis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K2035/11—Medicinal preparations comprising living procariotic cells
- A61K2035/115—Probiotics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Abstract
本发明提供一种乳双歧杆菌GKK2、含其的组合物及其改善过敏性气喘的用途。本发明的乳双歧杆菌具有耐酸、耐胆盐及/或耐热能力,其每毫升的菌落形成单位于pH3.2至pH2.0间的培养基中为约5x108cfu/ml、于添加0.3%胆盐的液态培养基中为约5x109cfu/ml及/或以特定温度加热时间大于0分钟至约15分钟间的液态培养基中为约5x107cfu/ml至5x109cfu/ml。本发明的包含该菌的组合物可用于降低呼吸道阻力、降低血清中非特异性抗体IgE含量、降低血清中特异性抗体IgE含量、减缓气管收缩程度及提升肝脏组织液中谷胱甘肽还原酶(GSH reductase)的含量,以改善过敏性气喘。
Description
技术领域
本发明关于一种用于改善过敏性气喘的乳酸菌、含其的组合物及其用途;更明确地说,是关于一种以乳双歧杆菌(Bifidobacterium lactis)制造包含该菌种的组合物,并将该组合物施予受施者,以用于降低呼吸道阻力、降低血清中非特异性抗体IgE含量、降低血清中特异性抗体IgE含量、减缓气管收缩程度及提升肝脏组织液中谷胱甘肽还原酶的含量,以改善过敏性气喘。
背景技术
过敏反应
过敏反应是指免疫系统对本来无害的物质产生过度反应,进而造成组织急性或慢性发炎,严重者甚至造成器官功能失调。常见过敏性疾病如过敏性气喘、过敏性鼻炎、荨麻疹、异位性皮肤炎…等。
过敏性气喘
过敏性气喘为一种呼吸道疾病,随着全球环境急遽恶化,气喘盛行率不断升高,主要症状为呼吸困难、咳嗽、胸闷、呼吸有喘鸣声,严重者可能致命。当环境中过敏原藉由各种方式进入人体后,首次会先经历致敏(Sensitization)的过程,意即当第一次接触过敏原(Allergen)的时候,过敏原会与特异性抗原细胞结合,当特异性抗原细胞将过敏原呈现给T淋巴球,T淋巴球分化成Th2细胞,进而释放出许多发炎相关细胞激素,且同时刺激B淋巴球制造抗体IgE(免疫球蛋白E),抗体IgE随着血液进入组织,连结于肥大细胞(mast cells)表面,此时即带有过敏体质。当人体再次暴露于相同过敏原时,过敏原会与肥大细胞表面的IgE结合,刺激肥大细胞释放出许多发炎相关介质,如组织胺、白三烯素(Leukotriene﹐LT)、白介素(Interleukin﹐IL)等,直接或间接造成呼吸道发炎反应。当发炎反应发生时,会使得呼吸道气管肿胀及气道周围平滑肌收缩,造成气管变窄,随着黏液腺体分泌增加,黏液一旦充满气管内部空隙,就会使气管急遽收缩,引发气喘。
谷胱甘肽
谷胱甘肽(Glutathione,GSH)是一种含γ-酰胺键和巯基的三肽,由谷氨酸、半胱氨酸及甘氨酸组成。此物质存在于几乎身体的每一个细胞,并具有维持身体正常的免疫系统的功能。谷胱甘肽还原酶能将谷胱甘肽氧化态(GSSG)还原成谷胱甘肽还原态(GSH),而得以增加体内GSH的浓度,达到强化免疫系统的作用。
双歧杆菌属
双歧杆菌属(Bifidobacterium)广泛存在于人和动物的消化道、阴道和口腔等环境中。其为革兰氏阳性、不运动、细胞呈杆状、一端有时呈分叉状且严格厌氧。此细菌早在1899年就从健康婴儿的粪便中分离出来,随后的研究发现,此细菌的部分特定菌株可作为益生菌以应用于食品、医药和饲料等领域。
先前研究有发表少数乳酸菌可促进INF-γ的表达,进而抑制过敏相关的IL-4、IL-5及特异性IgE的表达量,而能够调降Th2免疫反应,进而减缓过敏症状。亦有研究指出连续使用四个月的乳酸菌,可以使先天过敏体质的人的IFN-γ分泌增加而改善过敏。近年来临床试验结果甚至发现早期摄取益生菌,可减少婴儿异位性皮肤炎的产生或症状减轻,并相对减少过敏性气喘的发生机率。此外,先前学者以L. acidophilus进行双盲试验,发现能有效降低尘螨及花粉造成的过敏性气喘症状。然而,上述所述及的菌种皆无法改善过敏性气管的过度收缩的问题。
发明内容
本发明提供一种乳双歧杆菌(Bifidobacterium lactis),该菌具有耐酸、耐胆盐及/或耐热能力,其每毫升的菌落形成单位于pH3.2至pH2.0间的培养基中为约5x108cfu/ml、于添加0.3%胆盐的液态培养基中为约5x109cfu/ml及/或以特定温度加热时间大于0分钟至约15分钟间的液态培养基中为约5x107cfu/ml至5x109cfu/ml。
较佳地,该菌株为乳双歧杆菌(Bifidobacterium lactis) GKK2,且以保藏编号CGMCC No. 15205保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)(地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所),保藏日期:2018年01月12日,分类命名:乳双歧杆菌(Bifidobacterium lactis)(保藏编号BCRC910826保藏于财团法人食品工业发展研究所)。
