CN109200054A - A kind of new opplication of glycyrrhizic acid - Google Patents
A kind of new opplication of glycyrrhizic acid Download PDFInfo
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- CN109200054A CN109200054A CN201811249531.9A CN201811249531A CN109200054A CN 109200054 A CN109200054 A CN 109200054A CN 201811249531 A CN201811249531 A CN 201811249531A CN 109200054 A CN109200054 A CN 109200054A
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Abstract
The present invention relates to field of medicaments, the in particular to new opplication of glycyrrhizic acid, especially inhibit the application in the relevant a variety of neurodegenerative disease drugs of neuroinflamation in preparation.The present invention, which is experimentally confirmed glycyrrhizic acid, effectively to inhibit the release of inflammatory factor, to reduce it to degree of injury caused by neuronal cell, improve the cognitive function of brain tissue.The above result shows that glycyrrhizic acid can be used for preparing neuroinflamation inhibitor and treatment medicine for senile dementia.
Description
Technical field
The present invention relates to field of medicaments, in particular to a kind of application of glycyrrhizic acid.
Technical background
The neurodegenerative diseases such as Neuroinflammation and Alzheimer's disease (Alzheimer ' s disease, AD) have
Close connection.For AD, the poly physical efficiency that A β is formed is thin by the small colloid of immunocyte-of various receptor activation brains
Born of the same parents and astroglia, and then cause the chronic inflammatory reaction of part.The Deiter's cells being activated can largely discharge rush
Inflammatory cytokine, such as TNF-α, IL-1 β and IL-6 etc. or other inflammatory mediators, as nitric oxide (NO), activating oxide,
Prostaglandin, proteolytic enzyme and some complement molecules.These pro-inflammatory cytokines or other inflammatory mediators can be in various degree
Neuron is caused to damage, or directly Induction of neuronal apoptosis.Meanwhile the neuron of apoptosis can further activate it is unactivated
Deiter's cells, formed vicious circle constantly cause the damage of neuron and the exception of synaptic function.
More and more researches show that: using neuroinflamation as target spot, by the excessive activation and rush that inhibit microglia
The release of inflammatory cytokine can effectively prevent the deterioration of AD disease.More and more neuroinflamation inhibitor are seen at present
Report, neuroinflamation inhibitor can intervene Neuroinflammation by different mechanism, adjust the activation of microglia, reduce
The release of pro-inflammatory cytokine, and from play the role of in varying degrees protect neuron.Therefore, exploitation has anti-neuritis
Related drugs, functional food and the health care product of disease are of great significance for treating or preventing nerve retrograde affection.
Summary of the invention
In view of this, the present invention provides a kind of new opplications of glycyrrhizic acid, i.e., in preparation treatment neurodegenerative disease
Application in drug.
In order to achieve the above-mentioned object of the invention, the present invention the following technical schemes are provided:
The present invention provides application of the glycyrrhizic acid in the drug of preparation treatment neurodegenerative disease.
The present invention provides application of the glycyrrhizic acid in the drug that preparation restores Glucose Metabolism in Brain Tissues level.
The present invention images the metaboilic level of test measurement brain tissue glucose by microPET/CT, and image results are shown,
The brain tissue of the model mice of lipopolysaccharides (lipopolysaccharide, LPS) intraperitoneal injection reduces the intake of glucose,
And the glucose metabolism level that the LPS model mice brain tissue of glycyrrhizic acid is given in stomach-filling is restored.Show that glycyrrhizic acid can be extensive
Multiple Glucose Metabolism in Brain Tissues is horizontal.
The present invention proves that glycyrrhizic acid can obviously reduce inflammatory factor TNF- in hippocampal tissue by immunohistochemical test
α and IL-1 β protein level.Therefore, the present invention also provides glycyrrhizic acids reduces TNF-α or IL-1 β egg in hippocampal tissue in preparation
Application and glycyrrhizic acid in the drug of white level reduce the drug of TNF-α or IL-1 β protein level in cerebral cortex in preparation
In application.
