CN111117963A - 用于降低麻醉剂对神经细胞的影响的组合物 - Google Patents
用于降低麻醉剂对神经细胞的影响的组合物 Download PDFInfo
- Publication number
- CN111117963A CN111117963A CN201911396139.1A CN201911396139A CN111117963A CN 111117963 A CN111117963 A CN 111117963A CN 201911396139 A CN201911396139 A CN 201911396139A CN 111117963 A CN111117963 A CN 111117963A
- Authority
- CN
- China
- Prior art keywords
- percent
- fermentation
- anesthetic
- reducing
- nerve cells
- 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.)
- Granted
Links
- 210000002569 neuron Anatomy 0.000 title claims abstract description 18
- 239000000203 mixture Substances 0.000 title claims abstract description 17
- 230000000694 effects Effects 0.000 title claims description 19
- 239000003193 general anesthetic agent Substances 0.000 title description 4
- 229940035674 anesthetics Drugs 0.000 title description 2
- 238000000855 fermentation Methods 0.000 claims abstract description 45
- 230000004151 fermentation Effects 0.000 claims abstract description 45
- 241000192130 Leuconostoc mesenteroides Species 0.000 claims abstract description 29
- 230000003444 anaesthetic effect Effects 0.000 claims abstract description 20
- 239000001963 growth medium Substances 0.000 claims abstract description 19
- 239000006228 supernatant Substances 0.000 claims abstract description 13
- 239000000047 product Substances 0.000 claims abstract description 10
- 238000011146 sterile filtration Methods 0.000 claims abstract description 10
- WPYMKLBDIGXBTP-UHFFFAOYSA-N benzoic acid group Chemical group C(C1=CC=CC=C1)(=O)O WPYMKLBDIGXBTP-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000007796 conventional method Methods 0.000 claims abstract description 5
- 229920002307 Dextran Polymers 0.000 claims description 22
- 240000008042 Zea mays Species 0.000 claims description 8
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 8
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 8
- 235000005822 corn Nutrition 0.000 claims description 8
- 238000012258 culturing Methods 0.000 claims description 8
- 239000012153 distilled water Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 238000001994 activation Methods 0.000 claims description 6
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 4
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 4
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 4
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 4
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 4
- 230000003213 activating effect Effects 0.000 claims description 4
- 230000004913 activation Effects 0.000 claims description 4
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 4
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 4