本发明为有效改善过敏性气喘提供一种组合物,该组合物包含乳双歧杆菌(Bifidobacterium lactis) GKK2,该菌株以保藏编号CGMCC No. 15205保藏于中国微生物菌种保藏管理委员会普通微生物中心(保藏编号BCRC910826保藏于财团法人食品工业发展研究所)。
本发明另提供一种改善过敏性气喘的组合物,其包含乳双歧杆菌(Bifidobacterium lactis) GKK2的活性物质,其中乳双歧杆菌GKK2以保藏编号CGMCC No.15205保藏于中国微生物菌种保藏管理委员会普通微生物中心(保藏编号BCRC910826保藏于财团法人食品工业发展研究所)。
较佳地,所述活性物质是以下列方法制备:
(a)取乳酸菌菌落(colony)接种于培养基以进行固态培养,形成乳酸菌菌体;及
(b)将步骤(a)培养的乳酸菌菌体接种于液体培养基以进行液态培养。
较佳地,所述方法进一步包含下列步骤:
(c)将步骤(b)培养的含乳酸菌菌体的液态培养基离心以获得菌泥;及
(d)将步骤(c)所得的菌泥进行冷冻干燥。
较佳地,步骤(b)的温度为35至50℃、通气量为0至1vvm氮气或二氧化碳、转速为10至100rpm及/或培养天数16至24小时。
较佳地,步骤(d)的冷冻干燥的预冻温度为-196至-40℃。
较佳地,上述组合物包含一种选自下列群组的添加剂:赋型剂、防腐剂、稀释剂、填充剂、吸收促进剂、甜味剂或其组合。
较佳地,一种组合物,其包含乳双歧杆菌GKK2或GKK2的活性物质,上述组合物为药品、饲料、饮料、营养补充品、乳制品、食品或保健食品。
较佳地,上述组合物的形态为粉剂、锭剂、造粒、栓剂、微胶囊、安瓶、液剂喷剂或塞剂。
本发明经过实验证实乳双歧杆菌(Bifidobacterium lactis)的菌株 GKK2具有可用于改善过敏性气喘的用途。
较佳地,上述用途的改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的呼吸道阻力下降。
较佳地,上述用途的改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的血清中非特异性抗体IgE含量下降。
较佳地,上述用途的改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的血清中特异性抗体IgE含量下降。
较佳地,上述用途的改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的气管收缩程度变小。
较佳地,上述用途的改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的肝脏组织液中谷胱甘肽还原酶(GSH reductase)的含量提升。
附图说明
图1显示耐酸试验结果的菌落数目。
图2显示耐胆盐试验的菌落数目。
图3显示耐热试验结果的菌落数目。
图4显示实验时程。
图5显示呼吸道阻力测试结果。
图6显示血清中非特异性抗体IgE含量。
图7显示血清中特异性抗体IgE含量。
图8显示气管组织切片。
图9显示谷胱甘肽还原酶(GSH reductase)的含量。
用于专利程序的微生物保存:
本发明的乳双歧杆菌GKK2:
保藏日期:2018年01月12日;
保藏单位:中国微生物菌种保藏管理委员会普通微生物中心(CGMCC);
保藏单位地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所;
保藏编号:CGMCC No. 15205;
分类命名:乳双歧杆菌(Bifidobacterium lactis)。
具体实施方式
菌种来源
本实验中所使用的乳酸菌是双歧杆菌属的乳双歧杆菌(Bifidobacterium lactis),又称为乳双歧杆菌。在一较佳的实施态样中,此菌种分离来自哺育母乳的三个月大健康男婴的肠道检体,且该男婴从出生至采样其间皆无使用疾病用药与或益生菌相关产品等可能影响肠胃道功能及检体菌相的药品或保健产品。该检体外观正常且呈黄褐色糊状,符合母乳便的特征。检体产生后,随即将其放入含有厌氧产气包的厌氧罐相关设备中,以维持检体中厌氧菌的活性,并于冷藏环境下运送至实验室进行菌种分离筛选作业。在一较佳的实施态样中,此菌种在2018年01月12日以保藏编号CGMCC No. 15205保藏于中国微生物菌种保藏管理委员会普通微生物中心(CGMCC)(地址:北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所),保藏日期:2018年01月12日,分类命名:乳双歧杆菌(Bifidobacterium lactis)(2018年1月12日以保藏编号BCRC910826保藏于财团法人食品工业发展研究所)。
筛选培养
含检体的厌氧缸在厌氧环境的厌氧操作台中开封,以灭菌的药匙取样约2克,将其放入无菌生理食盐水200毫升中均质。均质后的检体稀释至104~107倍率,接着取出检体稀释液以TOS-MUP 培养基进行倾注培养。利用TOS-MUP 培养基针对检体中比菲德氏菌进行筛选分离,有效地排除检体中Enterobacter干扰,以改善传统BIM-25培养基的缺点,比菲德氏菌筛选率达100%。
经倾注培养法培养后的平板置入厌氧缸中,打开氧气消耗用的厌氧气体包密封厌氧缸,接着移出厌氧操作台,放置于37±0.5℃恒温培养箱中培养44~48小时。