The present invention passes through water maze test on the neuroinflamation animal model that LPS intraperitoneal injection repeatedly, repeatedly is established
And immunohistochemical experiment, discovery glycyrrhizic acid can be effectively reduced the secretion level of inflammatory factor TNF-α and IL-1 β in brain tissue,
Inhibit the activation degree of microglia, and then alleviate neuroinflamation and damaged caused by neuronal cell, hence it is evident that improves nerve
The behaviouristics variation and relevant Glucose Metabolism in Brain Tissues that inflammation is induced are horizontal, the above results show that, glycyrrhizic acid can be controlled
The relevant neurodegenerative disease of neuroinflamation is treated or alleviated, can be used for preparing the relevant a variety of neurologicals of inhibition neuroinflamation
The drug of property disease.Therefore, the application the present invention provides glycyrrhizic acid in the drug of preparation treatment neurodegenerative disease.
Preferably, the neurodegenerative disease is the relevant disease of neuroinflamation.
Preferably, the relevant disease of the neuroinflamation includes Alzheimer disease.
In order to obtain better therapeutic effect, preferably, dosage 50mg/kg~100mg/kg of glycyrrhizic acid, i.e.,
The relevant neurodegenerative disease of neuroinflamation can effectively be treated or be alleviated to the glycyrrhizic acid that 50~100mg is taken in 1kg human body,
Glucose Metabolism in Brain Tissues level, which can effectively be restored, can be effectively reduced TNF-α or IL-1 β egg in hippocampal tissue and cerebral cortex
White level effectively inhibits the activation of microglia.
In above-mentioned application provided by the invention, preferably, the drug includes glycyrrhizic acid and pharmaceutically acceptable
Auxiliary material.
Preferably, the dosage form of drug is oral preparation or injection, wherein the dosage form of the drug is oral preparation,
The oral agents are dispersion powders, granule, pill, tablet, capsule, microcapsules, oral solution, pill or liposome.
The present invention has found that glycyrrhizic acid can effectively inhibit the activation and inflammatory factor of microglia by zoopery
Overexpression, reduces its damage to neuronal cell, improves the cognitive function of brain tissue, can be used for preparing inhibition neuroinflamation
The drug of relevant a variety of neurodegenerative diseases.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described.
Fig. 1 shows mouse microPET/CT imaging figure, wherein first row is Mice brain tissues cross section, and second row is mouse
Top view, third row are mouse side view；
Fig. 2 shows the Nissl's staining figure of Mice brain tissues paraffin section, wherein first row is Mice brain tissues coronal section,
40 times of amplification；Second row is hippocampus, and third row is cerebral cortex, second and third discharge is 400 times big；
Fig. 3 shows the immunohistochemistry figure of Mice brain tissues TNF-α and IL-1 β albumen, wherein first row is hippocampus, second
Row is cerebral cortex area；
Specific embodiment
The present invention provides a kind of new opplication of glycyrrhizic acid, those skilled in the art can use for reference present disclosure, suitably change
Into realization of process parameters.In particular, it should be pointed out that all similar substitutions and modifications are aobvious for a person skilled in the art
And be clear to, they are considered as being included in the present invention.Method and application of the invention is carried out by preferred embodiment
Description, related personnel can obviously not depart from the content of present invention, carried out in spirit and scope to method described herein and application
Change or appropriate changes and combinations, carry out implementation and application the technology of the present invention.
In the present invention, raw materials used and reagent is available on the market.
Wherein, experimental animal is purchased from Nanfang Medical Univ using male, the 6 week old C57 mouse of SPF grade of weight 20-22g
Experimental Animal Center, credit number SCXK (Guangdong) 2016-0041.Mouse feed is purchased from Nanfang Medical Univ's Experimental Animal Center.
