CN107648248A - Lamp-dish flower acetic is preparing the application in treating NLRP3 relevant disease medicines - Google Patents
Lamp-dish flower acetic is preparing the application in treating NLRP3 relevant disease medicines Download PDFInfo
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Abstract
The invention discloses lamp-dish flower acetic to prepare the application in treating NLRP3 relevant disease medicines.Find that lamp-dish flower acetic as NLRP3 inflammation corpusculum inhibitor, can suppress the activation of NLRP3 inflammation corpusculum and cell Jiao dies in the present invention.Lamp-dish flower acetic can significantly inhibit the proinflammatory β of inflammation factor IL 1 and danger signal the molecule HMGB1 of LPS+ATP inductions release, suppress cell Jiao to die, and death caused by bacterial sepsis can be effectively reduced, available for preparation preventing and treating, the NLRP3 inflammation corpusculum because of caused by infection or endogenous DAMP activates the medicine that relevant disease is died with cell Jiao.Present invention finds the new activity of lamp-dish flower acetic, it is used to prevent and treat autoimmunity disease caused by acquired diseases associated with inflammation and heredity NLRP3 mutation for exploitation, the medicine of heredity Cryopyrin associated period heat pyrexia syndromes, infective inflammation disease and nerve degenerative diseases provides theoretical foundation.
Description
Technical field
The invention belongs to biomedical and drug field, more particularly to lamp-dish flower acetic is preparing treatment NLRP3 correlation diseases
Application in medicine.
Background technology
Lamp-dish flower acetic (also known as scutellarin) (scutellarin) is the Huang of the separation and Extraction from the Chinese herbal medicines such as fleabane flower
Letones (《Pharmacopoeia of People's Republic of China》2010 editions), molecular formula C21H18O12, its chemical constitution is as follows.Fleabane flower
B prime is yellow needles, is dissolved in dimethyl sulfoxide (DMSO) and ethanol, is insoluble in water.Clinic is mainly used in treating ischemic heart and brain blood
The treatment paralysed after pipe disease, such as angina pectoris, myocardial infarction, apoplexy, cerebral thrombus and cerebrovascular disease, and treated with obvious
Effect.In addition, there is studies have shown that lamp-dish flower acetic that there is obvious antioxidation, can effective scavenging activated oxygen generation, from
And play the protective effect to cardiovascular and cerebrovascular.
NLRP3 is one of danger signal receptor in inherent immunity cell (including macrophage), can be by a variety of cause of diseases
Body associated molecular pattern (PAMP) or damage associated molecular pattern (DAMP) are activated, and form multi-subunit inflammation corpusculum, simultaneously
It is dead to induce being inflamed property of cell --- cell Jiao dies (pyroptosis) so as to trigger strong inflammatory reaction, is that body is consolidated
There is the important component of immune defense system.
Although NLRP3 inflammation corpusculums are the important components of inherent immunity, its abnormal activation and functional disturbance with it is more
The pathologic process of autoimmunity disease is closely related caused by the acquired diseases associated with inflammation of kind and heredity NLRP3 mutation.A variety of generations
Disease and immunologic derangement are thanked, such as fat (obesity), diabetes B (type 2diabetes, T2D), atherosclerosis
(atherosclerosis), gout (gout) etc., it is all relevant with the NLRP3 abnormal activations that endogenous DAMP induces.Fat small
In mouse and human adipose and hepatic tissue, the expression of NLRP3 inflammation corpusculum components and the rise of caspase-1 activity and obese individuals
T2D symptom is directly related；Amylin (IAPP) may be related to the NLRP3 abnormal activations in T2D.Cholesterol is brilliant
Physical efficiency cause the destruction of macrophage lysosome and activate NLRP3 inflammation corpusculum and interleukin 1 (IL-1) family's cell because
The secretion of son, is the major reason for causing atherosclerosis.Endogenous sodium urate crystals (MSU) can also cause in macrophage
NLRP3 is activated, and causes the inflammatory cytokines such as interleukin-1 ' beta ' (IL-1 β) to discharge and inflammatory reaction, this and gout
(gout) there is very strong association.Heredity Cryopyrin associated period heat pyrexia syndromes (CAPS) and NLRP3 work(
Energy gain mutation is relevant, and this kind of mutation activates NLRP3 compositions, causes caspase-1 continuous activations and IL-1 β and IL-
18 excessive secretion.In addition, the generation of nerve degenerative disease and progress and brain damage are also relevant with NLRP3：It is such as multiple
Property sclerosis (multiple sclerosis), Alzheimer disease (Alzheimer ' s disease), Parkinson's disease
(Parkinson ' s disease) etc., NLRP3 activation may be induced with endogenous DAMP (such as misfolded protein) and is produced
The inflammatory cytokines such as IL-1 β are related；Nearest studies have shown that ischemic brain damage is also relevant with NLRP3 activation.Value
It is noted that the occurrence and development of pyemia (sepsis) also with the abnormal activation of inflammation corpusculum (including NLRP3) and excessive
Cell Jiao die correlation.Bacterium infection not only produces a large amount of PAMP in vivo, and bacterium can discharge ATP in itself, can also induce
Monocytes/Macrophages discharge ATP, so as to activate the activation of the inflammation such as NLRP3 corpusculum, cause acute stage strong inflammatory reaction, cause
Excessive Jiao of inherent immunity cell (including macrophage) is died, and inhibition immunocyte substantially increases, and the final body that suppresses is exempted from
Epidemic disease function, patient is set to enter immunosuppressive condition in the pyemia later stage；Meanwhile Jiao of parenchyma dies is also likely to be to draw in organ
Play the major reason of organ dysfunction.Therefore, the generation that the activation of suppression NLRP3 inflammation corpusculums is died with cell Jiao, will be favourable
In the above-mentioned diseases associated with inflammation of alleviation and provide new therapy approach.
The content of the invention
The shortcomings that it is an object of the invention to overcome prior art and deficiency, there is provided lamp-dish flower acetic is preparing treatment
Application in NLRP3 relevant disease medicines, inhibitor of the lamp-dish flower acetic as NLRP3 inflammation corpusculums, can effectively suppress
The activation of NLRP3 inflammation corpusculums and cell Jiao die, available for the medicine for preparing treatment NLRP3 relevant diseases.
The purpose of the present invention is achieved through the following technical solutions：Lamp-dish flower acetic is preparing treatment NLRP3 relevant disease medicines
Application in thing.
Described NLRP3 relevant diseases include itself exempting from caused by acquired diseases associated with inflammation and heredity NLRP3 mutation
Epidemic disease, heredity Cryopyrin associated period heat pyrexia syndromes (cryopyrin-associated periodic
Syndromes), infective inflammation disease and nerve degenerative diseases.
Autoimmunity disease caused by described acquired diseases associated with inflammation and heredity NLRP3 mutation is metabolic disease and exempted from
Epidemic disease is disorderly, including fat (obesity), diabetes B (T2D), atherosclerosis (atherosclerosis) and gout
(gout)。
Described infective inflammation disease includes pyemia (sepsis), endotoxemia (endotoxemia) and courage
Juice deposits (cholestasis).
Described nerve degenerative diseases include multiple sclerosis (multiple sclerosis), Alzheimer disease
(Alzheimer ' s disease) and Parkinson's disease (Parkinson ' s disease).
Lamp-dish flower acetic is preparing the corpusculum activation of suppression NLRP3 inflammation, suppresses cell Jiao and dies, suppresses caspase-1 activation,
Suppress IL-1 β (proinflammatory inflammation factor IL-1 β) secretions, suppress HMGB1 (high mobility group protein B 1) secretions, suppress ASC patches
(ASC speck) formation, suppress ASC oligomer and formed, anti-systemic bacterial infection, and/or in bacteria resistance medication for treating pyemia
Application.
Described systemic bacterial infection is preferably abdominal cavity bacterial infection, and lamp-dish flower acetic is by suppressing LPS sensitization, ATP
The activation of NLRP3 inflammation corpusculum is died with cell Jiao in the macrophage of induction, and then suppresses caspase-1 activation and maturation
The release of the inflammatory factors such as IL-1 β and HMGB1, it is dead caused by abdominal cavity bacterial infection so as to reduce.
Described bacterium is gram negative pathogenic bacteria；Preferably Escherichia coli；More preferably bacillus coli DH 5 alpha.
The medicine of described treatment NLRP3 relevant diseases suppresses the medicines such as NLRP3 inflammation corpusculum activation comprising pharmaceutically
Acceptable auxiliary material and other rise compatibility synergy active ingredient, can be various formulations, as tablet, granule, capsule,
Dripping pill, sustained release agent, oral liquid, injection etc..
After bone marrow derived macrophage (BMDM) is by LPS sensitization (priming) in the present invention, then through secondary signals such as ATP
Stimulate, activate NLRP3 inflammation corpusculums, cause caspase-1 to activate, promote ripe IL-1 β, HMGB1 secretion, ASC patches
(ASC speck) and ASC oligomer formation；The BMDM cells of lamp-dish flower acetic pretreatment LPS activation, can be significantly inhibited
Caspase-1 is activated, and suppresses ripe IL-1 β, HMGB1 secretion and the formation of ASC speck and ASC oligomer.Mouse passes through
After lamp-dish flower acetic gavage, intraperitoneal injection bacterial death living can obviously reduce.Therefore, lamp-dish flower acetic can pass through suppression
Inflammation corpusculum activates and IL-1 β secretion, so as to play the effect of the diseases such as anti-Systemic inflammation and pyemia.
The present invention is had the following advantages relative to prior art and effect：
1st, lamp-dish flower acetic is NLRP3 inflammation corpusculum inhibitor in the present invention, can significantly inhibit LPS sensitization, ATP is induced
The activation of NLRP3 inflammation corpusculum is died with cell Jiao in macrophage, the final activation for suppressing caspase-1 and ripe IL-1 β
With the release of the inflammatory factor such as HMGB1, effectively reduce dead caused by the bacterial sepsises such as abdominal cavity bacterial infection.Fleabane flower second
The above-mentioned new activity and purposes of element, there is not yet open source literature is reported.
2nd, lamp-dish flower acetic is died by suppressing the activation of NLRP3 inflammation corpusculum and cell Jiao in the present invention, and then is suppressed proinflammatory
Inflammation factor IL-1 β and danger signal molecule HMGB1 release.Have to clinical treatment NLRP3 abnormal activation relevant diseases potential
Therapeutic action, available for related drugs develop.
3rd, find that lamp-dish flower acetic has the work that can suppress that NLRP3 inflammation corpusculum activates and cell Jiao dies in the present invention first
With：Lamp-dish flower acetic can significantly inhibit proinflammatory inflammation factor IL-1 β and danger signal the molecule HMGB1 of LPS+ATP inductions release,
Suppress cell Jiao to die and can effectively reduce dead mouse caused by bacterium infection.Clinically there is no at present can effectively prevent bacterium sense
The specific drug that dye or the NLRP3 inflammation corpusculum activation induced by endogenous DAMP and cell Jiao die.Lamp-dish flower acetic conduct
The inhibitor of NLRP3 inflammation corpusculums, activated for preparing preventing and treating NLRP3 inflammation corpusculum because of caused by infection or endogenous DAMP
The application of medicine is died with cell Jiao, to clinical treatment NLRP3 relevant disease (such as gouts；Heredity Cryopyrin associated periods
Fever syndrome；Neurogenic disease and brain damage：Such as multiple sclerosis, Alzheimer disease, Parkinson's disease；It is infectious
Diseases associated with inflammation, such as pyemia.) etc. it is significant, available for related drugs develop.
Brief description of the drawings
Fig. 1 is the mouse bone marrow cells source property macrophage (BMDM) that lamp-dish flower acetic (scutellarin) is induced LPS+ATP
The immunoblotting assay figure of the inhibitory action of inflammation corpusculum activation and immunodetection (CBA) the analysis culture supernatant based on microballon
Middle IL-1 β level view；Wherein, it is immunoblotting assay figure to scheme A；Scheme the level view that B is IL-1 β in culture supernatant.
Fig. 2 is the mouse bone marrow cells source property macrophage (BMDM) that lamp-dish flower acetic (scutellarin) is induced LPS+ATP
Immunofluorescence microscopy analysis ASC and NLRP3 distribution and formation and the immunoblotting assay of ASC patches (ASCspeck)
The formation figure of ASC oligomer；Wherein, the immunofluorescence microscopic analysis figure that A is ASC and NLRP3 is schemed, figure B is to contain ASC patches
Cell percentages analyze histogram, and figure C is the immunoblotting assay figure that ASC oligomer is formed.
Fig. 3 is the mouse bone marrow cells source property macrophage (BMDM) that lamp-dish flower acetic (scutellarin) is induced LPS+ATP
Propidium iodide (PI) the fluorescent staining figure and immunoblotting assay figure for the inhibitory action that cell Jiao dies；Wherein, it is propidium iodide to scheme A
(PI) fluorescent staining micrograph, figure B are the percentage analysis figure that Jiao dies cells on total cells, and figure C is immunoblotting assay figure.
Fig. 4 is the analysis chart for the death rate that lamp-dish flower acetic (scutellarin) reduces abdominal cavity bacterial infection mouse.
Embodiment
With reference to embodiment, the present invention is described in further detail, but the implementation of the present invention is not limited to this.
Used material in embodiment：
Lamp-dish flower acetic (Scutellarin) is purchased from Guangdong Provincial Medicines Checkout station.Lipopolysaccharides (LPS), atriphos
(ATP), double (the N- hydroxyls of propidium iodide (PI), Hoechst 33342, dimethyl sulfoxide (DMSO) (DMSO), NaTDC, suberic acid
Succinimide ester) (disuccinimidyl suberate) be purchased from Sigma-Aldrich companies.Cell culture complete medium
DMEM, penicillin/streptomycin, hyclone (FBS) are purchased from ThermoFisher companies.Anti- caspase-1 antibody is purchased from Santa
Cruz companies.Anti- NLRP3 antibody is purchased from Adipogen AG companies.Anti-il-i-beta, HMGB1, ASC, 'beta '-tubulin (β-
Tubulin antibody is Cell Signaling Technology Products.Cytometric Bead Array(CBA)
Mouse IL-1 β Flex Set kits are purchased from BD companies.Trichloroacetic acid (TCA) is purchased from Guangzhou Chemical Reagent Factory.Cell in vitro
Experiment lamp-dish flower acetic is dissolved in DMSO, and experiment in vivo lamp-dish flower acetic is dissolved in the PBS solution containing 2% (v/v) Tween-80.
Embodiment 1
(1) cell culture and processing：
L929 cells are purchased from Kunming Institute of Zoology, Chinese Academy of Sciences's cell bank.Cell culture processes are tested with reference to Monack
The operation sequence (Stanford University) of room, by 1 × 108Individual cell culture is 500cm in floor space2Triangular flask
In, culture medium is DMEM complete mediums (200mL), and collection supernatant freezes standby after cultivating 14 days.
The cultural method of mouse bone marrow cells source property macrophage (BMDM)：C57BL/6 mouse, 6-8 weeks, female, purchased from south
Medical university's Experimental Animal Center.After cervical dislocation is put to death, its thigh is taken, is rushed bone marrow cell with cold PBS with syringe
Go out.Obtained cell BM-Mac culture mediums【In 80% (v/v) DMEM complete mediums+20% (v/v) L929 cell culture
Clearly】, being incubated at culture dish, (density is 5 × 106/ ware, culture volume 10mL).Culture 3 days, BM- fresh addition 5mL
Mac culture mediums；Cultivate the 5th day, replace with the fresh BM-Mac culture mediums of 10mL.Cultivate the 7th day, collect cell, be incubated at and contain
10% (v/v) FBS and penicillin containing 100U/mL, 100 μ g/mL streptomysins DMEM complete mediums 6 orifice plates in, cell is close
Spend for 1.2 × 106/ hole (2mL culture mediums), cultivate 24 hours.Then liquid is changed, the DMEM for adding the LPS containing 500ng/mL is trained completely
Support base (2mL) pretreatment processing 4 hours, activate BMDM cells, then through containing various concentrations lamp-dish flower acetic (0.1,0.2,
Culture medium (1.2mL) 0.4mmol/L) is handled 1 hour, is finally added ATP containing 3mmol/L and is handled 30 minutes.
(2) in supernatant caspase-1 and IL-1 β detection method：
Cell is drawn culture supernatant 1.2mL, simply centrifuged through 300 × g 5 minutes, supernatant is transferred to separately after above-mentioned processing
In one centrifuge tube, 21 μ L 10% (w/v) sodium deoxycholate solution (final concentration of 0.15% (w/v)) is first added, after shaking up, then
Add 94 μ L 100% trichloroacetic acid (final concentration of 7.2% (w/v)) it is well mixed after, stayed overnight in 4 DEG C of protein precipitations.Then
Centrifuged 30 minutes through 14000 × g, abandon supernatant.Precipitation adds 700 μ L cold acetones, washes away the trichloroacetic acid in precipitation；14000×g
Centrifugation 5 minutes.Supernatant is abandoned, repeats to wash twice with above-mentioned cold acetone.After last time washing centrifugation, supernatant is abandoned；Make residual in precipitation
Remaining acetone volatilizees at normal temperatures, is finally dissolved with protein electrophoresis sample liquid.Boil 5 minutes, the albumen after above-mentioned processing, use
Western blotting method detects.
(3) IL-1 β contents in CBA kits detection supernatant：
BMDM cells are seeded in 24 orifice plates to (density is 1.5 × 105/ hole), after staying overnight, 500ng/mL LPS inductions 4
Hour, lamp-dish flower acetic (0.1,0.2,0.4mmol/L) effect 1 hour is added, then adding 3mmol/L ATP stimulates 1 hour,
Cell culture supernatant is finally collected, using CBA Mouse IL-1 β Flex Set kit measurement IL-1 β contents, it is operated
Step is carried out according to kit specification.Finally with machine testing on flow cytometer, data are carried out by CELLQuest softwares and obtained
Take and analyze.
(4) result：Inflammation corpusculum activates degree with caspase-1 (p10 fragments) activation levels and IL-1 β release reflections.
Caspase-1 (p10) and IL-1 β expression is activated in cells and supernatant to be increased and drops with the dosage of lamp-dish flower acetic
It is low.Illustrate that lamp-dish flower acetic (scutellarin) dose-dependently suppresses inflammation corpusculum in the BMDM cells that LPS+ATP is induced
Activation (Fig. 1).'beta '-tubulin is the internal reference albumen of electrophoresis loading in Fig. 1, illustrates that each swimming lane loading Tot Prot is equal.
Embodiment 2
(1) detection that ASC patches are formed：
BMDM cells are seeded in Glass bottom culture dish to (density is 5 × 105/ hole), after staying overnight, 500ng/mL LPS inductions
4 hours, add lamp-dish flower acetic 0.4mmol/L and act on 1 hour, then adding 3mmol/L ATP stimulates 1 hour, finally inhales and abandons
Its culture medium, add 1mL 4% (w/v) paraformaldehyde per hole, room temperature fixes 15 minutes, then adds 2mL cold methanols in -20 DEG C per hole
Under the conditions of penetrating 10 minutes, then closed 1 hour with confining liquid room temperature, add primary antibody (100 μ L/ holes), 4 DEG C of overnight incubations, after washing
Add CF568- goat rabbit-anti IgG and the CF488A- goat anti-mouse IgG (being purchased from Biotium companies of the U.S.) of no cross reaction,
Incubation at room temperature 1 hour, Hoechst 33342 contaminates core 10 minutes and lucifuge, Zeiss inverted fluorescence microscope are observed and taken pictures.
(2) ASC oligomer detects：
BMDM cells are seeded in 6 orifice plates to (density is 1 × 106/ hole), after staying overnight, 500ng/mL LPS inductions 4 are small
When, add lamp-dish flower acetic 0.4mmol/L and act on 1 hour, finally inhale and abandon supernatant, the PBS that 500 μ L precoolings are added per hole contains
0.5% (v/v) TritonX-100.At 4 DEG C, 6000 × g is centrifuged 5 minutes cell pyrolysis liquid, and supernatant is abandoned in suction, adds the PBS of precooling
Wash twice, be then resuspended in 200 μ L PBS, be directly added into suberic acid pair (N-hydroxy-succinamide ester), make its final concentration
For 2mmol/L, react 30 minutes at room temperature.In 4 DEG C through 6000 × g centrifuge 15 minutes Aspirate supernatants after, add 25 μ L 2 ×
Electrophoresis sample-loading buffer, boil 5 minutes, the albumen after above-mentioned processing, detected with western blotting method.
(3) result：Fig. 2 is that the mouse bone marrow cells source property macrophage that lamp-dish flower acetic (scutellarin) is induced LPS+ATP is thin
Born of the same parents (BMDM) immunofluorescence microscopy is analyzed ASC and NLRP3 distribution and the formation of ASC patches (ASC speck) and is immunized
The formation of engram analysis ASC oligomer.Thus scheme visible, lamp-dish flower acetic (scutellarin) can significantly suppress LPS+ATP
The formation of ASC patches and ASC oligomer in the BMDM cells of induction, further illustrate that lamp-dish flower acetic can suppress NLRP3 inflammation
The activation of disease corpusculum.