培养后厌氧缸移至厌氧操作台中进行开封与单一菌株的挑选及保存,每个检体个别挑选出约100株以上单株菌落进行分离培以保存与进行体外试验细胞评估,以外周血单个核细胞、脾脏初代细胞、肥大细胞体外评估与CISD2长寿基因活化试验,经多重体外试验进行筛选测试,筛选出具有提升免疫力、舒缓体内过敏反应及皮肤过敏反应的高机能性乳酸菌株。以分离得到一株具有多重生理功效改善的新颖性益生菌菌株,可用于发酵奶类或蔬果汁及制造益生菌粉直接服用,以改善许多生理上反应的异常现象。
表现型分析
以耐酸、耐胆盐与耐热试验比较GKK2的乳酸菌和其他菌种间表现型差异的研究。
耐酸试验
将GKK2、从食品工业发展研究所生物资源保存与研究中心购入的BCRC 17394、动物双歧杆菌乳酸亚种Bi 04(B. animalis subsp. LactisBi 04,以编号ATCC SD 5219保藏于美国典型菌种保存中心)、动物双歧杆菌乳亚种BB-12(B.animalis subsp. LactisBB-12,以编号DSM 15954保藏于德国微生物菌种保存中心)与动物双歧杆菌乳亚种Bi07(B.animalis subsp. LactisBi07,以编号ATCC SD 5220保藏于美国典型菌种保存中心)共5种菌株活化。藉由添加HCl 至MRS液态培养基中,将酸碱值约为pH 6.5的原始MRS液态培养基,调整该培养基的酸碱度为另3种不同酸碱度:约pH 3.2、pH 2.4与pH 2.0。将菌株接种在这些不同酸碱值的培养基,于37℃下培养3小时后,计数菌落形成数目。
结果如图1所示,于原始pH培养下(约pH 6.5),GKK2与其他4种菌株的菌数皆可达到5x109cfu/ml。于酸碱值为pH 3.2时,全部菌株的菌数略为下降,GKK2与其他4种菌株相较未出现显着差异。当酸碱值下降至pH 2.4跟pH 2.0时,BCRC 17394、BB-12与Bi07菌数降至约6~7次方,都显着低于菌数维持于8次方以上的GKK2(P<0.05)。据此,在酸性环境下GKK2的活菌数显着多于他种菌株,此实验结果说明GKK2的耐酸能力相较于其他菌株更佳,故GKK2通过胃部时抵御胃酸的能力更佳。
耐胆盐试验
将GKK2、从食品工业发展研究所生物资源保存与研究中心购入的BCRC 17394、动物双歧杆菌乳酸亚种Bi 04(B. animalis subsp. LactisBi 04,以编号ATCC SD 5219保藏于美国典型菌种保存中心)、动物双歧杆菌乳亚种BB-12(B.animalis subsp. LactisBB-12,以编号DSM 15954保藏于德国微生物菌种保存中心)与动物双歧杆菌乳亚种Bi07(B.animalis subsp. LactisBi07,以编号ATCC SD 5220保藏于美国典型菌种保存中心)共5种菌株活化。将这些菌种接种于含0.3%胆盐的MRS液态培养基中,于37℃下浸泡半小时后,观察并计数菌数。
结果如图2所示,于原始MRS 液态培养基培养下,GKK2与其他4种菌株的菌数皆可达到5x109cfu/ml。在添加有0.3%胆盐的MRS中,BCRC 17394、Bi 04与Bi07的菌数都显着低于GKK2菌数(P<0.05),BB-12的菌数则与GKK2统计上无显着差异。据此,在胆盐环境下GKK2的活菌数显着多于他种菌株,此实验结果说明GKK2的耐胆盐能力相较于其他菌株更佳,故GKK2通过体内消化道时抵御胆盐的能力更佳。
耐热试验
将GKK2、从食品工业发展研究所生物资源保存与研究中心购入的BCRC 17394、动物双歧杆菌乳酸亚种Bi 04(B. animalis subsp. LactisBi 04,以编号ATCC SD 5219保藏于美国典型菌种保存中心)、动物双歧杆菌乳亚种BB-12(B.animalis subsp. LactisBB-12,以编号DSM 15954保藏于德国微生物菌种保存中心)与动物双歧杆菌乳亚种Bi07(B.animalis subsp. LactisBi07,以编号ATCC SD 5220保藏于美国典型菌种保存中心)共5种菌株活化。将这些菌种接种于 MRS液态培养基中,分别于70℃下加热5、10、15分钟后,观察并计数菌数。
结果如图3所示,于未加热情况下(0分钟),GKK2与其他4种菌株的菌数皆可达到5x109cfu/ml。在70℃下加热5分钟后,BCRC 17394、Bi 04与Bi07的菌数都显着低于GKK2菌数(P<0.05),BB-12的菌数则与GKK2统计上无显着差异。而在70℃下加热15分钟后,其他4株菌的菌数都显着低于GKK2菌数(P<0.05)。据此,在高温环境下GKK2的活菌数显着多于他种菌株,此实验结果说明GKK2的耐热能力较好。况且,乳酸菌普遍都不耐热而须保存于低温下维持活性,此试验甚至显示GKK2具有耐高温并于常温下久放的特性,而对于将GKK2应用于工艺开发非常有帮助。
菌种培养
将上述的乳双歧杆菌,勾取其菌落(colony)接种于固态培养基上以活化菌种。在一较佳的实施态样中,所述固态培养基为MRS琼脂。待菌体生长完成后,将新鲜的菌体连同固态培养基接入于含有液态培养基的锥形瓶中进行液态培养。在一较佳的实施态样中,于温度35至50℃下、通气量0至1vvm氮气或二氧化碳、速率10至100rpm的条件下液态培养。在一较佳的实施态样中,液态培养的时间为16至24小时,更佳为18小时。在一较佳的实施态样中,液态培养基为MRS液态培养基。在一较佳的实施态样中,液态培养基的配方如下表1所示。
