CN113181229A - Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 - Google Patents
Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 Download PDFInfo
- Publication number
- CN113181229A CN113181229A CN202110478468.1A CN202110478468A CN113181229A CN 113181229 A CN113181229 A CN 113181229A CN 202110478468 A CN202110478468 A CN 202110478468A CN 113181229 A CN113181229 A CN 113181229A
- Authority
- CN
- China
- Prior art keywords
- cov
- sars
- isatis tinctoria
- cabbage type
- type rape
- 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
- 239000000178 monomer Substances 0.000 title claims abstract description 79
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 title claims abstract description 44
- 240000007124 Brassica oleracea Species 0.000 title claims abstract description 43
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 title claims abstract description 43
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 title claims abstract description 43
- 241001678559 COVID-19 virus Species 0.000 title claims abstract description 7
- 230000002401 inhibitory effect Effects 0.000 title claims description 11
- 239000000284 extract Substances 0.000 claims abstract description 43
- 208000015181 infectious disease Diseases 0.000 claims abstract description 17
- 238000002360 preparation method Methods 0.000 claims abstract description 16
- 238000000605 extraction Methods 0.000 claims abstract description 14
- 241000711573 Coronaviridae Species 0.000 claims description 43
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 claims description 39
- 241000700605 Viruses Species 0.000 claims description 32
- 239000003814 drug Substances 0.000 claims description 32
- 241000699670 Mus sp. Species 0.000 claims description 30
- 235000011331 Brassica Nutrition 0.000 claims description 25
- 241000219198 Brassica Species 0.000 claims description 25
- 208000025721 COVID-19 Diseases 0.000 claims description 19
- 208000037847 SARS-CoV-2-infection Diseases 0.000 claims description 18
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 16
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 claims description 15
- 210000004072 lung Anatomy 0.000 claims description 13
- 230000004083 survival effect Effects 0.000 claims description 13
- 229940079593 drug Drugs 0.000 claims description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 11
- 238000001704 evaporation Methods 0.000 claims description 8
- 239000000287 crude extract Substances 0.000 claims description 5
- 238000000034 method Methods 0.000 claims description 5
- 238000004321 preservation Methods 0.000 claims description 5
- 239000006228 supernatant Substances 0.000 claims description 5
- 239000012153 distilled water Substances 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000009210 therapy by ultrasound Methods 0.000 claims description 4
- 239000013585 weight reducing agent Substances 0.000 claims description 4
- 239000000654 additive Substances 0.000 claims description 3
- 230000000996 additive effect Effects 0.000 claims description 3
- 238000007877 drug screening Methods 0.000 claims description 2
- 238000002156 mixing Methods 0.000 claims description 2
- 238000012216 screening Methods 0.000 claims description 2
- 229940126585 therapeutic drug Drugs 0.000 claims description 2
- 230000001225 therapeutic effect Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 33
- 241001465754 Metazoa Species 0.000 abstract description 10
- 230000000840 anti-viral effect Effects 0.000 abstract description 8
- 230000008901 benefit Effects 0.000 abstract description 7
- 230000005764 inhibitory process Effects 0.000 abstract description 7
- 238000011161 development Methods 0.000 abstract description 5
- 230000036541 health Effects 0.000 abstract description 5
- 231100000331 toxic Toxicity 0.000 abstract description 5
- 230000002588 toxic effect Effects 0.000 abstract description 5
- 235000013311 vegetables Nutrition 0.000 abstract description 4
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 239000002994 raw material Substances 0.000 abstract description 3
- 230000002900 effect on cell Effects 0.000 abstract description 2
- 235000013305 food Nutrition 0.000 abstract description 2
- 210000004027 cell Anatomy 0.000 description 39
- 241000334154 Isatis tinctoria Species 0.000 description 23
- 240000002791 Brassica napus Species 0.000 description 12
- 102100028279 WW domain-binding protein 1 Human genes 0.000 description 11
- 101710160563 WW domain-binding protein 1 Proteins 0.000 description 11
- 235000011293 Brassica napus Nutrition 0.000 description 10
- 241000334160 Isatis Species 0.000 description 9
- 239000012528 membrane Substances 0.000 description 9
- 230000035755 proliferation Effects 0.000 description 8
- 201000003176 Severe Acute Respiratory Syndrome Diseases 0.000 description 7
- 230000037396 body weight Effects 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 210000001519 tissue Anatomy 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 6
- 239000010410 layer Substances 0.000 description 6
- 238000001035 drying Methods 0.000 description 5
- 238000000338 in vitro Methods 0.000 description 5
- 238000001727 in vivo Methods 0.000 description 5
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 4
- 206010035664 Pneumonia Diseases 0.000 description 4
- 241000315672 SARS coronavirus Species 0.000 description 4
- 241001557415 Tetrastigma hemsleyanum Species 0.000 description 4
- 238000012258 culturing Methods 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 238000011534 incubation Methods 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000007789 sealing Methods 0.000 description 4
- 239000000243 solution Substances 0.000 description 4
- 241000712461 unidentified influenza virus Species 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- 241000008904 Betacoronavirus Species 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 239000013543 active substance Substances 0.000 description 3
- 239000003443 antiviral agent Substances 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 230000001413 cellular effect Effects 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 230000003247 decreasing effect Effects 0.000 description 3
- 238000001962 electrophoresis Methods 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 238000010874 in vitro model Methods 0.000 description 3
- 238000002386 leaching Methods 0.000 description 3
- 239000002609 medium Substances 0.000 description 3
- 239000000419 plant extract Substances 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 238000011725 BALB/c mouse Methods 0.000 description 2
- 235000006008 Brassica napus var napus Nutrition 0.000 description 2
- 206010058467 Lung neoplasm malignant Diseases 0.000 description 2
- 239000006180 TBST buffer Substances 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000009835 boiling Methods 0.000 description 2
- 239000006143 cell culture medium Substances 0.000 description 2
- 210000000349 chromosome Anatomy 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 238000003745 diagnosis Methods 0.000 description 2
- 239000000469 ethanolic extract Substances 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 239000012894 fetal calf serum Substances 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 241000411851 herbal medicine Species 0.000 description 2
