CN106176741A - Berberine application in preparation strengthens inflammation corpusculum pharmacological activation - Google Patents
Berberine application in preparation strengthens inflammation corpusculum pharmacological activation Download PDFInfo
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- A61K31/4375—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with one nitrogen as the only ring hetero atom ortho- or peri-condensed with heterocyclic ring systems the heterocyclic ring system containing a six-membered ring having nitrogen as a ring heteroatom, e.g. quinolizines, naphthyridines, berberine, vincamine
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Abstract
The invention discloses berberine application in preparation strengthens inflammation corpusculum pharmacological activation.Present inventors discovered that berberine can remarkably promote the inflammation corpusculum activation of the signal inductions such as ATP, improve activity and the secretion of ripe IL 1 β of caspase 1；Berberine can also promote the macrophage lethal effect to antibacterial；In abdominal cavity bacterial infection model, berberine can dramatically increase the survival rate of mice；Thus, berberine can strengthen the function of the inherent immunity cells such as macrophage by promoting the secretion of IL 1 β, significantly improve the resistance that antibacterial is infected by host.Therefore, berberine can be used for preparing enhancing inflammation corpusculum pharmacological activation, and this medicine can strengthen inherent immunity cell function, can treat the relevant diseases such as systemic bacterial infection.
Description
Technical field
The invention belongs to biomedicine and drug world, be specifically related to berberine and strengthen inflammation corpusculum pharmacological activation in preparation
In application.
Background technology
Berberine (Berberine), also known as berberine, is a kind of common isoquinoline alkaloid, molecular formula C20H18NO4.It
It is present in many plants that four sections ten such as Berberidaceae belong to, can extract from the plants such as Rhizoma Coptidis, Cortex Phellodendri, Radix Berberidis.Radix Berberidis Amurensis
Alkali is yellow needle-like crystals, fusing point 145 DEG C, is insoluble in water, can be dissolved in benzene, ether and chloroform.Common drug form is that hydrochloric acid is little
Bark of a cork tree alkali (structural formula is as follows), the dissolubility in water is smaller.Berberine is to dysentery bacterium, tubercule bacillus, pneumonia ball
The various bacteria such as bacterium, Bacillus typhi has inhibitory action, wherein the strongest to dysentery bacterium effect.Clinic is mainly used in treating antibacterial
Property dysentery and gastroenteritis.Berberine purposes in terms for the treatment of gastroenteritis, it is seen that publication file (application number
CN201210474880.7)。
Berberine hydrochloride (molecular weight 371.81g/mol)
Activating inflammation corpusculum is one of body most important inherent immunity defense mechanism, including: activate caspase-1/
Caspase-11, promotes that the inflammatory mediators such as interleukin-1 ' beta ' (IL-1 β) are ripe, discharges, and inducing cell Jiao die.Normal condition
Under, above-mentioned mechanism is conducive to strengthening the local inflammation reaction of infection site, raises the phagocytosiss such as more neutrophilic granulocyte and kills thin
Born of the same parents, promote the removing of pathogenic microorganism, the final alleviation accelerating inflammation and the recovery from illness of disease.Specifically, cell Jiao dies at machine
Having following effect in body anti-infectious immunity: first, research shows the expression of caspase-1/caspase-11 and the thin of mediation thereof
Jiao born of the same parents dies, and is conducive to suppressing to infect (including the chmice acute enteritis that dextran sulfate sodium is induced), accelerates tissue repair, and Jiao dies
The mice of defect (caspase-1/caspase-11 double knocks out), susceptible and dead.Secondly, cell Jiao dies beneficially intracellular
The release of pathogenic bacterium, and swallowed by other phagocyte, kill and remove, prevent pathogenic bacterium from the breeding of intracellular and persistently depositing
?.Mycobacterium tuberculosis, HIV etc. cause the pathogenic microorganism of mankind's major disease, the immunologic escape of complete set of having evolved
Mechanism；Promote that cell Jiao dies and be probably the potential strategy removing this type of pathogenic microorganism.3rd, it is that activation is huge that cell Jiao dies
One of phagocytal important escape mechanism: if macrophage sustained activation and do not occur Jiao to die, its immunity metabolic pathway
Change, cause lipid deposition, become foam cell or fatty macrophages, these cells secrete continuously inflammation because of
Son (endogenous pyrogen) and chemotactic factor, raising other immunocytes increases atherosclerosis and obesity to lesion tissue, general
The risk of the chronic inflammation diseases such as disease.Therefore, strengthen the activation of inflammation corpusculum of infection site and infection cell (or to be activated
Cell) Jiao die, promote ripe IL-1 β secretion, it is advantageously possible for strengthen local inflammation reaction, accelerate to remove pathogenic microorganism,
Promote reparation and the recovery from illness of disease of damage tissue.
In view of inflammation corpusculum path important function in inherent immunity is defendd, by strengthening the activation of inflammation corpusculum, will
Contribute to strengthening the inherent immunity cell function of host, improve anti-infection ability, for the treatment of the relevant diseases such as bacterial infection
New approach is provided.
At present, the berberine report in the function strengthening the activation of inflammation corpusculum not yet it is related to.
Summary of the invention
It is an object of the invention to the shortcoming overcoming prior art with not enough, it is provided that berberine strengthens inflammation corpusculum in preparation
Application in pharmacological activation.
The purpose of the present invention is achieved through the following technical solutions:
Berberine application in preparation strengthens inflammation corpusculum pharmacological activation.
The performance of described enhancing inflammation corpusculum activation includes following aspect: activate caspase-1, promotes inflammatory cytokine
Secretion, ripe, release, inducing cell Jiao die.
Described inflammatory cytokine is interleukin-1 ' beta ' (IL-1 β).
Described enhancing inflammation corpusculum pharmacological activation has enhancing inherent immunity cell function.
The application in preparation treatment systemic bacterial infection medicine of the described enhancing inflammation corpusculum pharmacological activation.
The application in preparation treatment chronic inflammation disease medicine of the described enhancing inflammation corpusculum pharmacological activation.
Described enhancing inflammation corpusculum pharmacological activation comprises acceptable adjuvant.
Described adjuvant be preferably slow releasing agent, excipient, filler, binding agent, wetting agent, disintegrating agent, absorption enhancer,
Absorption carrier, surfactant or lubricant.
Described enhancing inflammation corpusculum pharmacological activation also comprises other and plays the synergistic effective ingredient of compatibility.
Described enhancing inflammation corpusculum pharmacological activation can be tablet, granule, capsule, drop pill, slow releasing agent, oral liquid, note
Penetrate agent.
The present invention has such advantages as relative to prior art and effect: the present invention is first based on the present inventor
First find that berberine has promote IL-1 β secretion and strengthen the effect proposed invention creation of inherent immunity cell function.This
The inventor of invention finds: berberine can remarkably promote the macrophage proinflammatory inflammation factor IL-1 β of release that ATP induction LPS activates
With danger signal molecule HMGB1, strengthen its ability killing intracellular bacteria, effectively reduce the death that abdominal cavity bacterial infection causes.
The above-mentioned new activity of berberine and purposes, there is not yet open source literature report.Berberine activates by strengthening inflammation corpusculum, improves
IL-1 β secretes, thus strengthens the function of inherent immunity cell, infects clinical treatment systemic bacterial and chronic inflammation disease
Etc. significant, can be used for related drugs exploitation.
Accompanying drawing explanation
Fig. 1 is that the mouse macrophage J774A.1 inflammation corpusculum that LPS+ATP is induced by berberine (BBR) activates situation
Immunoblotting assay figure.
Fig. 2 is that the peritoneal macrophage that the mice thioglycolate salt (TG) that LPS+ATP is induced by berberine (BBR) is induced is scorching
Disease corpusculum activates the immunoblotting assay figure of situation.
Fig. 3 is that bone marrow derived macrophage (BMDM) the inflammation corpusculum that LPS+ATP is induced by berberine (BBR) activates
The immunoblotting assay figure of situation.
Fig. 4 is that berberine (BBR) promotes the mouse macrophage J774A.1 cell statistics to the lethal effect of phagocytosis antibacterial
Result figure；* P < 0.05 is represented.
Fig. 5 is the statistical result figure that berberine (BBR) reduces the mortality rate of abdominal cavity bacterial infection mice；* * represent P <
0.001。
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention do not limit
In this.
Berberine hydrochloride (purity > 98%) (production code member B3251), lipopolysaccharide (LPS) (name of product
Lipopolysaccharides from Escherichia coli 0111:B4, catalog number (Cat.No.): L4391), ATP (catalog number (Cat.No.):
A6419), dimethyl sulfoxide (DMSO) (catalog number (Cat.No.): D8418), Tween-80 (catalog number (Cat.No.): P8074) are purchased from Sigma-
Aldrich；In vitro cell experiment berberine is dissolved in DMSO, and experiment in vivo berberine (5mg/mL) is dissolved in containing 2% (v/v)
The PBS solution (pH=7.4) of Tween-80.The cell cultivation materials such as cell culture medium DMEM, hyclone (FBS) are
Thermo/Gibco Products.IL-1 β antibody (catalog number (Cat.No.): #12242) used, HMGB1 antibody (catalog number (Cat.No.): #3935), β-
Tubulin (tubulin) antibody (catalog number (Cat.No.): #2128) is Cell Signaling Technologies Products.Anti-
Caspase-1p10 antibody (catalog number (Cat.No.): sc-514) is Santa Cruz Biotechnology Products
Embodiment 1
(1) cell is cultivated and is processed
Mouse macrophage J774A.1 is purchased from Chinese Academy of Sciences's Shanghai cell bank, is incubated at containing 10% (v/v) hyclone
With containing 100U/mL penicillin, 100 μ g/mL streptomycins DMEM (complete medium) in.Treat that cell length, to exponential phase, connects
Planting in 6 orifice plates, cell density is 2.5 × 105/ hole (2mL culture medium), cultivates 24 hours.Then change liquid, add containing 500ng/
The complete medium (2mL) of mL LPS, pretreatment 4 hours, activate J774A.1 cell, then through the berberine containing variable concentrations
The DMEM culture medium (1.2mL) of (1.5,3,6 μm ol/L) and 0.1% (v/v) FBS processes 1 hour, finally adds containing ATP's
DMEM culture medium (the final concentration of 3mmol/L of ATP) 0.8mL processes 1 hour.
(2) detection method of caspase-1 and HMGB1 in supernatant
Cell, after above-mentioned process, is drawn culture supernatant, is centrifuged 5 minutes through 200 × g, and supernatant is transferred to another centrifuge tube
In, successively it is separately added into NaTDC (making its final concentration of 0.15% (w/v)) and trichloroacetic acid (TCA) (makes its final concentration
It is 7.2% (v/v)) after mix homogeneously, at 4 DEG C of protein precipitations overnight.Then it is centrifuged 30 minutes through 14000 × g, abandons supernatant.Heavy
Form sediment and add 700 μ L cold acetone (-20 DEG C), the TCA fully suspending, cleaning in precipitation.After Li Xin, abandon supernatant；Make in precipitation remaining
Acetone volatilize at normal temperatures, finally with protein electrophoresis sample liquid dissolve.Boil 5 minutes, the albumen after above-mentioned process, with exempting from
Epidemic disease immunoblot method detects.
(3) experimental result
Inflammation corpusculum activates degree and discharges reflection with pro-caspase-1 (45kDa) and HMGB1.Result as it is shown in figure 1,
In cells and supernatant, the expression of pro-caspase-1 and HMGB1 increases with the dosage of BBR and raises, berberine (BBR)
Dose-dependently promote the mouse macrophage J774A.1 cellular inflammation corpusculum activation of LPS+ATP induction.
Embodiment 2
(1) cell is cultivated and is processed
C57BL/6 mice, female, 6-8 week old, purchased from Nanfang Medical Univ's Experimental Animal Center.Adaptability cultivates one week
After, after the 1mL PBS solution lumbar injection containing 3% (w/v) thioglycolate salt (Thioglycollate, TG) stimulates 4 days, neck
Vertebra dislocation is put to death, and washes out its abdominal cavity free cell by PBS solution.It is incubated at containing 10%FBS with containing 100U/mL penicillin, 100 μ
In 6 orifice plates of the DMEM complete medium of g/mL streptomycin, cell density is 2.5 × 106/ hole (2mL culture medium), cultivates 24 little
Time.Then change liquid, addition complete medium (2mL) pretreatment containing 500ng/mL LPS 4 hours, activate peritoneal macrophage,
Culture medium (1.2mL) through the berberine (0.75,1.5,3.0 μm ol/L) containing variable concentrations processes 1 hour again, finally adds
ATP (making its final concentration of 2mmol/L) processes 30 minutes.
(2) detection method of caspase-1 and IL-1 β in supernatant
Cell after above-mentioned process, draw culture supernatant 1.2mL, be centrifuged 5 minutes through 200 × g, supernatant be transferred to another from
In heart pipe, it is initially charged 10% (w/v) sodium deoxycholate solution (final concentration of 0.15% (w/v)) of 21 μ L, after shaking up, adds
After 100% trichloroacetic acid (final concentration of 7.2% (the v/v)) mix homogeneously of 94 μ L, at 4 DEG C of protein precipitations overnight.Then warp
14000 × g is centrifuged 30 minutes, abandons supernatant.Precipitation adds 700 μ L cold acetone (-20 DEG C), washes away the trichloroacetic acid in precipitation；
14000 × g is centrifuged 5 minutes.Abandon supernatant, repeat to wash twice with above-mentioned cold acetone (-20 DEG C).After last washing is centrifugal, abandon
Supernatant；Make acetone remaining in precipitation volatilize at normal temperatures, finally dissolve by protein electrophoresis sample liquid.Boil 5 minutes, through above-mentioned
Albumen after process, detects with western blotting method.
(3) experimental result
Inflammation corpusculum activates degree and discharges reflection with caspase-1 (p10 fragment) activation levels and IL-1 β.Result such as Fig. 2
Shown in, the expression activating caspase-1 (p10) and IL-1 β in cells and supernatant increases with the dosage of BBR and raises,
Berberine (BBR) dose-dependently promotes the Turnover of Mouse Peritoneal Macrophages inflammation corpusculum activation that LPS+ATP induces.
Embodiment 3
(1) cell is cultivated and is processed
L cell L929 cell is purchased from Kunming Institute of Zoology, Chinese Academy of Sciences's cell bank.Cell culture processes
With reference to the operation sequence (Stanford) of Monack laboratory, by 1 × 108It is 500cm that individual cell is incubated at floor space2Three layers
In culture bottle, culture medium is DMEM complete medium (200mL).
The cultural method of the macrophage (BMDM) of bone marrow derived: C57BL/6 mice, 6-8 week, female, purchased from south
Medical university of side Experimental Animal Center., take its Thigh bone at cervical dislocation after death, with syringe, medullary cell DMEM is trained
Foster base is gone out.The cell BM-Mac culture medium obtained (train by 80% (v/v) DMEM complete medium+20% (v/v) L929 cell
Support supernatant), (density is 5 × 10 to be incubated at Micro-Organism Culture Dish6Individual cell/ware, culture volume is 8-10mL).Cultivate 3 days, add
Add the new culture medium of 10mL；Cultivate the 5th day, replace with above-mentioned fresh culture.Cultivate the 7th day, collect cell, be incubated at and contain
10% (v/v) FBS and containing 100U/mL penicillin, 100 μ g/mL streptomycins DMEM (complete medium) 6 orifice plates in, cell
Density is 2.3 × 106Individual/hole (2mL culture medium), cultivates 24 hours.Then change liquid, add the training completely containing 500ng/mL LPS
Support base (2mL) pretreatment 4 hours, activate BMDM cell, then through the berberine (0.75,1.5,3.0 μm ol/L) containing variable concentrations
Culture medium (1.2mL) process 1 hour, finally add ATP, make ATP at the final concentration of 2mmol/L of culture medium, process 30 points
Clock.
(2) detection method of caspase-1 and IL-1 β in supernatant
Cell after above-mentioned process, draw culture supernatant 1.2mL, be centrifuged 5 minutes through 200 × g, supernatant be transferred to another from
In heart pipe, it is initially charged 10% sodium deoxycholate solution (final concentration of 0.15% (w/v)) of 21 μ L, after shaking up, adds 94 μ L
100% trichloroacetic acid (final concentration of 7.2% (v/v)) mix homogeneously after, at 4 DEG C of protein precipitations overnight.Then through 14000 ×
G is centrifuged 30 minutes, abandons supernatant.Precipitation adds 700 μ L cold acetones, washes away the trichloroacetic acid in precipitation；14000 × g is centrifuged 5min,
Abandon supernatant, repeat to wash twice with above-mentioned cold acetone.After last washing is centrifugal, abandon supernatant；Acetone remaining in precipitation is made to exist
Volatilize under room temperature, finally dissolve by protein electrophoresis sample liquid.Boil 5 minutes, the albumen after above-mentioned process, use immunoblotting side
Method detects.
(3) experimental result
Inflammation corpusculum activates degree and discharges reflection with caspase-1 (p10 fragment) activation levels and IL-1 β.Result such as Fig. 3
Shown in, the expression activating caspase-1 (p10) and IL-1 β in cells and supernatant increases with the dosage of BBR and raises,
Berberine (BBR) dose-dependently promotes the Turnover of Mouse Peritoneal Macrophages inflammation corpusculum activation that LPS+ATP induces.
Embodiment 4
(1) antibacterial culturing
Escherichia coli (DH5 α) shake bacterium overnight (37 DEG C) in Luria Broth (LB) culture medium, and next day presses 10% bacterium solution
Fresh culture continues shake bacterium 3 hours (37 DEG C) of amplification.Bacterium solution after amplification is centrifuged 10 points under 1824 × g centrifugal force
Clock, after PBS washing, is resuspended in PBS solution standby.Bacterial density trace ultraviolet spectrophotometer (Nanodrop 2000；
Thermo Scientific) measure；Corresponding colony-forming units (colony forming units, CFU) is by tradition LB fine jade
The coating cultivation of lipolysaccharide flat board determines.
(2) bacterial killer
Mouse monokaryon macrophage J774 A.1 cell is incubated in the DMEM complete medium containing 10%FBS.Treat that cell is long
To exponential phase, planting in 24 orifice plates, cell density is 3.5 × 104/ hole (500 μ L culture medium), cultivates 24 hours.Change liquid
And after washing twice with DMEM, the culture fluid of antibiotic-free adds 2.8 × 105The escherichia coli alive of CFU, make cell
Phagocytosis antibacterial 2 hours；By the berberine (BBR, 1.5,3,6,12 μm ol/L) of variable concentrations) it is formulated in the culture fluid containing antibiotic
After middle continuation is cultivated 6 hours, change liquid and wash twice with the aforementioned DMEM without antibiotic；Finally, with 200 μ L concentration it is
0.5% (v/v) Triton X-100, cell lysis 5 minutes under room temperature, thus obtained cell pyrolysis liquid dilutes 10 times with PBS
After, take 100 μ L and carry out LB agar plate coating, be placed in 37 DEG C of incubators, after cultivating 24 hours, the bacterium colony grown is counted.Respectively
Between group, the variation analysis of clump count uses variance analysis and Tukey inspection, and P < 0.05 is considered as difference statistical significance.
(3) result
As shown in Figure 4, berberine (BBR) can promote the J774A.1 cell lethal effect to phagocytosis antibacterial to result.* P is represented
<0.05。
Embodiment 5
(1) antibacterial culturing
Bacillus coli DH 5 alpha shakes bacterium overnight (37 DEG C) in Luria Broth (LB) culture medium, and next day exists by 10% bacterium solution
Fresh culture continues shake bacterium 3 hours (37 DEG C) of amplification.Bacterium solution after amplification is centrifuged 10 minutes under 1824 × g centrifugal force,
After PBS washing, it is resuspended in PBS solution standby.Bacterial density trace ultraviolet spectrophotometer (Nanodrop 2000；Thermo
Scientific) measure；Corresponding colony-forming units (CFU) is determined by tradition LB agarose plate coating cultivation.
(2) zoopery
C57BL/6 mice (purchased from Nanfang Medical Univ's Experimental Animal Center), after this laboratory adaptability cultivates one week,
By the dosage of 100mg/kg body weight by after pre-for the PBS solution containing berberine gavage 3 days (once a day), intraperitoneal inoculation escherichia coli
(2×109CFU/ is only), in ensuing 96 hours, every 6 hours observed and recorded mouse survival situations, and draw mice
Kaplan-Meier survival curve analyzes survival rate.The variation analysis of mouse survival rate uses Long-rank inspection, and P < 0.05 regards
Statistical significance is had for difference.
(3) experimental result
Result is as it is shown in figure 5, coli-infection may result in mice death because of infection；And berberine (BBR) can be effective
Reduce the mortality rate of abdominal cavity bacterial infection mice.* * represents P < 0.001.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-described embodiment
Limit, the change made under other any spirit without departing from the present invention and principle, modify, substitute, combine, simplify,
All should be the substitute mode of equivalence, within being included in protection scope of the present invention.
Claims (10)
1. berberine application in preparation strengthens inflammation corpusculum pharmacological activation.
The berberine the most according to claim 1 application in preparation strengthens inflammation corpusculum pharmacological activation, it is characterised in that:
The performance of described enhancing inflammation corpusculum activation includes following aspect: activate caspase-1, promotes inflammatory cytokine secretion, becomes
Ripe, release, inducing cell Jiao die.
The berberine the most according to claim 2 application in preparation strengthens inflammation corpusculum pharmacological activation, it is characterised in that:
Described inflammatory cytokine is interleukin-1 ' beta '.
The berberine the most according to claim 1 application in preparation strengthens inflammation corpusculum pharmacological activation, it is characterised in that:
Described enhancing inflammation corpusculum pharmacological activation has enhancing inherent immunity cell function.
The berberine the most according to claim 1 application in preparation strengthens inflammation corpusculum pharmacological activation, it is characterised in that:
The application in preparation treatment systemic bacterial infection medicine of the described enhancing inflammation corpusculum pharmacological activation.
The berberine the most according to claim 1 application in preparation strengthens inflammation corpusculum pharmacological activation, it is characterised in that:
The application in preparation treatment chronic inflammation disease medicine of the described enhancing inflammation corpusculum pharmacological activation.
7. preparing, according to the berberine described in any one of claim 1～6, the application strengthened in inflammation corpusculum pharmacological activation, its
It is characterised by: described enhancing inflammation corpusculum pharmacological activation comprises acceptable adjuvant.
The berberine the most according to claim 7 application in preparation strengthens inflammation corpusculum pharmacological activation, it is characterised in that:
Described adjuvant is slow releasing agent, excipient, filler, binding agent, wetting agent, disintegrating agent, absorption enhancer, absorption carrier, table
Face activating agent or lubricant.
The berberine the most according to claim 7 application in preparation strengthens inflammation corpusculum pharmacological activation, it is characterised in that:
Described enhancing inflammation corpusculum pharmacological activation also comprises other and plays the synergistic effective ingredient of compatibility.
The berberine the most according to claim 7 application in preparation strengthens inflammation corpusculum pharmacological activation, its feature exists
In: described enhancing inflammation corpusculum pharmacological activation can be tablet, granule, capsule, drop pill, slow releasing agent, oral liquid, injection.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108371661A (en) * | 2018-01-19 | 2018-08-07 | 中国人民解放军军事科学院军事医学研究院 | Application of the thiolutin in inhibiting the activation of NLRP3 inflammation corpusculums |
| CN111012780A (en) * | 2019-12-23 | 2020-04-17 | 青岛市口腔医院 | Application of berberine in preventing and treating pulpitis |
| CN111012786A (en) * | 2019-12-16 | 2020-04-17 | 中山大学 | Small molecular compound for activating inflammatory bodies and application thereof |
-
2016
- 2016-07-22 CN CN201610585003.5A patent/CN106176741A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| LEE YS等: "Berberine, a natural plant product, activates AMP-activated protein kinase with beneficial metabolic effects in diabetic and insulin-resistant states", 《DIABETES》 * |
| LIAO KC等: "Activation of the Nlrp1b Inflammasome by Reduction of Cytosolic ATP", 《INFECTION AND IMMUNITY》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108371661A (en) * | 2018-01-19 | 2018-08-07 | 中国人民解放军军事科学院军事医学研究院 | Application of the thiolutin in inhibiting the activation of NLRP3 inflammation corpusculums |
| CN111012786A (en) * | 2019-12-16 | 2020-04-17 | 中山大学 | Small molecular compound for activating inflammatory bodies and application thereof |
| CN111012780A (en) * | 2019-12-23 | 2020-04-17 | 青岛市口腔医院 | Application of berberine in preventing and treating pulpitis |
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Application publication date: 20161207 |