表1
冻干粉制备
待乳酸菌的菌种于液态培养完成生长后,收集包含有乳酸菌菌体的液态培养基进行离心以获得菌泥。在一较佳的实施态样中,包含有乳酸菌菌体的液态培养基以速率1000至15000rpm进行离心。将取得的菌泥与保护剂(保护剂为6-30%的脱脂奶粉)混合后冷冻干燥,冻干后置于低温保存。在一较佳的实施态样中,冷冻干燥的预冻温度设定于-196至-40℃。在一较佳的实施态样中,冷冻干燥时间为16至72小时。在一较佳的实施态样中,保存温度为-30℃至0℃。保存的乳酸菌冻干粉作为用于以下细胞实验的原料,意即为本案所请求保护的乳酸菌的活性物质的其中一态样。本案所请求保护的乳酸菌活性物质的态样,亦包含前述将菌体进行液态培养后所得的含菌体的培养液状态。
实验动物与设计
本实验动物选购Balb/c品种的5周大雄鼠共24只。小鼠饲养于小鼠专用房内的独立IVC (individual ventilated caging system)小鼠饲育系统,动物房的室温维持于22± 2℃、相对湿度保持在湿度40~60%,光照周期以自动定时器控制07:00至19:00为黑暗期(dark period),19:00至07:00为光照期(light period)。饲料及无菌逆渗透水均任由小鼠自由取食。小鼠饲养2周适应环境后,才开始进行实验。
动物实验中可通过致敏(sensitization)来建立动物过敏模式。本实验以过敏原作为抗原,搭配氢氧化铝(Aluminum hydroxide)作为佐剂,藉由皮下注射将过敏原注入以致敏动物。为确立动物过敏模式的建立,在注射抗原一周前及注射抗原一周后采血,从血液样本中取得血清以分析抗原特异性抗体浓度。必要时须致敏三至四次,使血清中的抗原特异性抗体浓度达到过敏模式建立的标准,始能进行后续实验检测。呼吸道过敏的动物模式则采用喷雾致敏法来建立,使动物吸入过敏原而从呼吸道来接触过敏原。
卵白蛋白抗原诱导小鼠过敏及气喘,是通过将卵白蛋白抗原(OVA, 500 μg/ml)与不完全佐剂(Incomplete Freund’s adjuvant)以1:1等量均匀混合乳化后,以无菌过滤膜进行过滤后立即使用。各组小鼠于实验第30、40及50日,皆进行此过敏原的皮下注射,并且于第45天及第55天以喷雾2% OVA过敏原方式,进行10分钟呼吸道吸入(inhalation)以诱导小鼠产生过敏气喘。
本实验的24只5周龄Balb/c雄鼠,分为喂食生理食盐水的对照组、以卵白蛋白(OVA)诱导致敏的致敏组,以及先以卵白蛋白(OVA)诱导致敏再喂食乳双歧杆菌GKK2的高、低剂量组的实验组,共4组,每组6只。乳双歧杆菌GKK2的喂食采小鼠管灌,灌食剂量的计算依据实验动物与人体表面积比的等效剂量换算系数,其中小鼠与人的换算系数为12.3。小鼠每周秤量体重一次,每组计算出平均体重后,依厂商提供的样品成人(以体重60千克计算)每日服用剂量为每20 mg 与100 mg(约为每人每日100与300亿个活菌数),加以换算每天管灌小鼠的剂量,公式如下所示:
小鼠每日喂食产品剂量(g)={成人剂量/60千克(成人体重)}×小鼠体重(千克)×12.3(小鼠与人的换算系数)
经上述公式计算出实验组喂食乳双歧杆菌GKK2的剂量如下表2所示:
表2
以0.9 %生理食盐水加入上述剂量的乳双歧杆菌GKK2配制喂食小鼠的实验样品。不锈钢喂食针平时浸泡在75 %酒精溶液中,使用前再以0.9 %生理食盐水清洗润湿。各组小鼠分别以1 ml无菌塑胶针筒套上不锈钢喂食针,分别管灌喂食小鼠0.2 ml体积的生理食盐水(对照组)或不同浓度的乳双歧杆菌GKK2溶液。各组小鼠每天管灌喂食上述样品,执行期间共连续投予60天。详细的实验时程如图4。
于第62天牺牲小鼠以取得血液并分离与保存血清。收集小鼠腹腔巨噬细胞后,剪开腹部皮肤,以1 ml注射器于肝门静脉收集血液样本。收集小鼠全血检体,置于含有抗凝血剂的试管中,待进行血液中白血球分类计数。另外,收集小鼠全血检体于无含抗凝血剂的试管中,室温静置1小时后以1000 rpm的速度离心10分钟,取得上清液检体即为血清。血清长期保存于-80℃,待日后分析血清中免疫球蛋及过敏原诱发的特异性抗体。
呼吸道阻力测试
小鼠于牺牲前一天,以Buxco公司出产的全体肺功能检测系统(Whole-bodyplethysmograghy,WBP),进行非侵入性的小鼠呼吸道阻力变化的测量。阻力变化是依据小鼠呼吸过程中测得的数值经电脑分析后,得到一个作为肺功能指标的增强性间歇值(enhanced pause,Penh),若数值越高则代表呼吸道阻力越大。首先,将小鼠放进动物舱中,测量小鼠的基础Penh值三分钟。接着,将小鼠暴露在雾化的0.9% NaCl三分钟后,测量Penh值三分钟。再来,将小鼠暴露在雾化的非特异性气管收缩刺激剂乙酰甲基胆素(methacholine)(6.25 mg/ml)三分钟后,测量Penh值(PenhMch)三分钟,并以逐渐增高浓度的乙酰甲基胆素(12.5 mg/ml、25 mg/ml、50 mg/ml)重复上述步骤。实验结果于本实验中是以Penh Ratio来表示,数据分析是将各浓度乙酰甲基胆素所得到的PenhMch读值除以给予生理食盐水的PenhNaCl读值来计算。
血清中免疫球蛋白IgE含量分析