Experimental animal is raised in cleaning grade laminar-flow rack, and environment temperature airconditioning control is 25 ± 1 DEG C, and relative humidity is 75 ± 10%, shines
Bright time 12h/d (7:00-19:00).
TNF-α and IL-1 β antibody are purchased from Beijing Bo Aosen Biotechnology Co., Ltd, and DAB color developing agent is purchased from DAKO company.
Below with reference to embodiment, the present invention is further explained:
1 glycyrrhizic acid of embodiment is to the LPS mouse AD symptom memory disorders induced and relevant Glucose Metabolism in Brain Tissues water
Flat influence.
Experimental animal is randomly divided into normal group, LPS group, positive drug group (LPS+TTP488,5mg/kg/day), glycyrrhizic acid
Low dose group (LPS+ glycyrrhizic acid, GA50,50mg/kg/day), glycyrrhizic acid high dose group (LPS+ glycyrrhizic acid, GA100,100mg/
Kg/day), every group 6.Each experimental group starts to be administered for 1 week before lps injection, and successive administration 2 weeks.Except for the normal group, remaining is each
Group investigates the memory function of mouse using water maze laboratory after continuous injection 1 week in the LPS that starts for the 7th day to be injected intraperitoneally of administration
Can, it is horizontal using micro-PET/CT visualization method measurement Glucose Metabolism in Brain Tissues.Experimental data indicates own with Mean ± SD
Experimental data is all made of that 19.0 software of SPSS is for statistical analysis, and parameter is tested with ANOVA between the group of each group mouse, and P <
0.05 is considered statistically significant.The results are shown in Table 1 for water maze laboratory, and micro-PET/CT image results are as shown in Figure 1.
Influence of 1 glycyrrhizic acid of table to the LPS mouse AD symptom memory disorders induced
By table 1 and Fig. 1 result it is found that LPS intraperitoneal injection has been obviously prolonged incubation period of the mouse in orientation navigation experiment,
Reduce mouse simultaneously and enter the number and time scale of target quadrant in space exploration experiment, and reduces Mice brain tissues
To the intake of glucose, lead to the impaired memory function of mouse.Continuous gavage gives the reality of high dose and low dosage glycyrrhizic acid
It tests a group Memory Function to be significantly improved, shorter latencies, the number and time scale into target quadrant increase.Even
Continuous stomach-filling, which gives high dose and low dosage glycyrrhizic acid, can significantly improve Mice brain tissues to the intake of glucose, wherein sweet
Oxalic acid high dose effect is suitable with positive drug group effect.Show that glycyrrhizic acid can significantly improve the Mice brain tissues of LPS induction
To the intake of glucose, the glucose metabolism for restoring brain tissue is horizontal.
Influence of 2 glycyrrhizic acid of embodiment to the LPS Mice brain tissues neure damage induced
Experimental group, dosage regimen are the same as embodiment 1.After mouse is administered, 0.7% chloral hydrate anesthesia is put to death, will be complete
Whole brain tissue takes out.48h is fixed in 4% paraformaldehyde, tap water is rinsed well, with the alcohol serial dehydration of various concentration.
Being dehydrated after this to be soaked in dimethylbenzene carries out transparent, and transparent sample is soaked 4h in the paraffin of molten state, embeds, cuts
Piece, dewaxing rehydration carry out Nissl's staining later, and optical microphotograph is under the microscope.As a result as shown in Figure 2.
By the result of Fig. 2 it is found that normally containing a large amount of Nissl body in the neuronal cell of group mouse, after injecting LPS,
The number of Nissl body substantially reduces；After giving glycyrrhizic acid processing, the number of Nissl body is dramatically increased, and shows that glycyrrhizic acid can
Neure damage caused by LPS is effectively relieved.
The influence of inflammatory reaction in the Mice brain tissues that 3 glycyrrhizic acid of embodiment induces LPS
Experimental group, dosage regimen are the same as embodiment 1.After mouse is administered, by brain after the execution of 0.7% chloral hydrate anesthesia
Tissue takes out, and isolates hippocampus and skin portion on ice.Using immunohistochemical experiment measurement mouse brain hippocampus in TNF-α and
The content of IL-1 β, experimental data are indicated with Mean ± SD, are carried out data processing using SPSS19.0 software, are examined using ANOVA
Statistical analysis is carried out, P < 0.05 shows that, with statistical difference, the results are shown in Table 2.
Immunohistochemical assay: taking out complete fresh brain tissue, after being placed in the fixation of 4% paraformaldehyde, starts after PBS washing
Paraffin section production: key step includes graded ethanol dehydration (70%, 95% and 100%) → gradient dimethylbenzene transparent (50%
With 100%) → waxdip (brain tissue is soaked in 40-50min in the paraffin of thawing) → embedding (paraffin of embedding is poured into solidifying
Gu (paraffin section cut is put in 62-65 DEG C of pre-temperature of baking oven and bakes piece 1h) → slice (being sliced with 5 μm of thickness) → roasting piece
Afterwards, dewax aquation in dimethylbenzene ethanol solution) (histotomy is placed in fills with citric acid pH6.0 antigen retrieval buffers to → antigen retrieval
Reparation box in, in carrying out antigen retrieval in micro-wave oven) → block endogenous peroxydase: 3% hydrogenperoxide steam generator, room temperature
It is protected from light and is incubated for 25min, then PBS washing → serum closing (is added dropwise BSA and is incubated for 30min) incubation of → primary antibody and (is added dropwise on slice
The TNF-α primary antibody that PBS is configured by a certain percentage, slice lie against 4 DEG C of overnight incubations in wet box) → secondary antibody incubation (shaking table washing
After primary antibody, the secondary antibody that kind corresponding to primary antibody is added dropwise covers tissue, is incubated at room temperature 50min) → DAB colour developing: shaking table washs secondary antibody
Afterwards, DAB developing solution is added dropwise in slice slightly drying, and developing time is controlled under microscope, and the positive is brown color, and tap water rinses slice eventually
Only colour developing → haematoxylin redye nucleus (shaking table wash secondary antibody after, be added dropwise haematoxylin dye liquor, redye 2min) → mounting (wash by shaking table
After washing 3 times, drying, neutral gum mounting) → microscopy takes pictures (slice in microscopically observation and acquire image), as a result such as Fig. 3
It is shown.
Table 2: inflammatory factor generates horizontal influence in the Mice brain tissues that glycyrrhizic acid induces LPS
Note: * indicates that there are statistical difference in P < 0.05 and normal group ratio；
# indicates that there are statistical difference in P < 0.05 and LPS group ratio.
By the result of table 2 and Fig. 3 it is found that after lps injection, cause inflammatory factor TNF-α, the overexpression of IL-1 β and small
The extensive activation of spongiocyte.Compared with LPS group, the TNF-α in hippocampus and cortex is can be significantly reduced in glycyrrhizic acid high dose group
With IL-1 β protein expression level (P < 0.05)；Glycyrrhizic acid low dose group can significantly reduce in IL-1 β and cortex in hippocampus
The expression (P < 0.05) of TNF-α, IL-1 β albumen, is effectively reduced the expression of TNF-α albumen in hippocampus.
The above results show that glycyrrhizic acid can effectively inhibit Activated Microglia and inflammatory factor TNF-α, IL-1 β
Secretion controls Neuroinflammation, to alleviate subsequent caused neure damage, improves memory disorders and motor coordination energy
Power can be used for the relevant a variety of neurodegenerative diseases of neuroinflamation.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, various improvements and modifications may be made without departing from the principle of the present invention, these improvements and modifications are also answered
It is considered as protection scope of the present invention.
Claims (9)
1. application of the glycyrrhizic acid in the drug that preparation restores Glucose Metabolism in Brain Tissues level.
2. glycyrrhizic acid reduces the application in hippocampal tissue in TNF-α or IL-1 β protein level drug in preparation.
3. glycyrrhizic acid reduces the application in cerebral cortex in TNF-α or IL-1 β protein level drug in preparation.
4. glycyrrhizic acid is in preparation prevention or the application of the drug for the treatment of neurodegenerative disease.
5. application according to claim 4, which is characterized in that the neurodegenerative disease is the relevant disease of neuroinflamation
Disease.
6. application according to claim 5, which is characterized in that the relevant disease of the neuroinflamation includes Alzheimer
Disease, Parkinson's disease, amyotrophic lateral sclerosis and multiple sclerosis.
7. according to the described in any item applications of claim 4 to 6, which is characterized in that the dosage of the glycyrrhizic acid is 50mg/
Kg~100mg/kg.
8. application according to claim 4, which is characterized in that the drug includes glycyrrhizic acid and pharmaceutically acceptable auxiliary
Material.
9. application according to claim 9, which is characterized in that the dosage form of the drug is oral preparation, the oral agents
For dispersion powders, granule, pill, tablet, capsule, microcapsules, oral solution, pill or liposome.
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| CN201811249531.9A CN109200054A (en) | 2018-10-25 | 2018-10-25 | A kind of new opplication of glycyrrhizic acid |
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| CN201811249531.9A CN109200054A (en) | 2018-10-25 | 2018-10-25 | A kind of new opplication of glycyrrhizic acid |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111195245A (en) * | 2020-01-21 | 2020-05-26 | 广东工业大学 | Application of elemene |
| CN112843113A (en) * | 2021-03-23 | 2021-05-28 | 南京医科大学 | A preparation for treating contact dermatitis |
| CN114159447A (en) * | 2021-11-05 | 2022-03-11 | 暨南大学 | Application of 18 beta-glycyrrhetinic acid in preparation of medicine for treating depression-related neuron protection |
| WO2022052016A1 (en) * | 2020-09-11 | 2022-03-17 | Liu Hsuan Miao | Pharmaceutical compositions and uses thereof in treating parkinson's disease |
| CN114712374A (en) * | 2022-02-11 | 2022-07-08 | 河北医科大学 | Use of glycyrrhizic acid and glycyrrhetinic acid as phosphodiesterase 4 inhibitor |
| CN115624543A (en) * | 2022-11-18 | 2023-01-20 | 北京中医药大学 | Medicine for treating migraine, pharmaceutical composition, preparation method and pharmaceutical application thereof |
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2018
- 2018-10-25 CN CN201811249531.9A patent/CN109200054A/en not_active Withdrawn
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111195245A (en) * | 2020-01-21 | 2020-05-26 | 广东工业大学 | Application of elemene |
| CN111195245B (en) * | 2020-01-21 | 2023-03-17 | 广东工业大学 | Application of elemene |
| WO2022052016A1 (en) * | 2020-09-11 | 2022-03-17 | Liu Hsuan Miao | Pharmaceutical compositions and uses thereof in treating parkinson's disease |
| CN112843113A (en) * | 2021-03-23 | 2021-05-28 | 南京医科大学 | A preparation for treating contact dermatitis |
| CN112843113B (en) * | 2021-03-23 | 2022-02-01 | 南京医科大学 | A preparation for treating contact dermatitis |
| CN114159447A (en) * | 2021-11-05 | 2022-03-11 | 暨南大学 | Application of 18 beta-glycyrrhetinic acid in preparation of medicine for treating depression-related neuron protection |
| CN114712374A (en) * | 2022-02-11 | 2022-07-08 | 河北医科大学 | Use of glycyrrhizic acid and glycyrrhetinic acid as phosphodiesterase 4 inhibitor |
| CN115624543A (en) * | 2022-11-18 | 2023-01-20 | 北京中医药大学 | Medicine for treating migraine, pharmaceutical composition, preparation method and pharmaceutical application thereof |
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