Embodiment 3
(1) cell culture and processing：
BMDM cells are inoculated in 24 orifice plates to (density is 1.5 × 105/ hole), after staying overnight, induced with 500ng/mL LPS
4 hours, add lamp-dish flower acetic (0.1,0.2,0.4mmol/L) effect 1 hour, then add 3mmol/L ATP stimulate it is 1 small
When, it is eventually adding PI (2 μ g/mL) and Hoechst 33342 (5 μ g/mL) and dyes 10 minutes at room temperature, is placed in Zeiss and is inverted fluorescence
Micro- Microscopic observation is simultaneously taken pictures.
(2) in supernatant HMGB1 detection method：
Cell is drawn culture supernatant 1.2mL, simply centrifuged through 300 × g 5 minutes, supernatant is transferred to separately after above-mentioned processing
In one centrifuge tube, 21 μ L 10% (w/v) sodium deoxycholate solution (final concentration of 0.15% (w/v)) is first added, after shaking up, then
Add 94 μ L 100% trichloroacetic acid (final concentration of 7.2% (w/v)) it is well mixed after, stayed overnight in 4 DEG C of protein precipitations.Then
Centrifuged 30 minutes through 14000 × g, abandon supernatant.Precipitation adds 700 μ L cold acetones, washes away the trichloroacetic acid in precipitation；14000×g
Centrifugation 5 minutes.Supernatant is abandoned, repeats to wash twice with above-mentioned cold acetone.After last time washing centrifugation, supernatant is abandoned；Make residual in precipitation
Remaining acetone volatilizees at normal temperatures, is finally dissolved with protein electrophoresis sample liquid.Boil 5 minutes, the albumen after above-mentioned processing, use
Western blotting method detects.
(3) result：, can be by PI stained positives after inducing cell Jiao dies, while can promote danger signal molecule HMGB1's
Release.Therefore, the emission levels of HMGB1 in supernatant are dyed and detected by PI, can analyze the situation that cell Jiao dies.Fig. 3 is
The suppression that mouse bone marrow cells source property macrophage (BMDM) cell Jiao that lamp-dish flower acetic (scutellarin) is induced LPS+ATP dies
Propidium iodide (PI) the fluorescent staining figure and immunoblotting assay figure of making.Red fluorescence (PI dyeing) represents that the cell is burnt
Die, cell Jiao that lamp-dish flower acetic (scutellarin) dose-dependently suppresses the BMDM cells of LPS+ATP inductions dies and extremely
Die releases of the mark HMGB1 in supernatant.Thus scheme it is visible, lamp-dish flower acetic pretreatment can dose-dependent inhibition ATP lure
Cell Jiao led dies, while HMGB1 emission levels increase with the dosage of lamp-dish flower acetic and reduce (Fig. 3 C) in supernatant.Fig. 3 C
Middle 'beta '-tubulin is the internal reference albumen of electrophoresis loading, illustrates that each swimming lane loading Tot Prot is equal.
Embodiment 4
(1) Bacteria Culture：
Bacillus coli DH 5 alpha shakes bacterium overnight (37 DEG C) in Luria Broth (LB) culture medium, and next day presses 10% (v/v) bacterium
Liquid continues to shake bacterium amplification 3 hours (37 DEG C) in fresh culture.Bacterium solution after amplification centrifuges 10 points under 1824 × g centrifugal force
Clock, after PBS washings, it is standby to be resuspended in PBS solution.Bacterial density micro ultraviolet specrophotometer (Nanodrop 2000；
Thermo Scientific) measure；Corresponding CFU (CFU) is coated with culture by LB agarose plates and determined.
(2) zoopery：
C57BL/6 mouse, 6-8 weeks, female, purchased from Nanfang Medical Univ's Experimental Animal Center, in this laboratory adaptability
After cultivating one week, by the dosage of 100mg/kg and 200mg/kg body weight by the pre- gavage of PBS solution containing lamp-dish flower acetic, 3 hours
Afterwards, intraperitoneal inoculation Escherichia coli (2.5 × 109CFU/ is only), it is above-mentioned containing lamp-dish flower acetic to mouse stomach again after 1 hour
PBS solution.In ensuing 120 hours, mouse survival situation is observed and recorded, and draws mouse Kaplan-Meier within every 6 hours
Survivorship curve analyzes survival rate.The variance analysis of mouse survival rate is examined using Long-rank, P<0.05, which is considered as difference, statistics
Meaning.
(3) result：
Coli-infection, mouse can be caused because of infection and dead；And lamp-dish flower acetic (scutellarin) can effectively drop
The death rate (Fig. 4) of low abdominal cavity bacterial infection mouse.*P<0.05；**P<0.01.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (9)
1. lamp-dish flower acetic is preparing the application in treating NLRP3 relevant disease medicines.
2. lamp-dish flower acetic according to claim 1 is preparing the application in treating NLRP3 relevant disease medicines, its feature
It is：Described NLRP3 relevant diseases are autoimmunity disease caused by acquired diseases associated with inflammation and heredity NLRP3 mutation,
Heredity Cryopyrin associated period heat pyrexia syndromes, infective inflammation disease, and/or nerve degenerative diseases.
3. lamp-dish flower acetic according to claim 2 is preparing the application in treating NLRP3 relevant disease medicines, its feature
It is：
Described acquired diseases associated with inflammation and heredity NLRP3 mutation caused by autoimmunity disease be obesity, diabetes B,
Atherosclerosis and/or gout；
Described infective inflammation disease is pyemia, endotoxemia and/or cholestasis；
Described nerve degenerative diseases are multiple sclerosis, Alzheimer disease and/or Parkinson's disease.
4. lamp-dish flower acetic is preparing the corpusculum activation of suppression NLRP3 inflammation, suppress cell Jiao and die, suppress caspase-1 activation, press down
IL-1 β secretions processed, suppress HMGB1 secretions, suppress ASC patches and are formed, and suppress ASC oligomer and are formed, anti-systemic bacterial infection,
And/or the application in bacteria resistance medication for treating pyemia.
5. application according to claim 4, it is characterised in that：Described systemic bacterial infection is abdominal cavity bacterial infection.
6. the application according to claim 4 or 5, it is characterised in that：Described bacterium is gram negative pathogenic bacteria.
7. the application according to claim 4 or 5, it is characterised in that：Described bacterium is bacillus coli DH 5 alpha.
8. according to the application described in any one of claim 1~7, it is characterised in that：Described medicine includes pharmaceutically acceptable
Auxiliary material.
9. according to the application described in any one of claim 1~7, it is characterised in that：The formulation of described medicine is tablet, particle
Agent, capsule, dripping pill, sustained release agent, oral liquid or injection.
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| CN112245427A (en) * | 2020-10-28 | 2021-01-22 | 上海市第一人民医院 | Application of CY-09 in preparation of medicine for resisting osteolysis induced by artificial joint wear particles |
| CN112603924A (en) * | 2020-12-31 | 2021-04-06 | 大连医科大学 | Application of scutellarin in preparation of medicine for preventing and treating atherosclerosis |
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| CN110575451A (en) * | 2018-06-07 | 2019-12-17 | 云南中医学院 | application of tetraacetylscutellarin aglycone in medicine for treating and preventing sepsis |
| CN110368495A (en) * | 2019-04-16 | 2019-10-25 | 中山大学附属第一医院 | Application of the NLRP3/ASC inflammation corpusculum as treatment recurrent ischemia headstroke target spot |
| CN109908166A (en) * | 2019-04-30 | 2019-06-21 | 青岛大学附属医院 | Application of the scutellarin in preparation prevention/protection ischemia-reperfusion injury of kidney drug/pharmaceutical composition |
| CN110638824A (en) * | 2019-11-11 | 2020-01-03 | 南京中医药大学 | Application of scutellarin and analogue thereof as AQP8 regulator in preparation of medicine for treating gallstone |
| CN111358950A (en) * | 2020-03-20 | 2020-07-03 | 厦门大学 | Applications of Caspase-1 and ASC/Caspase-8 as targets |
| CN112245427A (en) * | 2020-10-28 | 2021-01-22 | 上海市第一人民医院 | Application of CY-09 in preparation of medicine for resisting osteolysis induced by artificial joint wear particles |
| WO2022143513A1 (en) * | 2020-12-29 | 2022-07-07 | 云南生物谷药业股份有限公司 | Oral preparation comprising erigeron breviscapus extract and preparation method therefor |
| CN112603924A (en) * | 2020-12-31 | 2021-04-06 | 大连医科大学 | Application of scutellarin in preparation of medicine for preventing and treating atherosclerosis |
| CN112941074A (en) * | 2021-01-20 | 2021-06-11 | 青岛大学附属医院 | Application of MicroRNA-302c-3p as NLRP3 inhibitor |
| CN112941074B (en) * | 2021-01-20 | 2022-06-28 | 青岛大学附属医院 | Application of MicroRNA-302c-3p as NLRP3 inhibitor |
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