利用三明治法酶联免疫吸附分析(sandwich-ELISA),定量血清中免疫球蛋白IgE抗体含量。首先以涂覆液(coating buffer)(pH 9.6)配制各种含适量的anti-mice单株抗体,覆盖于96孔微量分析盘(Nunc-Immuno plate)上,静置1小时后,以清洗液(Washbuffer)洗去未结合的单株抗体,再以200μl/well的阻断液(blocking buffer)填补未被单株抗体结合的部位。室温下反应30分钟后,再以清洗液(wash buffer)冲洗,然后加入100 μl小鼠血清或已知浓度免疫球蛋白IgE标准品。小鼠血清或是免疫球蛋白IgE标准品经过室温1小时反应后,以清洗液(wash buffer)冲洗,再加入适当浓度连结HRP的抗免疫球蛋白IgE的二级抗体100 μl/well。于室温反应30分钟后,以清洗液(wash buffer)冲洗,接着以100 μl TMB受质反应,经15分钟呈色后,加入100μl 2N H2SO4终止呈色反应,于450nm波长测定其吸光值。根据免疫球蛋白IgE标准品与吸光值所显示的标准曲线,经过回归运算后得到的公式,可用内插法来推算血清中抗体的浓度。
血清中过敏原诱发的特异性抗体IgE含量分析
利用三明治法酶联免疫吸附分析(sandwich-ELISA),定量血清中过敏原诱发的特异性IgE含量。首先以涂覆液(coating buffer)(pH 9.6)配制含适量的anti-mice单株抗体并添加100μg/mL卵白蛋白(OVA)抗原,覆盖于96孔微量分析盘(Nunc-Immuno plate)上,静置1小时后,以清洗液(wash buffer)冲洗未结合的单株抗体,再以200 μl/well的阻断液(blocking buffer)填补未被单株抗体结合的部位。室温下反应30分钟后,再以清洗液(wash buffer)冲洗,然后加入100 μl小鼠血清或已知浓度的免疫球蛋白IgE标准品。小鼠血清或是免疫球蛋白IgE标准品经过室温1小时反应后,以清洗液(wash buffer)冲洗,再加入适当浓度连结HRP的抗免疫球蛋白IgE的二级抗体100 μl/well。于室温反应30分钟后,以清洗液(wash buffer)冲洗,接着以100 μl TMB受质反应,经15分钟呈色后,加入100 μl 2NH2SO4终止呈色反应,于450nm波长测定其吸光值。根据免疫球蛋白IgE标准品与吸光值所显示的标准曲线,经过回归运算后得到的公式,可用内插法来推算血清中过敏原诱发的特异性抗体的浓度。
组织切片
将气管组织以石蜡包埋和组织切片后,进行苏木素与伊红染色(hematoxylin andeosin stain,H&E stain)。于显微镜下进行型态观察。
谷胱甘肽还原酶(GSH reductase)的含量分析
将小鼠的肝脏定量放入HEPES 缓冲液细胞培养液中,实验前样品需于-80℃保存,等待谷胱甘肽还原酶(GSH reductase)的分析,以市售的ELISA方法分析其含量。
生物统计分析
本份测试报告的实验结果以mean±SD表示,并以SPSS 12.0套装软件于电脑中进行统计分析,以邓肯氏多变域测验(Duncan’s multiple range test)测定各组间的差异,显着性差异的判断标准为p<0.05,实验结果以SigmaPlot 10.0软件作图。
实验结果
针对呼吸道阻力测试,各组的实验结果显示如表3并统计如图5:
表3
从上述结果来看,当雾化气管收缩刺激剂乙酰甲基胆素浓度增加时,以OVA腹腔注射及吸入的致敏组的呼吸道阻力大幅增加,且明显高于对照组,说明致敏组以OVA致敏的小鼠模式成功建立。致敏小鼠给予GKK2的实验组,当雾化气管收缩刺激剂乙酰甲基胆素浓度增加时,呼吸道阻力上升幅度较低。说明显示给予GKK2后可减缓因OVA过敏原所引发的呼吸道压力,意即可改善气喘的压迫程度。此外,当提高给予的GKK2浓度时,呼吸道阻力上升幅度更小,说明GKK2减缓故敏性气喘的功效有浓度依存性。
针对血清中非特异性抗体IgE含量,实验结果显示如表4并统计如图6:
表4
致敏小鼠(致敏组)血清中非特异性抗体IgE抗体与对照组相比显着增加。管灌喂食投予GKK2的致敏小鼠(高、低剂量的实验组),体内血清中的非特异性抗体IgE含量,相对于致敏组显着降低(p<0.05),而接近对照组的含量数值。说明给予GKK2能降低血清中非特异性抗体IgE抗体浓度,而得以改善过敏症状。
针对血清中特异性抗体IgE含量,实验结果显示如表5并统计如图7:
表5
致敏小鼠(致敏组)的特异性抗体IgE与对照组相比明显提高。管灌喂食GKK2的致敏小鼠(高、低剂量的实验组),体内血清中的特异性IgE抗体含量,相对于致敏组显着降低(p<0.05)。说明给予GKK2能降低血清中特异性抗体IgE抗体浓度,而得以改善过敏症状。
针对组织切片,实验结果显示如图8。致敏的小鼠(致敏组)与对照组相比,气管明显缩小。管灌喂食GKK2的致敏小鼠(高、低剂量的实验组),气管收缩相较于致敏组来的不明显,且与对照组相比无明显差异。说明给予GKK2能改善气管收缩的症状,而具有降低气喘发作率的功效。
针对谷胱甘肽还原酶(GSH reductase)的含量,实验结果显示如表6并统计如图9:
表6
受致敏后的小鼠组别(气喘组),其谷胱甘肽还原酶含量大幅降低,管喂较高剂量GKK2后再致敏的小鼠组别,谷胱甘肽还原酶含量大幅提高,显着高于气喘组(p<0.05)。说明给予GKK2能提升免疫功能以改善过敏体质,而具有降低气喘发作的功效。