- 210000004754 hybrid cell Anatomy 0.000 description 2
- 238000003119 immunoblot Methods 0.000 description 2
- 230000007365 immunoregulation Effects 0.000 description 2
- 231100000518 lethal Toxicity 0.000 description 2
- 230000001665 lethal effect Effects 0.000 description 2
- 201000005202 lung cancer Diseases 0.000 description 2
- 208000020816 lung neoplasm Diseases 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- BDAGIHXWWSANSR-UHFFFAOYSA-N methanoic acid Natural products OC=O BDAGIHXWWSANSR-UHFFFAOYSA-N 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 239000002904 solvent Substances 0.000 description 2
- 210000001082 somatic cell Anatomy 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
- 239000000126 substance Substances 0.000 description 2
- 229940126680 traditional chinese medicines Drugs 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 230000005727 virus proliferation Effects 0.000 description 2
- OSWFIVFLDKOXQC-UHFFFAOYSA-N 4-(3-methoxyphenyl)aniline Chemical compound COC1=CC=CC(C=2C=CC(N)=CC=2)=C1 OSWFIVFLDKOXQC-UHFFFAOYSA-N 0.000 description 1
- 235000004977 Brassica sinapistrum Nutrition 0.000 description 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 description 1
- 241001674939 Caulanthus Species 0.000 description 1
- 206010009944 Colon cancer Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 206010069767 H1N1 influenza Diseases 0.000 description 1
- 241000127282 Middle East respiratory syndrome-related coronavirus Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 241000712464 Orthomyxoviridae Species 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000004952 Polyamide Substances 0.000 description 1
- 206010037368 Pulmonary congestion Diseases 0.000 description 1
- 108091006197 SARS-CoV-2 Nucleocapsid Protein Proteins 0.000 description 1
- 208000036142 Viral infection Diseases 0.000 description 1
- 229960005305 adenosine Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000000844 anti-bacterial effect Effects 0.000 description 1
- 230000001093 anti-cancer Effects 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 230000003833 cell viability Effects 0.000 description 1
- 208000029742 colonic neoplasm Diseases 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000002380 cytological effect Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 230000000857 drug effect Effects 0.000 description 1
- 230000000312 effect on influenza Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000013210 evaluation model Methods 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 235000019253 formic acid Nutrition 0.000 description 1
- 229940013688 formic acid Drugs 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 238000012252 genetic analysis Methods 0.000 description 1
- 238000012214 genetic breeding Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 150000004676 glycans Chemical class 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 238000007489 histopathology method Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 210000004969 inflammatory cell Anatomy 0.000 description 1
- 238000011031 large-scale manufacturing process Methods 0.000 description 1
- 231100000225 lethality Toxicity 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000009456 molecular mechanism Effects 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- VSZGPKBBMSAYNT-RRFJBIMHSA-N oseltamivir Chemical compound CCOC(=O)C1=C[C@@H](OC(CC)CC)[C@H](NC(C)=O)[C@@H](N)C1 VSZGPKBBMSAYNT-RRFJBIMHSA-N 0.000 description 1
- 229960003752 oseltamivir Drugs 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- XRQDFNLINLXZLB-CKIKVBCHSA-N peramivir Chemical compound CCC(CC)[C@H](NC(C)=O)[C@@H]1[C@H](O)[C@@H](C(O)=O)C[C@H]1NC(N)=N XRQDFNLINLXZLB-CKIKVBCHSA-N 0.000 description 1
- 229960001084 peramivir Drugs 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229920002647 polyamide Polymers 0.000 description 1
- 229920001282 polysaccharide Polymers 0.000 description 1
- 239000005017 polysaccharide Substances 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000029058 respiratory gaseous exchange Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000002390 rotary evaporation Methods 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 238000007873 sieving Methods 0.000 description 1
- 102000030938 small GTPase Human genes 0.000 description 1
- 108060007624 small GTPase Proteins 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 201000010740 swine influenza Diseases 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 201000008827 tuberculosis Diseases 0.000 description 1
- 230000006394 virus-host interaction Effects 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 229960001028 zanamivir Drugs 0.000 description 1
- ARAIBEBZBOPLMB-UFGQHTETSA-N zanamivir Chemical compound CC(=O)N[C@@H]1[C@@H](N=C(N)N)C=C(C(O)=O)O[C@H]1[C@H](O)[C@H](O)CO ARAIBEBZBOPLMB-UFGQHTETSA-N 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
- A61K36/315—Isatis, e.g. Dyer's woad
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/30—Animal feeding-stuffs from material of plant origin, e.g. roots, seeds or hay; from material of fungal origin, e.g. mushrooms
-
- 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
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/31—Brassicaceae or Cruciferae (Mustard family), e.g. broccoli, cabbage or kohlrabi
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
- A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
- A61P31/12—Antivirals
- A61P31/14—Antivirals for RNA viruses
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Botany (AREA)
- Medicinal Chemistry (AREA)
- Polymers & Plastics (AREA)
- Epidemiology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Zoology (AREA)
- Food Science & Technology (AREA)
- Virology (AREA)
- Alternative & Traditional Medicine (AREA)
- Molecular Biology (AREA)
- Medical Informatics (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Physiology (AREA)
- Diabetes (AREA)
- Rheumatology (AREA)
- Toxicology (AREA)
- Urology & Nephrology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pulmonology (AREA)
- Gastroenterology & Hepatology (AREA)
- Endocrinology (AREA)
- Pathology (AREA)
- Biomedical Technology (AREA)
- Animal Husbandry (AREA)
- Communicable Diseases (AREA)
Abstract
The invention belongs to the field of biotechnology, and discloses application of a cabbage type rape-isatis tinctoria G monomer addition system in inhibition of novel coronavirus SARS-CoV-2. The invention discovers for the first time that the cabbage type rape-isatis tinctoria G monomer addition line extract has obvious effect of resisting SARS-CoV-2 virus on a cell infection model and has no toxic or side effect on cells. The cabbage type rape-isatis tinctoria additional line has the advantage of large-scale planting, can provide sufficient raw materials for production and extraction of an anti-SARS-CoV-2 preparation, has low cost and high yield, can be used as antiviral medicinal and edible vegetables, feeds and the like for development and utilization, can be used as health-care food for human and animals, and can ensure the health of the animals and the human.