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 4
- 238000009630 liquid culture Methods 0.000 claims description 4
- 239000002609 medium Substances 0.000 claims description 4
- 239000012528 membrane Substances 0.000 claims description 4
- 229920000136 polysorbate Polymers 0.000 claims description 4
- 239000011148 porous material Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000011218 seed culture Methods 0.000 claims description 4
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 claims description 4
- 239000012137 tryptone Substances 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 239000007788 liquid Substances 0.000 abstract description 7
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 11
- 210000004027 cell Anatomy 0.000 description 8
- 229960002725 isoflurane Drugs 0.000 description 8
- 210000001178 neural stem cell Anatomy 0.000 description 8
- 206010029350 Neurotoxicity Diseases 0.000 description 6
- 206010044221 Toxic encephalopathy Diseases 0.000 description 6
- 230000005764 inhibitory process Effects 0.000 description 6
- 230000007135 neurotoxicity Effects 0.000 description 6
- 231100000228 neurotoxicity Toxicity 0.000 description 6
- 230000004069 differentiation Effects 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 239000012530 fluid Substances 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 230000035755 proliferation Effects 0.000 description 3
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- 241001018563 Nekemias grossedentata Species 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 230000029087 digestion Effects 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 230000006698 induction Effects 0.000 description 2
- 238000000034 method Methods 0.000 description 2
- 210000000653 nervous system Anatomy 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 238000012353 t test Methods 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 206010006002 Bone pain Diseases 0.000 description 1
- 241000345998 Calamus manan Species 0.000 description 1
- 206010007027 Calculus urinary Diseases 0.000 description 1
- 102000009024 Epidermal Growth Factor Human genes 0.000 description 1
- 101800003838 Epidermal growth factor Proteins 0.000 description 1
- 102000003974 Fibroblast growth factor 2 Human genes 0.000 description 1
- 108090000379 Fibroblast growth factor 2 Proteins 0.000 description 1
- 208000015817 Infant Nutrition disease Diseases 0.000 description 1
- 206010061218 Inflammation Diseases 0.000 description 1
- 241000208949 Malpighiaceae Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000008730 Nestin Human genes 0.000 description 1
- 108010088225 Nestin Proteins 0.000 description 1
- 208000025747 Rheumatic disease Diseases 0.000 description 1
- 241000231739 Rutilus rutilus Species 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241001330502 Stephania Species 0.000 description 1
- 238000000692 Student's t-test Methods 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 208000009911 Urinary Calculi Diseases 0.000 description 1
- 238000002835 absorbance Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 210000001130 astrocyte Anatomy 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000004641 brain development Effects 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000002771 cell marker Substances 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000003920 cognitive function Effects 0.000 description 1
- 230000003931 cognitive performance Effects 0.000 description 1
- 201000003146 cystitis Diseases 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 210000001947 dentate gyrus Anatomy 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 230000003467 diminishing effect Effects 0.000 description 1
- 229940116977 epidermal growth factor Drugs 0.000 description 1
- 210000003754 fetus Anatomy 0.000 description 1
- 230000000971 hippocampal effect Effects 0.000 description 1
- 210000001320 hippocampus Anatomy 0.000 description 1
- 230000004054 inflammatory process Effects 0.000 description 1
- 230000013016 learning Effects 0.000 description 1
- 230000015654 memory Effects 0.000 description 1
- 230000027939 micturition Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000004081 narcotic agent Substances 0.000 description 1
- 230000003533 narcotic effect Effects 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 230000001537 neural effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 210000004248 oligodendroglia Anatomy 0.000 description 1
- 239000000902 placebo Substances 0.000 description 1
- 229940068196 placebo Drugs 0.000 description 1
- 108010055896 polyornithine Proteins 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000012950 rattan cane Nutrition 0.000 description 1
- 230000000552 rheumatic effect Effects 0.000 description 1
- 229960002078 sevoflurane Drugs 0.000 description 1
- DFEYYRMXOJXZRJ-UHFFFAOYSA-N sevoflurane Chemical compound FCOC(C(F)(F)F)C(F)(F)F DFEYYRMXOJXZRJ-UHFFFAOYSA-N 0.000 description 1
- 238000007619 statistical method Methods 0.000 description 1
- 239000004575 stone Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 208000019206 urinary tract infection Diseases 0.000 description 1