综上所述,经由前述各项实验结果可证,GKK2能够改善过敏性气喘所造成的气管收缩致使管腔缩小的问题,同时降低血液中因过敏增加的非特异性及特异性IgE抗体的含量,还能提高体内谷胱甘肽还原酶含量。说明乳双歧杆菌GKK2具有改善过敏性气喘的功效。
本发明提供一种组合物,包含有GKK2且发挥减缓过敏性气喘的功效。
所述组合物进一步包含添加剂。在一较佳的实施态样中,添加剂可为赋型剂、防腐剂、稀释剂、填充剂、吸收促进剂、甜味剂、或其组合。赋型剂可选自柠檬酸钠、碳酸钙、磷酸钙、蔗糖或其组合。防腐剂可延长医药组合物的储藏期限,例如苯甲醇、对羟基苯甲酸(parabens)。稀释剂可选自水、乙醇、丙二醇、甘油或其组合。填充剂可选自乳糖、牛乳糖、高分子量聚乙二醇或其组合。吸收促进剂可选自二甲基亚砜(DMSO)、月桂氮卓酮、丙二醇、甘油、聚乙二醇或其组合。甜味剂可选自安塞蜜(Acesulfame K)、阿斯巴甜(aspartame)、糖精(saccharin)、三氯蔗糖/蔗糖素(sucralose)、纽甜(neotame)或其组合。除上述所列举的添加剂以外,在不影响乳酸菌的活性物质的医药效果前提下,可依需求选用适合的其他添加剂。
所述组合物于医药领域中可开发为不同商品。
在一较佳实施态样中,提供一种组合物,其含有乳双歧杆菌GKK2,其为药品、饲料、饮料、营养补充品、乳制品、食品或保健食品。
所述组合物可根据受施予者的需要,而采用不同形态。在一较佳实施态样中,所述组合物的形态为粉剂、锭剂、造粒、栓剂、微胶囊、安瓶(ampoule/ampule)、液剂喷剂或塞剂。
本发明的组合物可使用于动物或是人类。在不影响乳酸菌的活性物质发挥效果的前提下,包含乳酸菌的活性物质的组合物可制为任何药物型态,并根据药物型态以适用的途径施予动物或人类。
组合物制备
本发明的GKK2若应用于食品用途,则以下组合物1的态样作为例示性实例。
组合物1:取GKK2的冻干粉作为乳酸菌的活性物质(20 wt%),与作为防腐剂的苯甲醇(8wt%)、作为稀释剂的甘油(7 wt%)充分混合,并溶于纯水(65 wt%)中,存放于4℃备用。前述wt%是指各成分占组合物总重的比例。
本发明的GKK2若以液体剂型应用于医药用途,则以下组合物2的态样作为例示性实例。
组合物2:取GKK2的冻干粉作为乳酸菌的活性物质(20 wt%),与作为防腐剂的苯甲醇(8wt%)、作为稀释剂的甘油(7 wt%)、作为稀释剂的蔗糖(10 wt%)充分混合,并溶于纯水(55 wt%)中,存放于4℃备用。前述wt%是指各成分占组合物总重的比例。
Claims (18)
1.一种乳双歧杆菌(Bifidobacterium lactis),该菌为乳双歧杆菌GKK2,其保藏编号为CGMCC No. 15205,保藏于中国微生物菌种保藏管理委员会普通微生物中心。
2.一种组合物,其包含权利要求1所述的乳双歧杆菌GKK2。
3.一种组合物,其包含权利要求1所述的乳双歧杆菌GKK2的活性物质;
其中所述活性物质是以下列方法制备得到的:
(a)取乳双歧杆菌GKK2的菌落接种于培养基以进行固态培养,形成乳双歧杆菌菌体;
(b)将步骤(a)培养的乳双歧杆菌菌体接种于液体培养基以进行液态培养;
(c)将步骤(b)培养的含乳双歧杆菌菌体的液态培养基离心以获得菌泥;及
(d)将步骤(c)所得的菌泥进行冷冻干燥。
4.如权利要求3所述的组合物,其中步骤(b)的培养条件为:温度为35至50℃,通气量为0至1vvm氮气或二氧化碳,转速为10至100rpm及培养时间16至24小时。
5.如权利要求3所述的组合物,其中步骤(d)的冷冻干燥的预冻温度为-196至-40℃。
6.如权利要求2至5中任一项所述的组合物,其包含一种选自下列群组的添加剂:赋型剂、防腐剂、稀释剂、填充剂、吸收促进剂、甜味剂或其组合。
7.如权利要求2至5中任一项所述的组合物,其为药品、饲料或食品。
8.如权利要求2至5中任一项所述的组合物,其为饮料。
9.如权利要求2至5中任一项所述的组合物,其为营养补充品。
10.如权利要求2至5中任一项所述的组合物,其为乳制品。
11.如权利要求2至5中任一项所述的组合物,其为保健食品。
12.如权利要求2至5中任一项所述的组合物,其形态为粉剂、锭剂、造粒、栓剂、微胶囊、安瓶、液剂喷剂或塞剂。
13.保藏编号为CGMCC No. 15205的乳双歧杆菌GKK2在制备用于改善过敏性气喘的医药组合物中的用途。
14.如权利要求13所述的用途,其中所述改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的呼吸道阻力下降。
15.如权利要求13所述的用途,其中所述改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的血清中非特异性抗体IgE含量下降。
16.如权利要求13所述的用途,其中所述改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的血清中特异性抗体IgE含量下降。
17.如权利要求13所述的用途,其中所述改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的气管收缩程度变小。
18.如权利要求13所述的用途,其中所述改善过敏性气喘是相对于未施予乳双歧杆菌GKK2的受施者,施予乳双歧杆菌GKK2的受施者的肝脏组织液中谷胱甘肽还原酶的含量提升。
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
