CN103191099B - The application in anti-listeria infection medicine prepared by fisetin - Google Patents
The application in anti-listeria infection medicine prepared by fisetin Download PDFInfo
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- CN103191099B CN103191099B CN201310138824.0A CN201310138824A CN103191099B CN 103191099 B CN103191099 B CN 103191099B CN 201310138824 A CN201310138824 A CN 201310138824A CN 103191099 B CN103191099 B CN 103191099B
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- Prior art keywords
- fisetin
- listeria
- listerella
- mice
- infection
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- XHEFDIBZLJXQHF-UHFFFAOYSA-N fisetin Chemical compound C=1C(O)=CC=C(C(C=2O)=O)C=1OC=2C1=CC=C(O)C(O)=C1 XHEFDIBZLJXQHF-UHFFFAOYSA-N 0.000 title claims abstract description 46
- 235000011990 fisetin Nutrition 0.000 title claims abstract description 23
- 206010024641 Listeriosis Diseases 0.000 title claims abstract description 10
- 239000003814 drug Substances 0.000 title claims abstract description 9
- 241000186781 Listeria Species 0.000 claims abstract description 18
- 208000015181 infectious disease Diseases 0.000 claims abstract description 7
- 241000186779 Listeria monocytogenes Species 0.000 claims description 3
- 230000001580 bacterial effect Effects 0.000 claims description 3
- 208000035143 Bacterial infection Diseases 0.000 claims 2
- 208000022362 bacterial infectious disease Diseases 0.000 claims 2
- 239000003937 drug carrier Substances 0.000 claims 1
- 241000699670 Mus sp. Species 0.000 abstract description 12
- 210000004027 cell Anatomy 0.000 abstract description 9
- 241000699666 Mus <mouse, genus> Species 0.000 abstract description 6
- 210000003024 peritoneal macrophage Anatomy 0.000 abstract description 5
- 230000002949 hemolytic effect Effects 0.000 abstract description 4
- 230000007306 turnover Effects 0.000 abstract description 4
- 241001494479 Pecora Species 0.000 abstract description 3
- 230000003115 biocidal effect Effects 0.000 abstract description 3
- 230000000694 effects Effects 0.000 abstract description 3
- 210000003743 erythrocyte Anatomy 0.000 abstract description 3
- 206010059866 Drug resistance Diseases 0.000 abstract description 2
- 230000002401 inhibitory effect Effects 0.000 abstract description 2
- 238000009509 drug development Methods 0.000 abstract 1
- 239000003596 drug target Substances 0.000 abstract 1
- 239000002547 new drug Substances 0.000 abstract 1
- 238000002560 therapeutic procedure Methods 0.000 abstract 1
- 206010018910 Haemolysis Diseases 0.000 description 5
- 230000008588 hemolysis Effects 0.000 description 5
- 108010006464 Hemolysin Proteins Proteins 0.000 description 4
- 239000003228 hemolysin Substances 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 3
- 238000011725 BALB/c mouse Methods 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 2
- MYSWGUAQZAJSOK-UHFFFAOYSA-N ciprofloxacin Chemical compound C12=CC(N3CCNCC3)=C(F)C=C2C(=O)C(C(=O)O)=CN1C1CC1 MYSWGUAQZAJSOK-UHFFFAOYSA-N 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 230000001225 therapeutic effect Effects 0.000 description 2
- 206010000234 Abortion spontaneous Diseases 0.000 description 1
- 241000208223 Anacardiaceae Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 241000751185 Listeria monocytogenes ATCC 19115 Species 0.000 description 1
- 241000092431 Listeria monocytogenes EGD Species 0.000 description 1
- 201000009906 Meningitis Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010027906 Monocytosis Diseases 0.000 description 1
- 206010040047 Sepsis Diseases 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- 235000009392 Vitis Nutrition 0.000 description 1
- 241000219095 Vitis Species 0.000 description 1
- 208000007502 anemia Diseases 0.000 description 1
- 239000003242 anti bacterial agent Substances 0.000 description 1
- 230000003110 anti-inflammatory effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 229940088710 antibiotic agent Drugs 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229960003405 ciprofloxacin Drugs 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 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
- 229940079593 drug Drugs 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 244000078673 foodborn pathogen Species 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000002458 infectious effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 150000002555 kaempferol Chemical class 0.000 description 1
- 230000031700 light absorption Effects 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 208000015994 miscarriage Diseases 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 208000013223 septicemia Diseases 0.000 description 1
- 208000000995 spontaneous abortion Diseases 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 239000002023 wood Substances 0.000 description 1
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- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
The present invention relates to fisetin and preparing the application in anti-listeria infection medicine, by sheep red blood cell hemolytic test, mice huge bite like cell J774 and Turnover of Mouse Peritoneal Macrophages damage protection test and mice Listerella infection model confirm its anti-listeria infection effect.With compared with antibiotic therapy, use fisetin treatment to have and have no drug resistance, the feature that cure rate is high, therefore fisetin may be used for new drug development, and confirms important in inhibiting to drug targets.
Description
Technical field
The present invention relates to fisetin preparing the application in anti-listeria infection medicine, belonging to field of medicaments.
Background technology
Fisetin (Fisetin) is the natural kaempferol compounds of isolated one from the plants such as Anacardiaceae plant wood La Shu, in the vegetables such as Fructus Fragariae Ananssae, Fructus Vitis viniferae, Fructus Mali pumilae, Bulbus Allii Cepae and fruit, also have existence.There is obvious antioxidation, anti-inflammatory property, certain regulating action is also had to immune system, but have no the report that fisetin infects for anti-listeria so far both at home and abroad.
Listeria monocytogenes (Listeria monocytogenges, LM) is called for short Listeria monocytogenes, is a kind of important Amphixenosis pathogen, is also foodborne pathogens important on the most common in food, public hygienics.This bacterium can infect people, multiple domestic animal and birds, causes meningitis, septicemia, anemia of pregnant woman's miscarriage and monocytosis, neonate, old people, pregnant woman and immunologic hypofunction person more easy infection morbidity.Though this microbial disease incidence is low, case fatality rate can up to 30% ~ 70%.In recent years, antibiotic a large amount of use causes Bacterial Drug Resistance of Patients to increase, and has been found that various clinical separated strain is serious to the anti-biotic resistance such as tetracycline and ciprofloxacin, causes the treatment of listeriosis to face the situation pasted medical help.Therefore, novel, safe medicine is found extremely urgent.Fisetin comes from China's Chinese medicine, and the present invention's research confirms that fisetin has good therapeutic effect to anti-listeria infection.
Summary of the invention
The molecular structure of fisetin is as follows:
The present invention by sheep red blood cell hemolytic test, mice huge bite like cell J774 and Turnover of Mouse Peritoneal Macrophages damage protection test and mice Listerella infection model confirm its anti-listeria infection effect.
Detailed description of the invention
1. hemolytic test
The Listerella hemolysin of purification does continuous print doubling dilution in 96 orifice plates, adds 10 in each hole
7individual sheep red blood cell, centrifugal hatch 30h in 37 DEG C after.Haemoclastic most highly diluted multiple is made to be considered as haemolysis titre (Hemolysis titre).
Do not add fisetin and add variable concentrations fisetin after the haemolysis titre of Listerella hemolysin as following table 1:
Table 1. fisetin is to the inhibitory action of Listerella hemolysin hemolytic activity
| Fisetin (μ g/ml) | Haemolysis titre (Hemolysis titre) |
| 0 | 2048 |
| 16 | 16 |
| 8 | 64 |
| 4 | 256 |
| 2 | 512 |
2. the huge protection test of biting like cell J774 and Turnover of Mouse Peritoneal Macrophages damage of mice
Listerella L.monocytogenes EGD and ATCC 19115 is cultured to OD600nm=0.8 in TSB culture medium, gets the bacterial cultures of 5ml, centrifugal and be resuspended in the DMEM culture medium of 10ml.J774 cell in DMEM culture medium (adding 10% hyclone) in 37 DEG C, 5%CO
2cultivate after 24 hours, with 2 × 10
4individual cells/well is laid in 96 porocyte culture plates, every hole 200 μ l.After cell attachment, add 100 μ l Listeria suspensions, and add the fisetin of variable concentrations, be placed in cell culture incubator in 37 DEG C, 5%CO
2dual culture 5h.Centrifuging and taking 100 μ l supernatant, and add LDH reagent 100 μ l, hatch 30 minutes for 37 DEG C, rear microplate reader surveys 570nm light absorption value, compares the cell LDH release rate of variable concentrations fisetin process.Be separated peritoneal macrophage from BALB/c mouse, be then laid in 96 porocyte culture plates with same cell quantity, subsequent operation is the same.
Result shows, fisetin can protect that the mice of Listerella hemolysin mediation is huge bites like cell J774 and Turnover of Mouse Peritoneal Macrophages damage, and this effect presents dose dependent.
3. the experimental therapeutic that mice Listerella is infected is studied
3.1 mice Listerella infection models
BALB/c mouse (male, 18-22g), through tail vein injection 50 μ l Listerella suspension (Listerella ATCC 19115), sets up mice Listeria infection model.
3.2 protective rate tests
The fisetin of 2h difference subcutaneous injection 100,50 and 25mg/kg (100 μ l) behind mouse inoculation Listerella, every 6h is administered once.Not administration matched group gives the normal saline with 100 μ l, often organizes 50 mices.After Dosage Regimens Dosage, record mouse infection Listerella 24h respectively, the mortality rate after 48h, 72h.
Result shows, after fisetin drug treatment, significantly reduces the mortality rate (P < 0.05) infected mice Listerella.As table 2.
Table 2. fisetin is on the impact of mice Listerella infectious age
Claims (4)
1. the application in anti-listeria infection medicine prepared by fisetin.
2. apply as claimed in claim 1, it is characterized in that described Listerella is infected and refer to by the bacterial bacterial infection of listeria.
3. apply as claimed in claim 2, wherein bacterial infection refers to the infection caused by Listeria monocytogenes.
4. apply as claimed in claim 1, it is characterized in that described pharmaceutical pack contains pharmaceutically acceptable carrier.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310138824.0A CN103191099B (en) | 2013-04-22 | 2013-04-22 | The application in anti-listeria infection medicine prepared by fisetin |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310138824.0A CN103191099B (en) | 2013-04-22 | 2013-04-22 | The application in anti-listeria infection medicine prepared by fisetin |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103191099A CN103191099A (en) | 2013-07-10 |
| CN103191099B true CN103191099B (en) | 2015-10-07 |
Family
ID=48714038
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310138824.0A Active CN103191099B (en) | 2013-04-22 | 2013-04-22 | The application in anti-listeria infection medicine prepared by fisetin |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103191099B (en) |
-
2013
- 2013-04-22 CN CN201310138824.0A patent/CN103191099B/en active Active
Non-Patent Citations (1)
| Title |
|---|
| Influence of phenolic compounds from wines on the growth of Listeria monocytogenes;María J. Rodríguez Vaquero等;《Food Control》;20071231;第18卷;第587–593页 * |
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| Publication number | Publication date |
|---|---|
| CN103191099A (en) | 2013-07-10 |
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| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
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Effective date of registration: 20191127 Address after: 262400 Nine-level Village South of Zhuliu Street, Changle County, Weifang City, Shandong Province Co-patentee after: SHANDONG JINZHUJI PHARMACEUTICALS Co.,Ltd. Patentee after: SHANDONG GUANGYUAN PHARMACEUTICAL TECHNOLOGY Co.,Ltd. Address before: 130012 Changchun Qianjin Street, Jilin, No. 2699 Patentee before: Jilin University |
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| PE01 | Entry into force of the registration of the contract for pledge of patent right | ||
| PE01 | Entry into force of the registration of the contract for pledge of patent right |
Denomination of invention: The application of lutein in the preparation of anti Listeria infection drugs Effective date of registration: 20231228 Granted publication date: 20151007 Pledgee: Prudential Bank Ji'nan branch of Limited by Share Ltd. Pledgor: SHANDONG JINZHUJI PHARMACEUTICALS Co.,Ltd.|SHANDONG GUANGYUAN PHARMACEUTICAL TECHNOLOGY Co.,Ltd. Registration number: Y2023980075034 |