- VBEQCZHXXJYVRD-GACYYNSASA-N uroanthelone Chemical compound C([C@@H](C(=O)N[C@H](C(=O)N[C@@H](CS)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CS)C(=O)N[C@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)NCC(=O)N[C@@H](CC=1C=CC(O)=CC=1)C(=O)N[C@@H](CO)C(=O)NCC(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CS)C(=O)N[C@@H](CCC(N)=O)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCNC(N)=N)C(O)=O)C(C)C)[C@@H](C)O)NC(=O)[C@H](CO)NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@@H](NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H](CCSC)NC(=O)[C@H](CS)NC(=O)[C@@H](NC(=O)CNC(=O)CNC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CS)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)CNC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@H]1N(CCC1)C(=O)[C@H](CS)NC(=O)CNC(=O)[C@H]1N(CCC1)C(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@@H](N)CC(N)=O)C(C)C)[C@@H](C)CC)C1=CC=C(O)C=C1 VBEQCZHXXJYVRD-GACYYNSASA-N 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
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0618—Cells of the nervous system
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5014—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing toxicity
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/30—Organic components
-
- 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
- C12N2500/00—Specific components of cell culture medium
- C12N2500/70—Undefined extracts
- C12N2500/72—Undefined extracts from bacteria
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biotechnology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Immunology (AREA)
- General Engineering & Computer Science (AREA)
- Toxicology (AREA)
- Urology & Nephrology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Cell Biology (AREA)
- Hematology (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Food Science & Technology (AREA)
- Virology (AREA)
- Analytical Chemistry (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Neurology (AREA)
- Neurosurgery (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
本发明公开了一种用于降低麻醉剂对神经细胞的影响的组合物，其包括：3,4‑二羟基苯甲酸‑3‑O‑α‑L‑鼠李糖苷。其还可以进一步包括肠膜明串珠菌葡聚糖亚种LZ1发酵提取液。所述肠膜明串珠菌葡聚糖亚种LZ1发酵提取液的制备方法为：(1)将菌株肠膜明串珠菌葡聚糖亚种LZ1按照常规方法活化培养；(2)发酵：取10％种子液接种至含有3,4‑二羟基苯甲酸‑3‑O‑α‑L‑鼠李糖苷的发酵培养基中；(3)制备发酵提取物：将发酵产物离得到上清液，将所述上清液进行无菌过滤，得到肠膜明串珠菌葡聚糖亚种LZ1提取液。
Description
技术领域
本发明涉及一种用于降低麻醉剂对神经细胞的影响的组合物，属于生物技术领域。
背景技术
婴幼儿手术常采用吸入麻醉方法进行麻醉，其具有诱导快、操作简单等优点，如七氟醚、异氟醚都是常用的全麻药。吸入麻醉药对胎儿、婴幼儿大脑发育的影响，仍是目前临床关注的问题。现有技术研究表明异氟醚能够抑制新生大鼠海马齿状回神经干细胞的增殖，并影响神经干细胞向神经元的分化。异氟醚对人类神经干细胞分化以及神经形成等产生一定的影响。在动物实验研究中，人们发现达到临床手术要求的麻醉药物浓度将影响脑的学习、记忆、认知功能及行为表现能力。如何降低麻醉药对神经系统的影响是非常值得探索的课题。
神经干细胞是一类能够不断增殖、具有多向分化潜能的细胞群，其可以分化为神经组织各类细胞，如神经元、星形胶质细胞和少突胶质细胞等，周围外环境可对其分裂增殖以及分化等产生显著影响。通常使用神经干细胞来评估物质对神经系统的影响。
微生物发酵产品作为研究前沿，具有越来越重要的作用，本发明旨在提供一种用于降低麻醉剂对神经细胞的影响的组合物，具体而言，是一种发酵提取物，更进一步地，本发明还对发酵过程进行了优化以期达到更好的效果。
发明内容
本发明的目的是提供一种用于降低麻醉剂对神经细胞的影响的组合物，具体而言，是一种发酵提取物，更进一步地，本发明还对发酵过程进行了优化以期达到更好的效果。
倒心盾翅藤.系金虎尾科(Malpighiaceae)植物倒心盾翅藤的干燥藤茎，别名吼盖贯，主要分布于我国的海南、云南等地。其味淡、涩，性微寒，入膀胱经。该药作为民族用药具有消炎利尿、清热排石之效，可用于治疗尿路感染、膀胱炎，风湿骨痛，产后体虚，小儿疳积等症，是傣医传统经方“五淋化石胶囊”中的主要组成药材之一，具有较高的研究价值。现有技术(中草药Chinese Traditional and Herbal Drugs第50卷第5期2019年)有人从倒心盾翅藤中分离得到一个鼠李糖苷化合物：3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷，其分子式为C13H16O8。
肠膜明串珠菌被公认为无毒的益生菌，现有技术披露过一株肠膜明串珠菌葡聚糖亚种LZ1(南方农业学报Journal of Southern Agriculture 2019，50(1)：137-143)，其已知功能为发酵荔枝果汁。
本发明人发现，3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷可以显著降低麻醉药的神经毒性，而将其应用于肠膜明串珠菌葡聚糖亚种LZ1发酵后，其发酵提取液具有更好的降低麻醉药神经毒性的作用。
本发明所需要解决的技术问题可以通过以下技术方案来实现。
一种用于降低麻醉剂对神经细胞的影响的组合物，其包括：
3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷。
作为优选，其进一步包括肠膜明串珠菌葡聚糖亚种LZ1发酵提取液。
一种用于降低麻醉剂对神经细胞的影响的组合物的制备方法，其步骤如下：
(1)将菌株肠膜明串珠菌葡聚糖亚种LZ1按照常规方法活化培养；
作为优选，其活化过程为：用接种环取保藏好的菌，接种到MRS液体培养基中活化，然后将单个克隆接种到种子培养基中，30℃培养24小时，制备种子液，得到5.0*107(CFU/ml)肠膜明串珠菌葡聚糖亚种LZ1种子液；
(2)发酵：
取10％种子液接种至发酵培养基中；
所述发酵培养基配方：按照质量分数，3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷0.8％，酵母粉2％、胰蛋白胨6％、麦芽糖6％、海藻糖5％、柠檬酸铵1％、玉米浸汁(玉米浆)9％、K2HPO41％，(NH4)2SO40.3％，吐温0.3％，其余为蒸馏水；
培养条件为：温度38℃，培养时间为：24hr得到发酵产物；
(3)制备发酵提取物：
将发酵产物于1800rpm离心20分钟，得到上清液，将所述上清液进行无菌过滤，选择孔径0.22μm的滤膜进行无菌过滤，得到肠膜明串珠菌葡聚糖亚种LZ1提取液。
本发明的优点：
本发明人发现，3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷可以显著降低麻醉药的神经毒性，而将其应用于肠膜明串珠菌葡聚糖亚种LZ1发酵后，其发酵提取液具有更好的降低麻醉药神经毒性的作用，具有非常广阔的应用前景。
具体实施方式
下面详细描述本发明的实施例，所述实施例仅用于解释本发明，而不能理解为对本发明的限制。
本发明的具体实施例如以下说明。
本实施例所用肠膜明串珠菌葡聚糖亚种LZ1为赠送得到。
实施例1
配置2组干预组合物：
组1为单纯3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷：
将3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷溶于DMF后，加入生理盐水，制备成终浓度为0.8％的3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷溶液。
组2为肠膜明串珠菌葡聚糖亚种LZ1的3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷干预发酵提取液：
(1)将菌株肠膜明串珠菌葡聚糖亚种LZ1按照常规方法活化培养；
用接种环取保藏好的菌，接种到MRS液体培养基中活化，然后将单个克隆接种到种子培养基中，30℃培养24小时，制备种子液，得到5.0*107(CFU/ml)肠膜明串珠菌葡聚糖亚种LZ1种子液；
(2)发酵：
取10％种子液接种至发酵培养基中；
所述发酵培养基配方：按照质量分数，3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷0.8％，酵母粉2％、胰蛋白胨6％、麦芽糖6％、海藻糖5％、柠檬酸铵1％、玉米浸汁(玉米浆)9％、K2HPO41％，(NH4)2SO40.3％，吐温0.3％，其余为蒸馏水；
培养条件为：温度38℃，培养时间为：24hr得到发酵产物；
(3)制备发酵提取物：
将发酵产物于1800rpm离心20分钟，得到上清液，将所述上清液进行无菌过滤，选择孔径0.22μm的滤膜进行无菌过滤，得到肠膜明串珠菌葡聚糖亚种LZ1提取液。
组3为肠膜明串珠菌葡聚糖亚种LZ1常规发酵提取液：
(1)将菌株肠膜明串珠菌葡聚糖亚种LZ1按照常规方法活化培养；
用接种环取保藏好的菌，接种到MRS液体培养基中活化，然后将单个克隆接种到种子培养基中，30℃培养24小时，制备种子液，得到5.0*107(CFU/ml)肠膜明串珠菌葡聚糖亚种LZ1种子液；
(2)发酵：
取10％种子液接种至发酵培养基中；
所述发酵培养基配方：酵母粉2％、胰蛋白胨6％、麦芽糖6％、海藻糖5％、柠檬酸铵1％、玉米浸汁(玉米浆)9％、K2HPO41％，(NH4)2SO40.3％，吐温0.3％，其余为蒸馏水；
培养条件为：温度38℃，培养时间为：24hr得到发酵产物；
(3)制备发酵提取物：
将发酵产物于1800rpm离心20分钟，得到上清液，将所述上清液进行无菌过滤，选择孔径0.22μm的滤膜进行无菌过滤，得到肠膜明串珠菌葡聚糖亚种LZ1提取液。
实施例2
取出生后第7天SD大鼠(体质量12-16g)。无菌条件下分离出海马组织，PBS漂洗，用眼科剪剪碎成糊状，放入胰蛋白酶中消化，根据消化情况加入含体积分数为10％胎牛血清的PBS中止消化，吸管机械吹打后，1200r/min离心5min弃上清，沉淀加入无血清DMEM/F12养液(含2％B27，20ng/L表皮生长因子和20ng/L碱性成纤维细胞生长因子)混悬，调整细胞浓度为1×109/L，置于37℃，体积分数为5％CO2培养箱进行培养，每隔两三天换液1次。原代培养5-7d传代，神经干细胞标志蛋白Nestin鉴定神经干细胞，传代至第3代细胞进行实验。
将神经干细胞接种至多聚鸟氨酸和层粘连蛋白包被的培养板，加入含体积分数为1％胎牛血清、1％青链霉素、2％B27的DMEM/F12培养基进行诱导培养8d。
将6孔培养板细胞置于实验箱。
实验分为5组：对照组、异氟醚组、组1干预组、组2干预组、组3干预组。不同处理条件如下：
对照组：仅通入体积分数为5％CO2，体积分数为95％O2的混合气体；
异氟醚组：通入2.8％异氟醚和体积分数为5％CO2，体积分数为95％O2的混合气体；
组1干预组：通入2.8％异氟醚和体积分数为5％CO2，体积分数为95％O2的混合气体，按照培养基：干预液为10:1的比例加入组1的干预液；
组2干预组：通入2.8％异氟醚和体积分数为5％CO2，体积分数为95％O2的混合气体，按照培养基：干预液为10:1的比例加入组2的干预液；
组3干预组：通入2.8％异氟醚和体积分数为5％CO2，体积分数为95％O2的混合气体，按照培养基：干预液为10:1的比例加入组3的干预液；
干预6h后，取出6孔培养板再放入37℃细胞培养箱中继续培养2h。
实施例3
取实施例2中各组细胞，随机重复3次。采用MTT法进行细胞增殖-毒性检测。使用MTT试剂盒检测细胞增殖能力，于37℃下、570nm波长处，以酶标仪测定每孔吸光度(A)。抑制率(％)＝(1－A干预组/A对照组)×100％。
统计学处理：使用SPSS16.0,进行统计学分析，计量资料的结果表示为均值±标准差,采用组间t检验。
测定实施例2的各处理组的细胞增殖情况，结果如表1所示：
表1不同组别的细胞增殖情况
注：t检验,*：P＜0.05(与空白对照组比较)；#：P＜0.05(和异氟醚组比较)；
如上结果可见，异氟醚组的抑制率最高，组2干预组的抑制率最低。组1干预组的抑制率有所减少；组2干预组和空白对照组的抑制率无统计学差异(P＞0.05)；组3干预组和空白对照有统计学差异(P＜0.05)，但是和异氟醚组的抑制率无统计学差异，显示肠膜明串珠菌葡聚糖亚种LZ1常规发酵提取液效果欠佳，3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷可以显著降低麻醉药的神经毒性，其应用于肠膜明串珠菌葡聚糖亚种LZ1发酵后的提取液具有更好的降低麻醉药神经毒性的作用。
需要说明的是，以上所述仅为本发明的优选具体的实施例，
若依本发明的构想所作变动，其产生的功能作用，仍未超出说明书所涵盖的精神时，均应在本发明的范围内。
在本说明书的描述中，参考术语“一个实施例”、“一些实施例”、“示例”、“具体示例”、或“一些示例”等的描述意指结合该实施例或示例描述的具体特征、结构、材料或者特点包含于本发明的至少一个实施例或示例中。在本说明书中，对上述术语的示意性表述不一定指的是相同的实施例或示例。而且，描述的具体特征、结构、材料或者特点可以在任何的一个或多个实施例或示例中以合适的方式结合。
尽管已经示出和描述了本发明的实施例，本领域的普通技术人员可以理解：在不脱离本发明的原理和宗旨的情况下可以对这些实施例进行多种变化、修改、替换和变型，本发明的范围由权利要求及其等同物限定。
Claims (8)
1.一种用于降低麻醉剂对神经细胞的影响的组合物，其包括：
3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷。
2.权利要求1所述的降低麻醉剂对神经细胞的影响的组合物，其特征在于：
其进一步包括肠膜明串珠菌葡聚糖亚种LZ1发酵提取液。
3.权利要求2所述的降低麻醉剂对神经细胞的影响的组合物，其特征在于：
所述肠膜明串珠菌葡聚糖亚种LZ1发酵提取液的制备方法为：
(1)将菌株肠膜明串珠菌葡聚糖亚种LZ1按照常规方法活化培养；
(2)发酵：
取10％种子液接种至含有3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷的发酵培养基中；
(3)制备发酵提取物：
将发酵产物离得到上清液，将所述上清液进行无菌过滤，得到肠膜明串珠菌葡聚糖亚种LZ1提取液。
4.权利要求3所述的降低麻醉剂对神经细胞的影响的组合物，其特征在于：
步骤(1)活化过程为：用接种环取保藏好的菌，接种到MRS液体培养基中活化，然后将单个克隆接种到种子培养基中，30℃培养24小时，制备种子液，得到5.0*107(CFU/ml)肠膜明串珠菌葡聚糖亚种LZ1种子液。
5.权利要求3所述的降低麻醉剂对神经细胞的影响的组合物，其特征在于：
所述发酵培养基配方：按照质量分数，3,4-二羟基苯甲酸-3-O-α-L-鼠李糖苷0.8％，酵母粉2％、胰蛋白胨6％、麦芽糖6％、海藻糖5％、柠檬酸铵1％、玉米浸汁(玉米浆)9％、K2HPO41％，(NH4)2SO40.3％，吐温0.3％，其余为蒸馏水；
培养条件为：温度38℃，培养时间为：24hr得到发酵产物。
6.权利要求3所述的降低麻醉剂对神经细胞的影响的组合物，其特征在于：
步骤(3)为：将发酵产物于1800rpm离心20分钟，得到上清液，将所述上清液进行无菌过滤，选择孔径0.22μm的滤膜进行无菌过滤，得到肠膜明串珠菌葡聚糖亚种LZ1提取液。
7.权利要求1-6任一项所述的降低麻醉剂对神经细胞的影响的组合物在制药上的应用。
8.制备权利要求1-6任一项所述的降低麻醉剂对神经细胞的影响的组合物的方法。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911396139.1A CN111117963B (zh) | 2019-12-30 | 2019-12-30 | 组合物及其在神经干细胞培养上的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911396139.1A CN111117963B (zh) | 2019-12-30 | 2019-12-30 | 组合物及其在神经干细胞培养上的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111117963A true CN111117963A (zh) | 2020-05-08 |
| CN111117963B CN111117963B (zh) | 2021-08-27 |
Family
ID=70505144
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911396139.1A Active CN111117963B (zh) | 2019-12-30 | 2019-12-30 | 组合物及其在神经干细胞培养上的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111117963B (zh) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102453735A (zh) * | 2010-10-15 | 2012-05-16 | 许明淑 | 采用柚苷酶从淫羊藿苷获得淫羊藿苷元的方法 |
| CN103320339A (zh) * | 2013-03-12 | 2013-09-25 | 东莞市农业科学研究中心 | 一株肠膜明串珠菌菌株 |
| CN104004714A (zh) * | 2014-04-30 | 2014-08-27 | 南通大学 | 谷氨酸诱导神经干细胞向神经元分化的方法 |
| CN104830898A (zh) * | 2015-05-15 | 2015-08-12 | 湖南工程学院 | 一种提高人参根中人参皂苷Rg1含量的方法与应用 |
| WO2019246282A1 (en) * | 2018-06-19 | 2019-12-26 | The Regents Of The University Of Colorado | Protection from stress, anxiety, neuroinflammation and cognitive dysfunction |
-
2019
- 2019-12-30 CN CN201911396139.1A patent/CN111117963B/zh active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102453735A (zh) * | 2010-10-15 | 2012-05-16 | 许明淑 | 采用柚苷酶从淫羊藿苷获得淫羊藿苷元的方法 |
| CN103320339A (zh) * | 2013-03-12 | 2013-09-25 | 东莞市农业科学研究中心 | 一株肠膜明串珠菌菌株 |
| CN103320339B (zh) * | 2013-03-12 | 2015-03-11 | 东莞市农业科学研究中心 | 一株肠膜明串珠菌菌株 |
| CN104004714A (zh) * | 2014-04-30 | 2014-08-27 | 南通大学 | 谷氨酸诱导神经干细胞向神经元分化的方法 |
| CN104830898A (zh) * | 2015-05-15 | 2015-08-12 | 湖南工程学院 | 一种提高人参根中人参皂苷Rg1含量的方法与应用 |
| WO2019246282A1 (en) * | 2018-06-19 | 2019-12-26 | The Regents Of The University Of Colorado | Protection from stress, anxiety, neuroinflammation and cognitive dysfunction |
Non-Patent Citations (1)
| Title |
|---|
| 赵序利等: "异氟醚通过激活Ryanodine受体对ReNcell CX人神经干细胞损伤和存活发挥双向调节作用 ", 《中国医药科学》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111117963B (zh) | 2021-08-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108403498B (zh) | 一种纳豆提取液的制备方法及其应用 | |
| KR20140063948A (ko) | 진세노사이드 Rg3를 증진하고 항산화 기능을 촉진하기 위한 발효 홍삼 농축액 제조 방법 | |
| CN105193876A (zh) | 一种马齿苋提取物及其制备方法 | |
| TWI711458B (zh) | 植物發酵物及其製備方法與用於胃臟保健的用途 | |
| CN114652748A (zh) | 一种含有干细胞抑菌因子的医用妇科洗液的制备方法及其应用 | |
| EP2116252A1 (en) | The use of epimedium flavones and effective components thereof for the preparation of medicaments of promoting proliferations and differentiations of nerve cells | |
| CN105687036A (zh) | 白及护肤产品及其制备方法 | |
| CN111117963B (zh) | 组合物及其在神经干细胞培养上的应用 | |
| CN116509905A (zh) | 一种具有缓解便秘作用的两歧双歧杆菌生物制剂 | |
| CN116549535A (zh) | 降尿酸的发酵物及其制备方法和用途 | |
| CN115887535B (zh) | 一种防治禽大肠杆菌病的复方中药渣发酵液及其制备方法 | |
| CN107468715A (zh) | 蜡状芽孢杆菌cgmcc0601发酵液在制备治疗糖尿病创伤伤口药物中的应用 | |
| CN106755256A (zh) | 一种土鳖虫多肽的制备方法、制备得到的土鳖虫多肽及其应用 | |
| CN110787281A (zh) | 治疗皮肤组织溃疡感染的抗菌肽复方植物药凝胶及制备方法 | |
| CN107375480A (zh) | 大承气汤在制备防治帕金森氏病药物中的应用 | |
| CN111494306A (zh) | 一种女性私密凝胶微生态制剂组合物及制备方法 | |
| CN103520396B (zh) | 一种用于预防口腔种植修复后感染的漱口液 | |
| CN110420241A (zh) | 一种治疗前列腺炎的中药喷膜剂及其制备方法 | |
| CN115386523B (zh) | 一株乳酸乳球菌及其在抗幽门螺杆菌感染的应用 | |
| CN110575459A (zh) | 粪菌移植胶囊的制备方法 | |
| CN107375479A (zh) | 大承气汤在制备改善由中枢炎症造成的老年痴呆药物中的应用 | |
| CN115569165B (zh) | 中药组合物在制备治疗细菌性前列腺炎的药物中的应用 | |
| CN116656609A (zh) | 一种骨髓神经嵴细胞分泌的胞外囊泡的制备方法及用途 | |
| CN117503800B (zh) | 一种胃黏膜上皮细胞提取液及其制备方法与应用 | |
| CN110938599B (zh) | 神经干细胞的培养方法及其应用 |
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 | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20200729 Address after: No. 1800 road 214000 Jiangsu Lihu Binhu District City of Wuxi Province Applicant after: Jiangnan University Applicant after: THE FIRST AFFILIATED HOSPITAL OF HEBEI NORTH University Address before: 075061 No. 12, Changqing Road, Qiaoxi West, Hebei, Zhangjiakou Applicant before: THE FIRST AFFILIATED HOSPITAL OF HEBEI NORTH University |
|
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20210224 Address after: 075000 No. 12, Changqing Road, Qiaoxi West, Hebei, Zhangjiakou Applicant after: THE FIRST AFFILIATED HOSPITAL OF HEBEI NORTH University Address before: 214000 1800 Lihu Avenue, Binhu District, Wuxi, Jiangsu Applicant before: Jiangnan University Applicant before: THE FIRST AFFILIATED HOSPITAL OF HEBEI NORTH University |
|
| TA01 | Transfer of patent application right | ||
| CB03 | Change of inventor or designer information |
Inventor after: Wang Taihua Inventor after: Shi Xinyi Inventor after: Yao Jie Inventor before: Yao Jie |
|
| CB03 | Change of inventor or designer information | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20210810 Address after: 523000 401, room 4, building 10, innovation and Technology Park, Songshan Lake high tech Industrial Development Zone, Dongguan, Guangdong, China Applicant after: Guangdong cel Biotechnology Co.,Ltd. Address before: 075000 No. 12, Changqing Road, Qiaoxi West, Hebei, Zhangjiakou Applicant before: THE FIRST AFFILIATED HOSPITAL OF HEBEI NORTH University |
|
| TA01 | Transfer of patent application right | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP02 | Change in the address of a patent holder |
Address after: Room 401, Building 10, No. 6, Hongmian Road, Songshan Lake Park, Dongguan City, Guangdong 523000 Patentee after: Guangdong cel Biotechnology Co.,Ltd. Address before: 523000 401, room 4, building 10, innovation and Technology Park, Songshan Lake high tech Industrial Development Zone, Dongguan, Guangdong, China Patentee before: Guangdong cel Biotechnology Co.,Ltd. |
|
| CP02 | Change in the address of a patent holder |