TW107128653 | 2018-08-16 | ||
TW107128653A TWI709408B (zh) | 2018-08-16 | 2018-08-16 | 乳雙歧桿菌gkk2、含其之組合物及改善過敏性氣喘之用途 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN110835614A CN110835614A (zh) | 2020-02-25 |
CN110835614B true CN110835614B (zh) | 2023-08-25 |
Family
ID=69524283
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201811083035.0A Active CN110835614B (zh) | 2018-08-16 | 2018-09-17 | 乳双歧杆菌gkk2、含其的组合物及其改善过敏性气喘的用途 |
Country Status (6)
Country | Link |
---|---|
US (1) | US11590180B2 (zh) |
JP (1) | JP6985342B2 (zh) |
CN (1) | CN110835614B (zh) |
MY (1) | MY192498A (zh) |
SG (1) | SG10201907549UA (zh) |
TW (1) | TWI709408B (zh) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111560335B (zh) * | 2020-05-26 | 2022-09-23 | 北京科拓恒通生物技术股份有限公司 | 一株用于缓解哮喘的乳双歧杆菌及其应用 |
CN115505543B (zh) * | 2022-09-28 | 2024-01-26 | 天津小薇生物科技有限公司 | 一株乳双歧杆菌br001及其发酵方法与改善过敏的应用 |
CN116083323A (zh) * | 2023-03-16 | 2023-05-09 | 威凯海思(山东)生物工程有限公司 | 一株可缓解过敏反应的乳双歧杆菌hc2786及其产品和应用 |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103898018A (zh) * | 2014-03-28 | 2014-07-02 | 北京和美科盛生物技术有限公司 | 一种双歧乳杆菌的高密度培养方法及其冻干菌粉制备 |
CN104531590A (zh) * | 2015-01-14 | 2015-04-22 | 贵州大学 | 一株香猪源性降胆固醇、耐氧双歧杆菌bz11 |
CN107050063A (zh) * | 2016-11-08 | 2017-08-18 | 江西益盟科技有限公司 | 治疗过敏体质的乳酸菌组合物及其制备方法 |
Family Cites Families (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2139502B1 (en) * | 2007-03-26 | 2014-06-18 | DuPont Nutrition Biosciences ApS | Lactobacillus and bifidobacterium against birch allergy |
CN105765058A (zh) * | 2013-11-25 | 2016-07-13 | 雀巢产品技术援助有限公司 | 经热处理的乳酸双歧杆菌ncc2818制剂减轻变态反应的表现 |
RU2755083C2 (ru) * | 2015-01-27 | 2021-09-15 | ДюПон НЬЮТРИШН БАЙОСАЙЕНСИЗ АпС | Иммуномодулирующая композиция, содержащая бифидобактерии |
-
2018
- 2018-08-16 TW TW107128653A patent/TWI709408B/zh active
- 2018-09-17 CN CN201811083035.0A patent/CN110835614B/zh active Active
-
2019
- 2019-08-14 JP JP2019148830A patent/JP6985342B2/ja active Active
- 2019-08-15 US US16/541,964 patent/US11590180B2/en active Active
- 2019-08-16 MY MYPI2019004723A patent/MY192498A/en unknown
- 2019-08-16 SG SG10201907549UA patent/SG10201907549UA/en unknown
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN103898018A (zh) * | 2014-03-28 | 2014-07-02 | 北京和美科盛生物技术有限公司 | 一种双歧乳杆菌的高密度培养方法及其冻干菌粉制备 |