Description
Technical Field
The invention belongs to the field of biotechnology, and particularly relates to application of a cabbage type rape-isatis tinctoria G monomer additional system in inhibiting novel coronavirus SARS-CoV-2
Background
The novel coronavirus pneumonia (CoVID-19) is caused by severe acute respiratory syndrome coronavirus type 2 (SARS-CoV-2). However, there is no known effective new method for treating coronary pneumonia or anti-SARS-CoV-2 virus, and therefore, it is important to research and develop a novel anti-SARS-CoV-2 virus preparation or drug.
The cabbage type rape-isatis tinctoria additional line (isatis tinctoria or oil-coated isatis tinctoria for short) is an additional line formed after the cabbage type rape and isatis tinctoria cells are hybridized and fused. The rape has multiple functions and purposes, the leaves and the flower shoots can be used for vegetables, the seeds can be used for oil extraction, the rapeseed dregs and the green feed can be eaten by animals, and the rape flowers also have ornamental value. The rape is widely planted in the world, the source is wide, and the yield is high. While isatis tinctoria is an important medicinal plant in China, the root of isatis tinctoria is isatis root, the leaf of isatis tinctoria is isatis leaf, and isatis tinctoria has antibacterial, endotoxin-resisting, anticancer, immunoregulation, blood circulation-promoting, blood stasis-removing and antiviral effects, in "diagnosis and treatment scheme for pneumonia infected by novel coronavirus" (trial first to seventh edition), abbreviated as "diagnosis and treatment scheme" and 13 provinces recommend a traditional Chinese medicine treatment scheme group, isatis tinctoria is one of the medicines, but the use frequency of 18 used traditional Chinese medicines is the second to last, and is only 2 times (old and natural, etc., the characteristics and application of immunoregulation antiviral traditional Chinese medicines [ J ]. Chinese herbal medicines, volume 51, 6, 3-month 2020); in related research and news reports, the compound isatis root instead of the isatis root has the effect of resisting the new coronavirus at the in vitro cell level. Isatis tinctoria is also much lower than oilseed rape in terms of source and yield. The cabbage type rape-Isatis tinctoria additional line is obtained by hybridizing somatic cells of cabbage type rape and Isatis tinctoria, backcrossing the formed somatic cell hybrid with cabbage type rape continuously for multiple generations, and adding 7 chromosomes of Isatis tinctoria to rape respectively to obtain 7 additional lines (A, B, C, D, E, F, G). At present, the addition lines are mainly used for rape genetic breeding and analysis of isatis indigotica fort genome structure and medicinal components. Although some related research reports show that some addition systems in the brassica napus-isatis tinctoria addition system have certain antiviral effect on influenza viruses, for example, the 5 monomer addition systems (B monomer, C monomer, D monomer, E monomer and F monomer) and the disomic addition system (D doublet) are extracted by using water as a solvent in the Huang-Luciling laboratory of Chinese academy of sciences, and the D doublet is found to have certain inhibition effect on the H1N1 influenza viruses. However, there has been no report on the effect of different addition lines of Brassica napus-Isatis indigotica on the resistance to SARS-CoV-2 virus. Influenza viruses belong to the orthomyxoviridae family and SARS-CoV-2 viruses belong to the coronaviridae family. According to phylogenetic trees, coronaviruses can be divided into four genera: alpha, beta, gamma and delta, wherein the beta coronavirus can be divided into A, B, C and D four independent subgroups, MERS-CoV which is known to cause serious diseases belongs to the beta coronavirus C subgroup, SARS-CoV belongs to the beta coronavirus B subgroup, and the novel coronavirus and SARS-CoV genome have higher homology and are named as SARS-CoV-2. Lihui et al (2019 novel coronavirus antiviral treatment available drug available period [ J ]. China tuberculosis and journal of respiration, 2020, Vol 43, No 3) report that clinically common antiviral drugs including oseltamivir, peramivir, zanamivir and other drugs having a good effect on influenza viruses are ineffective against SARS-CoV-2. Although many studies on SARS-CoV-2 have been published, the current understanding of this novel coronavirus is only in Bingshan mountain corner, the pathogenesis of SARS-CoV-2 infection and the molecular mechanisms of virus-host interaction remain unclear, and there is no specific drug for treating this virus.
The drug effect of the traditional Chinese medicine is closely related to the content and the composition of active substances, and the leaching and the composition of the active substances of the traditional Chinese medicine are influenced by different extraction methods. In the invention, methanol is used as a solvent, ultrasonic treatment is carried out at 55 ℃, effective components of different addition systems are extracted by a method of rotary evaporation at 50 ℃ on a rotary evaporator, and the extracted product is dried at 50 ℃ and then used for subsequent tests. Research reports exist (Rouyun, etc., the influence of different drying processes on effective components in the isatis root water extract [ J ]. Chinese herbal medicines, No. 42, No. 8 of 2011 in 8), and different drying modes have great influence on the contents of formic acid, salicylic acid, adenosine and polysaccharide in the isatis root water extract. Li shang bin and the like research the influence of 3 extraction methods on the content of flavonoid active ingredients in the tetrastigma hemsleyanum ethanol extract and the proliferation activity of anti-lung cancer cells a549, and as a result, it is found that different extraction processes have certain influence on the leaching of the tetrastigma hemsleyanum flavonoid substances, and further influence the leaching of active substances in the tetrastigma hemsleyanum which exert a main-effect tumor inhibition effect (li shang bin and the like, influence of different extraction methods on the content of the tetrastigma hemsleyanum ethanol extract and the proliferation inhibition of lung cancer cells a549 [ J ]. zhejiang journal of traditional Chinese medicine, vol.54, No. 5, 2019).
In the invention, the extracts of different addition lines of cabbage type rape-isatis tinctoria are screened to resist SARS-CoV-2 virus, and the result shows that the extract of one addition line has obvious inhibition effect on SARS-CoV-2 virus, has obvious effect of resisting new coronavirus and relieving the damage of new coronavirus to mouse lung tissue in a mouse infection model with cell level and new coronavirus lethality, reduces weight loss caused by SARS-CoV-2 infection, and improves the survival rate of mice. With the outbreak of new coronary pneumonia, the scientific attempt of anti-viral infection and treatment to health and medical care is heavy, and the challenge of large-scale production is quickly faced when searching for anti-viral drugs, the cabbage type rape-isatis indigotica fort additional line has the advantage of large-scale planting, sufficient raw materials can be provided for production and extraction of anti-SARS-CoV-2 preparations, and meanwhile, the cabbage type rape-isatis indigotica additional line can also be developed and utilized as medicinal and edible vegetables, feeds and the like, and can be used as health food for human and animals to guarantee the health of the animals and the human.
Disclosure of Invention
The invention aims to make up the defects of the prior art and solve the problem that severe acute respiratory syndrome coronavirus (SARS-CoV-2) is difficult to prevent and treat, and provides an application of a cabbage type rape-isatis tinctoria G monomer additional system in preparing a preparation for treating or preventing novel coronavirus SARS-CoV-2 infection.
In order to achieve the purpose, the invention adopts the following technical measures:
the application of cabbage type rape-isatis tinctoria G monomer additive in preparing medicine for preventing and treating SARS-CoV-2 infection,
the application comprises the following steps:
the application of cabbage type rape-isatis tinctoria G monomer addition system in the preparation of SARS-CoV-2 inhibiting medicine;
the application of cabbage type rape-isatis tinctoria G monomer additional system in preparing medicine for preventing and treating SARS-CoV-2 infection;
the application of cabbage type rape-isatis tinctoria G monomer addition system in the preparation of medicines for relieving the weight reduction after SARS-CoV-2 infection;
the application of cabbage type rape-isatis tinctoria G monomer addition system in preparing medicine for raising the survival rate of SARS-CoV-2 infected body;
the application of cabbage type rape-isatis tinctoria G monomer additive in preparing medicine for lowering virus content in body's lung after SARS-CoV-2 infection.
The application of cabbage type rape-isatis tinctoria G monomer additional system in preparing medicine for preventing and treating mouse SARS-CoV-2 infection;
the application of cabbage type rape-isatis tinctoria G monomer additional system in preparing medicine for raising the survival rate of mouse SARS-CoV-2 infection;
the application of the cabbage type rape-isatis tinctoria G monomer additional system in preparing a medicine for relieving the weight reduction of mice after SARS-CoV-2 infection;
or the cabbage type rape-isatis tinctoria G monomer additional system is applied to the preparation of the medicine for reducing the virus content in the lung of the body after SARS-CoV-2 infection of mice.
In the above applications, preferably, SARS-CoV-2 in mouse SARS-CoV-2 infection is a novel coronavirus SARS-CoV-2/WBP-1, with the collection number being CCTCC NO: v202031.
In the above applications, preferably, the main active ingredient of the drug is an extract of Brassica napus-Isatis indigotica fort monomer addition system, and the extraction method comprises:
mixing cabbage type rape-Isatis tinctoria G monomer addition system powder with methanol, performing ultrasonic treatment at 50-60 deg.C, collecting supernatant, evaporating off methanol to obtain methanol crude extract; dissolving the crude extract with single distilled water, evaporating to remove methanol, extracting with ethyl acetate to obtain lower layer containing effective components, and evaporating water to dry.
The protection scope of the invention also includes: a method for constructing a drug screening model of a mouse infected with a novel coronavirus comprises the following steps: infected with a preservation number of CCTCC NO: v202031 mice, using Brassica napus-Isatis tinctoria G monomer addition line extract as therapeutic drug standard, screening other therapeutic infections with a preservation number of CCTCC NO: v202031.
Compared with the prior art, the invention has the following advantages:
1) the cabbage type rape-isatis tinctoria G monomer addition line extract has obvious SARS-CoV-2 virus resisting effect in cell infection model and no toxic side effect on cell.
2) The cabbage type rape-isatis tinctoria additional line has the advantage of large-scale planting, can provide sufficient raw materials for production and extraction of an anti-SARS-CoV-2 preparation, and has low cost and large yield.
3) The cabbage type rape-isatis tinctoria additional line has the advantages of being natural and healthy, free of toxic and side effects, free of drug resistance and the like.
4) The cabbage type rape-isatis tinctoria additional line can be developed and utilized as antiviral medicinal and edible vegetables, feeds and the like, and can be used as health-care food for human and animals to guarantee the health of the animals and the human.
Drawings
FIG. 1 Effect of different addition lines of Brassica napus-Isatis tinctoria on the proliferation of SARS-CoV-2 virus at the cellular level in vitro.
Wherein A, B, C, D-1, E-1, F-1 and G are rape-isatin monomer addition systems; d-2 and F-2 are binary addition systems.
FIG. 2 is a schematic representation of the cytotoxicity of Brassica napus-Isatis tinctoria G monomer addition line extract on Caco-2 cells.
FIG. 3 the effect of different amounts of Brassica napus-Isatis tinctoria G monomer addition line extract on the proliferation of SARS-CoV-2 virus.
FIG. 4 is a graph showing the effect of Brassica napus-Isatis tinctoria G monomer addition line extract on the body weight of SARS-CoV-2 virus infected mice. FIG. 5 is a schematic representation of the effect of Brassica napus-Isatis tinctoria G monomer addition line extracts on the survival of mice infected with SARS-CoV-2 virus.
FIG. 6 is a schematic diagram showing the effect of Brassica napus-Isatis tinctoria G monomer addition line extract on the lung morphology of SARS-CoV-2 virus infected mice.
FIG. 7 Effect of Brassica napus-Isatis tinctoria G monomer addition line extract on lung histopathological changes of SARS-CoV-2 virus infected mice.
The specific implementation mode is as follows:
for a better understanding of the present disclosure, the following examples are provided to illustrate the present disclosure, but the present disclosure is not limited to the following examples. Unless otherwise specified, the test methods and conditions in the examples of the present invention are conventional methods. The technical scheme of the invention is a conventional scheme in the field if no special description exists; the reagents or materials are commercially available, unless otherwise specified. All tests related to the live SARS-CoV-2 virus were performed in the biosafety third-level laboratory (ABSL 3).
Currently, antiviral drug evaluation models are largely classified into an in vitro model (in vitro model) and an in vivo model (in vivo model).
The in vitro model mainly uses various cell lines to evaluate the medicine, and has the advantages of providing a large number of cells with the same genetic character as research objects, being convenient to operate, eliminating the influence of other external factors, detecting the toxicity, effective concentration and the like of the medicine, and providing more bases for later mechanism research. In vivo models various models of animal infection are generally used, and the overall effect of the drug in living animals is measured by various phenotypic indicators after drug treatment. The method has the advantage of being capable of carrying out real and systematic evaluation on the effect of the candidate drug in the living body. In the invention, the human colon cancer cell line Caco-2 is adopted to measure the in vitro anti-SARS-CoV-2 effect of the cabbage type rape-isatis indigotica fort different addition line extracts; the in vivo anti-SARS-CoV-2 effect of the Brassica napus-Isatis tinctoria addition line extract was evaluated using a mouse adapted strain WBP-1 infection model.
Experimental materials:
(1) cell line, experimental animal and virus required by experiment
Cell line: caco-2 cells were stored in the university of Huazhong agriculture laboratory;
experimental animals: SPF grade 4 to 6 week old Balb/c mice, purchased from the Experimental animals center of the university of Sanxia;
the novel coronavirus SARS-CoV-2/WBP-1 has the preservation number of CCTCC NO: v202031, which has been disclosed in CN111961653A, and in the present invention, the virus is referred to simply as novel coronavirus WBP-1 or mouse adapted strain WBP-1;
(2) cabbage type rape-woad additional line required by experiment and parent thereof
The Brassica napus-Isatis tinctoria addition line and its parent used in the experiment are provided by the team of Li-Byun teachers at plant science and technology institute of Huazhong university
The cabbage type rape-woad addition line is formed by hybridizing somatic cells of cabbage type rape and woad, a formed somatic Cell hybrid and the cabbage type rape are backcrossed continuously for multiple generations, 7 chromosomes of woad are respectively added to the rape, and then 7 monomer addition lines (A, B, C, D, E, F, G) are obtained, wherein a specific preparation process of the cabbage type rape-woad monomer addition line is described in 'Development of a complex of monomeric addition lines between Brassica napus and Isatis indica' (Kang Lei et al,2014, Plant Cell); reference is made to the creation and cytological analysis of the Brassica napus-Isatis indigotica fort binary addition line (Yang Han et al, proceedings of oil crops, 2016, 38(3): 281) 286).
(3) Reagents required for the experiment:
DMEM medium, Fetal Bovine Serum (FBS) were purchased from GIBCO;
cell activity detection kit: TransDetect Cell Counting Kit (CCK) was purchased from Beijing Quanji gold organisms.
Example 1:
preparation of brassica napus-isatis indigotica additional line extract:
harvesting the whole wild cabbage type rape-woad different addition lines planted in the field, naturally drying in the shade or drying at low temperature, crushing, sieving by a 80-mesh sieve, and collecting powder for preparing coarse extraction, wherein the specific extraction steps are as follows:
1) 100g of different attachment system powders and 1000mL of methanol were added to a 2000mL Erlenmeyer flask. Ultrasonic treating at 55 deg.C for 40-60min, filtering, and collecting supernatant and residue respectively;
2) adding 1000mL of methanol into the filter residue in the step 1, continuing ultrasonic treatment, circulating for 5-6 times according to the step 1, and combining all obtained supernatants;
3) evaporating methanol from the supernatant in the step 2 at 50 ℃ in a rotary evaporator;
4) dissolving the crude extract with single distilled water to evaporate methanol, extracting with equal volume of ethyl acetate, collecting the lower layer containing effective components, flowing out from the lower end of separating funnel, collecting the upper layer containing ethyl acetate, recovering the upper layer, and repeatedly extracting the lower layer for 4-5 times until the upper layer becomes clear light color;
5) evaporating water from the extracted lower water-soluble substance at 50 ℃ by using a rotary evaporator, taking down the rotary bottle, placing the rotary bottle in a 50 ℃ oven, drying and weighing;
6) dissolving the dried extract with single distilled water, transferring into a sterilization bottle, and sealing with ethyl acetate solution for later use.
The extraction method of Brassica napus and Isatis tinctoria parents is the same as above.
Example 2:
effect of Brassica napus-Isatis tinctoria different addition line extracts on the proliferation of SARS-CoV-2 Virus at the in vitro cellular level
1. Cell culture
After 2 passages, the frozen and recovered Caco-2 cells are subjected to expanded culture by using a DMEM medium containing 10% fetal calf serum and double antibodies (penicillin 100U/ml and streptomycin 100 ug/ml).
2. Brassica napus-isatis additional line extract for treating cells and infecting with new coronavirus WBP-1
1) Digesting and passaging Caco-2 cells with good growth state, and adjusting cell density to 1 × 10 with cell culture medium5Per ml, 1ml per well was inoculated in 12-well plates.
2) When the cell growth monolayer density reaches 80%, extracts of different addition lines of the cabbage type rape-isatis indigotica fort are added into Caco-2 cells in a 12-well plate, the concentration of the extracts is 200mg/mL, and the working concentration is 400 mu g/mL.
3) Treating different addition line extracts for 24h, infecting with new coronavirus (MOI of 1), incubating at 37 deg.C for 1h, discarding virus incubation solution, adding culture medium containing corresponding addition line extract with the same concentration, and further culturing for 24 h.
4) And collecting virus cell culture solution after 24h, and performing immunoblotting detection.
3. Western blot experiment (Western Blotting)
After Caco-2 cells are infected by the new coronavirus (WBP-1), the influence of the brassica napus-isatis tinctoria additional line extract on the proliferation of the new coronavirus is verified by a Western Blotting experiment, and the specific steps are as follows:
1) and (3) adding 5x protein loading into the cell culture virus solution collected in the step (2), boiling in boiling water for 10min, and centrifuging for later use.
2) SDS-PAGE electrophoresis: preparing 10% polyamide gel as separation gel, performing sample application electrophoresis, keeping constant pressure of concentrated gel at 80V for 30min, and keeping constant pressure of separation gel at 120V for 1-1.5 h.
3) Film transfer: and pre-cooling the membrane transferring liquid in advance during membrane transferring, putting the PAGE gel subjected to electrophoresis into a membrane transferring groove, and transferring the membrane for 1h at constant pressure of 100V.
4) And (3) sealing: and taking out the NC membrane after the membrane is transferred, and sealing the NC membrane for 1h by using sealing liquid.
5) Primary antibody and secondary antibody incubation: the blocked NC membrane was incubated for 2h or overnight at 4 deg.C, and after removal of the primary antibody, washed 6 times with 1 × TBST, 5min each time. The primary antibody was incubated for membrane washing followed by incubation with HRP-labeled secondary antibody for 1h, 6 washes with 1x TBST, 5min each.
6) Color development: color development was performed using Thermo's ECL color development kit.
As shown in FIG. 1, of the 9 addition lines of Brassica napus-Isatis tinctoria, both the F-1 and G-monomer addition lines had some effect against the novel coronavirus, while the other 7 addition lines had no significant difference from the control, i.e., no inhibitory effect on the novel coronavirus WBP-1. The G monomer has obvious effect of resisting new coronavirus, the monomer has stronger inhibition effect on the new coronavirus WBP-1 than isatis tinctoria, the multiplication of the new coronavirus on cells can be obviously inhibited, and after the virus is infected, the virus content in the cell fluid of the G monomer treatment group is obviously lower than that of an untreated infected control group.
Example 3:
inhibition of SARS-CoV-2 virus proliferation by Brassica napus-Isatis tinctoria G monomer addition lines at the in vitro cell level:
according to the results of example 2, the G monomer addition line extract has a significant inhibitory effect on the new coronavirus at the working concentration of 4mg/mL, and example 3 further confirms the anti-new coronavirus effect of the G monomer addition line extract and the inhibitory effect of different using concentrations thereof on the proliferation of the new coronavirus.
1. Toxic effects of monomeric G extract on cells
1) Digesting and passaging Caco-2 cells which grow well, and adjusting the cell density to be 2 multiplied by 10 by using cell growth liquid (DMEM culture medium + 10% fetal calf serum + double antibody)6Inoculating 96-well plates with each well being 100 mul;
2) mu.l of G monomer extract prepared from culture medium (DMEM medium + 10% serum + double antibody) and different concentrations was added to each well and mixed well. Setting 7 concentration gradients, wherein the concentration of the added G monomers is 0 mug/mL respectively; 50 mu g/mL; 100 mu g/mL; 200 mu g/mL; 400 mug/mL; 800 mug/mL; 1600 ug/mL.
3) Setting cell control at the same time, placing at 37 deg.C and 5% CO2Culturing in an incubator.
4) Culturing for 48h, measuring cell activity with TransDetect cell counting box, adding 10 μ l of CCK reagent into 96-well plate cell after culturing, incubating at 37 deg.C in dark for 2h, detecting OD450nm reading with microplate reader, and calculating
Cell survival rate:
cell survival (%). percent vs. drug treated/untreated control 100%
As shown in FIG. 2, the cell viability was measured in response to the toxic effect of the G monomer extract on Caco-2 cells, and it can be seen from FIG. 2 that the cell activity was above 80% at a working concentration of G monomer of 400. mu.g/mL.
2. Inhibitory Effect of monomer extract of G at various concentrations on SARS-CoV-2 Virus proliferation
1) Digesting and passaging Caco-2 cells with good growth state, and adjusting cell density to 1 × 10 with cell culture medium5Per ml, 1ml per well was inoculated in 12-well plates.
2) When the cell growth monolayer density reached 80%, the G monomer extract was added to the 12-well plate at a concentration of 200mg/mL, and the concentrations of the G monomer extract were 0. mu.g/mL, 100. mu.g/mL, 200. mu.g/mL, and 400. mu.g/mL, respectively.
3) Treating the G monomer extracts with different concentrations for 24h, infecting new coronavirus (MOI is 1), incubating in an incubator at 37 ℃ for 1h, discarding virus incubation liquid, adding culture medium containing G monomer extract with corresponding concentration, and continuing to culture cells for 24 h.
4) After 24h, the virus cell culture fluid was collected and subjected to immunoblotting (Western Blotting) detection.
As shown in FIG. 3, the WB assay showed that the intensity of the SARS-CoV-2-NP protein decreased with the increase of the concentration of the G monomer, indicating that the extraction of the G monomer had an inhibitory effect on the proliferation of SARS-CoV-2 and was dose-dependent, and that the inhibitory effect on SARS-CoV-2 increased with the increase of the amount of the G monomer extract.
Example 4:
evaluation of antiviral Effect of G monomer extract in mouse lethal infection model
The in vivo effect evaluation of the G monomer against the new coronavirus is carried out in a lethal infection model of the new coronavirus mouse, and the mouse animal test comprises the following specific steps:
1) randomly dividing female BALB/c mice with the age of 6 weeks into 2 major groups, wherein the first major group is a group without virus infection, and is respectively a PBS control group, a Brassica napus (Brassica napus) parent group, an Isatis indigotica (Isatis indigotica) parent group and a G monomer addition line group, and each group comprises 8 mice; the second major group is a group infected with SARS-CoV-2 virus, which is PBS + SARS-CoV-2 control group, Brassica napus (Brassica napus) parent + SARS-CoV-2 group, Isatis indigotica (Isatis indigotica) parent + SARS-CoV-2 group, and G monomer addition line group + SARS-CoV-2, each group contains 10 mice.
2) Gavage was performed on mice with Brassica napus, Isatis tinctoria, and G monomer 3 plant extract and PBS for one week, once a day, and 100. mu.L (200mg/mL) per mouse.
3) One week after the plant extract is infused, new coronavirus WBP-1 challenge infection is carried out, and each mouse is infected with 2 × 10 drops of nose5PFU novel coronavirus mouse adapted strain SARS-CoV-2/WBP-1, infection dose 50. mu.l, mice body weight and survival were observed and recorded daily for 10 consecutive days after virus infection.
4) And drawing a weight change curve and a survival rate curve according to the statistical result.
5) 40 female BALB/c mice of 6 weeks old were randomly divided into 4 groups, which were PBS + SARS-CoV-2 control group, Brassica napus (Brassica napus) parent + SARS-CoV-2 group, Isatis indigotica (Isatis indigotica) parent + SARS-CoV-2 group, and G monomer addition line group + SARS-CoV-2, 10 mice per group, and the administration and infection were performed as above, and the mice were dissected on day 5 after SARS-CoV-2 infection, and 3 mice per group were dissected randomly, and lung tissue samples were taken, and the collected lung tissue samples were fixed with tissue fixative (4% PFA) for histopathological analysis.
The results of anti-SARS-CoV-2 in mice show that the G monomer also has better anti-new coronavirus effect in animals.
The weight change results of the mice are shown in fig. 4 and table 1, the mice which are not infected with the new coronavirus and are drenched with the PBS and the 3 plant extracts have no weight reduction during the monitoring period, and the weight gradually increases along with the increase of the daily age, which indicates that the G monomer and the parent extracts thereof have no influence on the growth and the health of the mice, and the use is healthy and safe; in all 4 groups infected with the new coronavirus, the body weight of mice began to decrease after challenge, the body weight of the PBS group and the isatis indigotica parent group gradually decreased after infection and all died on day 7 after infection, the body weight of the rape group also continuously decreased after infection and all died on day 8 after infection, but the body weight of the G monomer group began to recover on day 9 after decrease.
TABLE 1 Effect of Brassica napus-Isatis tinctoria G monomer addition line extracts on the body weight of mice infected with SARS-CoV-2 Virus
The results of the survival rate of mice are shown in fig. 5 and table 2, and the mice were all alive in 4 groups not infected with the new coronavirus (in fig. 5, the lines of the PBS group, the Brassica napus group, the Radix lsatidis group, and the G monosome group coincide); survival of mice in G monomer treated group was 50% in 4 groups infected with the corresponding new coronavirus; the survival rate of mice in the rape parent group, the isatis indigotica parent group and the PBS group is 0.
TABLE 2 Effect of Brassica napus-Isatis tinctoria G monomer addition line extracts on the survival of mice infected with SARS-CoV-2 Virus
The appearance morphology of lung tissues shows that the G monomer extract can relieve the change of the morphology of lung tissues caused by the infection of new coronavirus (figure 6); the lung histopathological section detection result is shown in fig. 7: severe inflammatory cell infiltration, pulmonary congestion, was observed in the lungs of the control mice, whereas the damage of the lung tissue by the virus in the G monomer addition line group was significantly reduced.
The above results indicate that the G monomer addition line has significant anti-new coronavirus effects both at the cellular level and in mice. In view of the above results, the G monomer can be used for the preparation of a preparation for preventing or treating SARS-CoV-2 infection.
Claims (5)
1. Application of Brassica napus-Isatis tinctoria G monomer addition system in preparation of preparation for treating or preventing novel coronavirus SARS-CoV-2 infection is provided.
2. The application of cabbage type rape-isatis tinctoria G monomer addition system in the preparation of SARS-CoV-2 inhibiting medicine;
the application of cabbage type rape-isatis tinctoria G monomer additional system in preparing medicine for preventing and treating SARS-CoV-2 infection;
the application of cabbage type rape-isatis tinctoria G monomer addition system in the preparation of medicines for relieving the weight reduction after SARS-CoV-2 infection;
the application of cabbage type rape-isatis tinctoria G monomer addition system in preparing medicine for raising the survival rate of SARS-CoV-2 infected body;
the application of cabbage type rape-isatis tinctoria G monomer additive in preparing medicine for lowering virus content in body's lung after SARS-CoV-2 infection.
3. The use according to claim 3 or 4 or 5 or 6, wherein the infected subject is a mouse, and the collection number of the infected SARS-CoV-2 virus is CCTCC NO: v202031.
4. The use according to claim 1 or 2 or 3 or 4 or 5 or 6, wherein the main effective component of the medicament is Brassica napus-Isatis indigotica fort monomer addition line extract, and the extraction method comprises:
mixing cabbage type rape-Isatis tinctoria G monomer addition system powder with methanol, performing ultrasonic treatment at 50-60 deg.C, collecting supernatant, evaporating off methanol to obtain methanol crude extract; dissolving the crude extract with single distilled water, evaporating to remove methanol, extracting with ethyl acetate to obtain lower layer containing effective components, and evaporating water to dry.
5. A method for constructing a drug screening model of a mouse infected with a novel coronavirus comprises the following steps: infected with a preservation number of CCTCC NO: v202031 mice, using Brassica napus-Isatis tinctoria G monomer addition line extract as therapeutic drug standard, screening other therapeutic infections with a preservation number of CCTCC NO: v202031.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110478468.1A CN113181229B (en) | 2021-04-30 | 2021-04-30 | Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110478468.1A CN113181229B (en) | 2021-04-30 | 2021-04-30 | Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113181229A true CN113181229A (en) | 2021-07-30 |
| CN113181229B CN113181229B (en) | 2021-10-29 |
Family
ID=76982957
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110478468.1A Active CN113181229B (en) | 2021-04-30 | 2021-04-30 | Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113181229B (en) |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105230486A (en) * | 2015-09-29 | 2016-01-13 | 华中农业大学 | Breeding method of cabbage type rape woad oil cytoplasm male sterile line |
-
2021
- 2021-04-30 CN CN202110478468.1A patent/CN113181229B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105230486A (en) * | 2015-09-29 | 2016-01-13 | 华中农业大学 | Breeding method of cabbage type rape woad oil cytoplasm male sterile line |
Non-Patent Citations (5)
| Title |
|---|
| LEI KANG等: "Development of a complete set of monosomic alien addition lines", 《PLANT CELL REP》 * |
| 康雷等: "利用菘蓝创建抗病毒油菜及新的雄性不育/恢复系统 ", 《中国油料作物学报》 * |
| 杜雪竹等: "甘蓝型油菜与菘蓝原生质体融合及植株再生体系的研究 ", 《湖北农业科学》 * |
| 杨健等: "蓝菜1号特征特性及标准化栽培技术 ", 《中国现代中药》 * |
| 杨汉等: "甘蓝型油菜―菘蓝二体附加系的创制和细胞学分析 ", 《中国油料作物学报》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113181229B (en) | 2021-10-29 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102274314A (en) | Evergreen coccidian powder and preparation method thereof | |
| CN102824417B (en) | New method for treating helicobacter pylori related diseases | |
| CN101596246B (en) | Smoked plum extractive and wild jujube seed extractive compound preparation as well as preparation method and application thereof | |
| CN108926584A (en) | The antimicrobial purposes of chimonanthea extract | |
| CN1330330C (en) | Antivirus medicinal composition, preparation method and use | |
| CN107137434B (en) | Pharmaceutical composition, preparation method thereof and application thereof in preparation of coxsackie virus resistant medicine | |
| CN113181229B (en) | Application of cabbage type rape-isatis tinctoria G monomer addition system in inhibiting novel coronavirus SARS-CoV-2 | |
| CN115105502B (en) | Application of compound containing stephania plant alkaloid in preparation of cat infectious peritonitis medicine | |
| CN108635391A (en) | A kind of hempleaf groundsel herb phenolic acid components and the preparation method and application thereof | |
| CN113318141A (en) | Application of agrimony extract | |
| CN112957388B (en) | Application of brassica napus-isatis tinctoria E monomer addition system in inhibiting influenza virus | |
| CN106084084B (en) | The preparation method and its usage of the raw honeysuckle total polysaccharide extractive of rock | |
| CN108926585A (en) | Application of the wintersweet genus plants extract in pharmacy | |
| CN108926583A (en) | The purposes of Zhejiang wintersweet anti-microbial infection | |
| CN107582782A (en) | A kind of process for preparing medicine of composition containing betel nut and its anti anoxia application | |
| CN103655757A (en) | Traditional Chinese medicine compound for effectively resisting swine influenza virus | |
| CN112891343B (en) | Application of 3-indole acetonitrile in preparing medicine for inhibiting novel coronavirus SARS-CoV-2 | |
| CN104523809B (en) | A kind of preparation method of Fructus Corni pigment | |
| CN102697800A (en) | Chickweed polysaccharide composition and applications thereof in preparation of antiviral medicine | |
| CN102078529B (en) | Chinese medicinal preparation for treating common cold in children | |
| CN107080761B (en) | The purposes of the anti-Pseudorabies virus of wintersweet platymiscium | |
| CN108992478A (en) | The application of Aralia wood extract in medicine preparation | |
| CN107158052A (en) | Application of the Herba Melastomatis dodecandri extract in medicine is prepared | |
| CN104983777A (en) | Preparation method for radix astragali and licorice powder and product thereof | |
| CN104758340A (en) | Caffeoylquinic acid extract in Lonicera japonica Thunb leaf, preparation method and application thereof |
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 |