CN104531590A (zh) * | 2015-01-14 | 2015-04-22 | 贵州大学 | 一株香猪源性降胆固醇、耐氧双歧杆菌bz11 |
CN107050063A (zh) * | 2016-11-08 | 2017-08-18 | 江西益盟科技有限公司 | 治疗过敏体质的乳酸菌组合物及其制备方法 |
Non-Patent Citations (1)
Title |
---|
肠道菌群制剂对新生儿哮喘57例防治效果观察;曾双志;《现代诊断与治疗》;20151231;第26卷(第13期);第2956-2957页 * |
Also Published As
Publication number | Publication date |
---|---|
MY192498A (en) | 2022-08-24 |
JP2020074751A (ja) | 2020-05-21 |
SG10201907549UA (en) | 2020-03-30 |
JP6985342B2 (ja) | 2021-12-22 |
TWI709408B (zh) | 2020-11-11 |
US11590180B2 (en) | 2023-02-28 |
CN110835614A (zh) | 2020-02-25 |
US20200054691A1 (en) | 2020-02-20 |
TW202008994A (zh) | 2020-03-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN110835614B (zh) | 乳双歧杆菌gkk2、含其的组合物及其改善过敏性气喘的用途 | |
CN107058158A (zh) | 一种提高犬免疫力降低腹泻发病率的复合益生菌剂及其制备和应用 | |
CN107164263A (zh) | 一种可调节肠道功能、预防结肠炎的发酵乳杆菌hy01及其用途 | |
CN110643524B (zh) | 一种具有胃肠道粘膜保护作用的复合益生菌制剂及其应用 | |
CN113005067B (zh) | 多功能复合益生菌制剂及其制备方法 | |
CN113943681B (zh) | 一株降低炎症反应且具有缓解便秘作用的长双歧杆菌 | |
CN114848685A (zh) | 抗过敏益生菌组合物及其应用、以及益生菌中药发酵物 | |
CN111329884B (zh) | 植物乳杆菌bc299在炎症性肠病、精神性问题的药物、食品中的应用 | |
CN114774315B (zh) | 鼠李糖乳杆菌菌株LRa05在制备增强免疫力制品和/或缓解湿疹制品方面的用途 | |
CN113881597B (zh) | 一株能够提高吲哚丙烯酸以调节特异性IgE的罗伊氏乳杆菌 | |
CN111685255B (zh) | 一种增强免疫功能的益生菌固体饮料及其制备方法 | |
US11058733B2 (en) | Active substances of Bifidobacterium lactis GKK2, composition comprising the same and method of promoting longevity using the same | |
CN114468306B (zh) | 凝结芽孢杆菌bc99在制备缓解结肠炎制品或免疫调节制品方面的用途 | |
CN116445356A (zh) | 一种调节肠道菌群及增强免疫力的动物双歧杆菌乳亚种ba67及其应用 | |
CN113005066B (zh) | 抗过敏、增加免疫力、降血糖、降脂减肥的复合双歧杆菌制剂及其制备方法 | |
CN116024129A (zh) | 一株能够与幽门螺杆菌共聚集的卷曲乳杆菌及其应用 | |
CN114921383A (zh) | 一种具有清除胆固醇功能的益生菌制剂及其制备方法 | |
CN112029676B (zh) | 一种有利于提高免疫力的益生菌组合及其应用 | |
CN116240152B (zh) | 一种鼠李糖乳杆菌sf-l30及其在制备清肺消炎产品中的应用 | |
CN113913330B (zh) | 一株调节OVA特异性IgE的植物乳杆菌及其应用 | |
RU2176668C1 (ru) | ШТАММ БАКТЕРИЙ LACTOBACILLUS ACIDOPHILUS N.V.EP 317/402 "НАРИНЭ" ТНСи, ИСПОЛЬЗУЕМЫЙ ПРИ ПРИГОТОВЛЕНИИ ЛЕЧЕБНО-ПРОФИЛАКТИЧЕСКИХ ПРЕПАРАТОВ ДЛЯ НОРМАЛИЗАЦИИ КИШЕЧНОЙ МИКРОФЛОРЫ | |
CN114317310B (zh) | 一种抗过敏的婴儿双歧杆菌制剂及其制备方法 | |
CN116656526B (zh) | 一株植物乳植杆菌jf4及其在制备降血糖和降胆固醇食品药品中的应用 | |
KR102244008B1 (ko) | 글루텐 분해능을 가지는 락토바실러스 파라카제이 glu70 균주의 사균체를 유효성분으로 함유하는 글리아딘으로 야기된 염증성 장 질환의 예방, 개선 또는 치료용 조성물 | |
CN117384790A (zh) | 一株戊糖片球菌ks5及其在制备抗炎和助睡眠食品药品中的应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |