CN115089596A - New application of new agaro-oligosaccharide in preparing product for treating depression - Google Patents
New application of new agaro-oligosaccharide in preparing product for treating depression Download PDFInfo
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- CN115089596A CN115089596A CN202210698034.7A CN202210698034A CN115089596A CN 115089596 A CN115089596 A CN 115089596A CN 202210698034 A CN202210698034 A CN 202210698034A CN 115089596 A CN115089596 A CN 115089596A
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- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
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Abstract
The invention relates to a new application of new agaro-oligosaccharide in preparing a product for treating depression, the invention discovers that the new agaro-oligosaccharide has obvious effects on preventing and treating depression for the first time, and animal experiments verify that the new agaro-oligosaccharide has equivalent or even better curative effect than the drug approved in the market when being used for treating depression model mice. In addition, the invention discovers that the action mechanism of the new agaro-oligosaccharide for preventing and treating the depression at least comprises the following steps: improving the expression level of short-chain fatty acid in intestinal tract, improving the BDNF content in brain, and the like, which is consistent with the reported treatment mechanism of depression. The new agaro-oligosaccharide is used for preventing and treating depression, has the advantages of no side effect, high safety and the like, and has good application prospect.
Description
Technical Field
The invention relates to the technical field of biological medicines, in particular to a new application of new agaro-oligosaccharide in preparing a product for treating depression.
Background
Depression is a disease characterized primarily clinically by a marked and persistent depression in mood. According to the statistics of the world health organization, more than 3.2 million people worldwide suffer from depression. By 2030, depression is expected to be the leading factor in the global burden of disease. The core symptoms of depression include negative thought, anhedonia, anxiety and irritability, inattention, abnormal appetite and libido, abnormal sleep and tendency to self-destruction. Among them, anxiety is one of the most common co-morbid symptoms of depression. Depression has obvious biological factors, and the brain of a patient has important changes, such as imbalance of neurotransmitters in the brain, impaired nerve growth, reduced neural plasticity and abnormal neural circuits. Therefore, prevention, alleviation and treatment of depression have become an urgent global public health problem.
Various antidepressant drugs have been developed, including selective 5-hydroxytryptamine reuptake inhibitors, tricyclic drugs, monoamine oxidase inhibitors, etc., which can exert a certain therapeutic effect in the treatment of anxiety-complicated depression. Most of the chemically synthesized antidepressant drugs are neurotransmitter reuptake inhibitors, and due to complicated pharmacology and poor target specificity, drug dependent patients often show obvious toxic and side effects and serious adverse reactions such as liver and kidney function damage, hypertension, digestive tract abnormality, arrhythmia, dysmnesia, endocrine disorder and the like.
Prebiotics are substances that are not digested and absorbed by the host but selectively promote the metabolism and proliferation of beneficial bacteria in the body, including oligosaccharides, microalgae, protein hydrolysates, polysaccharides, natural plants. The prebiotics can be decomposed and absorbed by beneficial bacteria in intestinal tract, and promote growth and reproduction of the beneficial bacteria, thereby improving balance of intestinal flora. Research reports that prebiotics (oligosaccharides such as oligolactose, jerusalem artichoke oligosaccharide, morinda officinalis oligosaccharide and the like) have a certain treatment effect on depression. Therefore, the prebiotics has good application and development prospects in treatment of depression.
Disclosure of Invention
The invention aims to provide a new application of new agaro-oligosaccharide in preparing a product for treating depression, and the invention finds that the new agaro-oligosaccharide has the effects of preventing and treating depression in the deep research of the new agaro-oligosaccharide, and has the effect equivalent to or better than that of a positive control through the verification of animal experiments.
The new agaro-oligosaccharide is obtained by degrading agar (from asparagus, laver, agar and the like) by a biological method or a chemical method, consists of a unit of new agarobiose, and has the polymerization degree of 2-20. As a prebiotic, the new agaro-oligosaccharide has small molecular weight and good water solubility, and researches report that the new agaro-oligosaccharide has various physiological activities such as anti-tumor, anti-inflammatory, anti-oxidation, anti-obesity, anti-diabetes and the like, and has good application value in the fields of food, cosmetics, medicines and the like, but the application in the aspect of anti-depression is not reported. The research team of the invention establishes a large-scale preparation technology of the new agaro-oligosaccharide by the enzymolysis method, establishes a first new agaro-oligosaccharide enzymolysis production line in China, and realizes the mass production of the new agaro-oligosaccharide. The activity of the new agaro-oligosaccharide is deeply researched at the cellular and animal level, and the new agaro-oligosaccharide is found to have obvious effect on preventing and treating depression for the first time.
To this end, the present invention provides, in a first aspect, the use of a novel agaro-oligosaccharide for the preparation of a product for the prevention and/or treatment of depression, wherein said novel agaro-oligosaccharide is used as an active ingredient.
Further, the new agaro-oligosaccharide comprises one or the combination of more than two of new agarobiose, new agarotetraose, new agarohexaose, new agarooctaose, new agarodecose and new agarododecaose.
In some embodiments, the neoagaro-oligosaccharide comprises neoagarotetraose and neoagarohexaose in a mass ratio of 5 to 6:3 to 4, preferably 6: 4.
Furthermore, the product for preventing and/or treating depression contains an effective dose of neoagaro-oligosaccharide.
Furthermore, the product for preventing and/or treating depression also comprises pharmaceutically acceptable auxiliary materials.
Further, the product for preventing and/or treating depression is an oral product.
Further, the product for preventing and/or treating depression is a medicament, a health product or a food.
Furthermore, the product for preventing and/or treating depression is a medicament, and the dosage form of the medicament comprises granules, capsules, tablets, powder, oral liquid, suspension, emulsion and the like.
In some embodiments, the product is prepared by dissolving the novel agaro-oligosaccharides in physiological saline.
Further, the product for preventing and/or treating depression also comprises other components for preventing and/or treating depression besides the new agaro-oligosaccharide.
Further, the component for preventing and/or treating depression other than the novel agaro-oligosaccharide is selected from one or the combination of more than two of the following groups: oligolactose, jerusalem artichoke oligosaccharides, morinda officinalis oligosaccharides, monoamine oxidase inhibitor MAOI, tricyclic antidepressant TCA, selective serotonin reuptake inhibitor SSRI, serotonin and norepinephrine reuptake inhibitor SNRI, noradrenergic and specific 5HT receptor antagonist NASSA, 5HT antagonist, melatonin, 5HT2C receptor antagonist.
Further, in the product for preventing and/or treating depression, the new agaro-oligosaccharide plays an active role comprising: improving the expression level of short-chain fatty acid in intestinal tract and improving the BDNF content in brain.
Further, the short chain fatty acids include caproic acid, valeric acid, butyric acid, isovaleric acid, propionic acid, isobutyric acid, and acetic acid.
In a second aspect of the invention, a product for the prevention and/or treatment of depression is provided, said product comprising an effective amount of neoagaro-oligosaccharide.
Further, the new agaro-oligosaccharide comprises one or a combination of more than two of new agarobiose, new agarotetraose, new agarohexaose, new agarooctaose, new agarodecose and new agarododecaose.
In some embodiments, the neoagaro-oligosaccharide comprises neoagarotetraose and neoagarohexaose in a mass ratio of 5 to 6:3 to 4, preferably 6: 4.
Furthermore, the product for preventing and/or treating depression also comprises pharmaceutically acceptable auxiliary materials.
Further, the product for preventing and/or treating depression is an oral product.
Further, the product for preventing and/or treating depression is a medicine, a health-care product or a food.
Furthermore, the product for preventing and/or treating depression is a medicament, and the dosage form of the medicament comprises granules, capsules, tablets, powder, oral liquid, suspension, emulsion and the like.
In some embodiments, the product is prepared by dissolving neoagaro-oligosaccharide in physiological saline.
Further, the product for preventing and/or treating depression also comprises other components for preventing and/or treating depression besides the new agaro-oligosaccharide.
Further, the component for preventing and/or treating depression other than the novel agaro-oligosaccharide is selected from one or the combination of more than two of the following groups: oligolactose, jerusalem artichoke oligosaccharides, morinda officinalis oligosaccharides, monoamine oxidase inhibitor MAOI, tricyclic antidepressant TCA, selective serotonin reuptake inhibitor SSRI, serotonin and norepinephrine reuptake inhibitor SNRI, noradrenergic and specific 5HT receptor antagonist NASSA, 5HT antagonist, melatonin, 5HT2C receptor antagonist.
Compared with the prior art, the technical scheme of the invention has the following beneficial effects:
the invention discovers that the new agaro-oligosaccharide has obvious effect on preventing and treating depression for the first time, and animal experiments prove that the curative effect of the new agaro-oligosaccharide is equivalent to or even better than that of the drug approved in the market when the new agaro-oligosaccharide is used for treating depression model mice; in addition, the invention also researches the action mechanism of the new agaro-oligosaccharide for preventing and treating the depression, and the discovered action mechanism comprises the improvement of the expression quantity of short-chain fatty acid in intestinal tracts, the improvement of the BDNF content in brain and the like, which is consistent with the treatment mechanism of the depression reported at present.
Drawings
Various additional advantages and benefits will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. In the drawings:
FIG. 1: the rate of change in body weight of each group of mice during the administration period;
FIG. 2 is a schematic diagram: in open field experiments, the residence time of mice in different administration groups in the central area;
FIG. 3: in forced swimming experiments, mice of different administration groups have immobility time in water;
FIG. 4: analyzing the total amount of short fatty acids in intestinal tracts of mice of different administration groups;
FIG. 5: analyzing the content of caproic acid in intestinal tracts of mice of different administration groups;
FIG. 6: analyzing the content of valeric acid in intestinal tracts of mice of different administration groups;
FIG. 7: analyzing the content of butyric acid in intestinal tracts of mice of different administration groups;
FIG. 8: analyzing the content of isovaleric acid in intestinal tracts of mice of different administration groups;
FIG. 9: analyzing the content of propionic acid in intestinal tracts of mice of different administration groups;
FIG. 10: the content analysis result of the isobutyric acid in the intestinal tract of the mice of different administration groups;
FIG. 11: analyzing the content of acetic acid in intestinal tracts of mice of different administration groups;
FIG. 12: and (3) detecting the BDNF content in the brains of mice of different administration groups.
Detailed Description
Exemplary embodiments of the present disclosure will be described in more detail below with reference to the accompanying drawings. While exemplary embodiments of the present disclosure are shown in the drawings, it should be understood that the present disclosure may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
Example 1 weight gain animal experiments
Male C57BL/6 mice at 5 weeks of age were randomly assigned to the placebo group, depression building group. The placebo group was fed water normally without any stimulation. Carrying out adaptive breeding on the depression molding groups for 1 week, then carrying out cage breeding, and molding by adopting a chronic restraint stress method (CRS): mice were placed in 50mL modified centrifuge tubes (holes drilled in the tube wall to facilitate breathing) and restrained daily for 4h for 28 days with no food intake during restraint.
After the Model is successfully formed, the blank Control group is a normal group (Control), and mice in the depression forming Model group are randomly divided into a Model group (Model), a new agar oligosaccharide Low dose group (Low), a new agar oligosaccharide High dose group (High), a Paroxetine treatment group (Paraoxetine) and a new agar oligosaccharide Composition group (Composition), wherein 12 mice are formed in each group. Normal group was filled with stomach sterile water; the model group is filled with stomach sterile water; a new agaro-oligosaccharide low-dose group intragastric new agaro-oligosaccharide aqueous solution (the new agaro-oligosaccharide aqueous solution is prepared by dissolving neoagaro tetrasaccharide and neoagaro hexaose in a mass ratio of 6:4 in sterile water), wherein the intragastric dose is 100mg/d/kg of new agaro-oligosaccharide; intragastric neoagaro-oligosaccharide aqueous solution (prepared by dissolving neoagaro-tetrasaccharide and neoagaro hexasaccharide in a mass ratio of 6:4 in sterile water) is perfused into a neoagaro-oligosaccharide high-dose group, and the intragastric dose is 200 mg/d/kg; the new agaro-oligosaccharide composition group comprises a normal saline solution of new agaro-oligosaccharide through gastric perfusion (the normal saline solution of the new agaro-oligosaccharide is prepared by dissolving neoagaro-tetrasaccharide and neoagaro-hexasaccharide in a mass ratio of 6: 4), and the gastric perfusion dosage is 200mg/d/kg of the new agaro-oligosaccharide; the paroxetine treatment group comprises a gastric lavage paroxetine solution, wherein the gastric lavage dose is 10mg/d/kg of paroxetine; the above groups were administered by gavage for 28 days.
The experimental results are as follows: after 28d of molding, the weight change of the molding group is obvious compared with that of a blank control group, and CRS molding can obviously relieve the weight increase of the mice, which indicates that molding is successful. The body weight of the mice was weighed every 7d during the administration experiment, and the rate of change of body weight was recorded as shown in fig. 1. According to the results shown in fig. 1, the group administered with new agaro-oligosaccharide can significantly reverse the slow weight gain caused by CRS after 28d administration, and the concentration of new agaro-oligosaccharide is dependent in a certain range.
Example 2 open field experiment
Open field experiments are classical behavioral experiments for evaluating the degree of anxiety in animals. The principle is based on the fact that animals are afraid of the nature of open space, activities of the animals are aversive, and in the face of new things, the animals generate curiosity to explore the new space, so that contradictory psychology is caused, and anxiety is generated.
Performing open field experiments on each group of mice subjected to administration for 28 days in example 1, setting tracking conditions, placing the mice in the middle of a central area, timing and shooting; the observation time was 4 min.
The residence time of the mice in the central area was counted, and the results are shown in FIG. 2. According to the results of fig. 2, both the new agaro-oligosaccharide High dose group (High) and the new agaro-oligosaccharide Composition group (Composition) significantly increased the residence time of CRS mice in the central region, indicating that the mice were more motivated to explore themselves and reversed the depressed-like behavior of CRS-induced mice.
Example 3 forced swimming test
The forced swimming test is mainly used for the research of antidepressant class. The apparatus is suitable for rats, mice or other laboratory animals, and by placing the laboratory animals in a confined environment (such as water) in which the animals struggle to attempt to escape and cannot escape, a non-avoidable compression environment is provided, and after a period of experimentation, the animals exhibit a typical "immobility" reflecting a so-called "behavioral despair condition".
Forced swimming experiments were performed on each group of mice after 28 days of administration in example 1, respectively, with tracking conditions set, the mice placed in water, timed, and photographed; the observation time was 4 min.
The immobility time of the mice was counted, and the results are shown in FIG. 3. According to the results in fig. 3, the group administered with the new agaro-oligosaccharide can effectively reduce the immobility time of CRS mice, and the concentration of the new agaro-oligosaccharide is dependent, which indicates that the mice have the desire to struggle and reverse the depression-like behavior of the mice caused by CRS.
Example 4 short chain fatty acid analysis
For each group of mice after 28 days of administration in example 1, the cecum contents were separately dissected, 300. mu.L of 50% acetonitrile was added to a sample of the cecum contents, vortexed for 1 minute, sonicated at 4 ℃ for 30 minutes and then centrifuged at 12000rpm for 15 minutes, followed by addition of 3-nitrophenylhydrazine (3-NPH, 200mM) and a solution of 1-ethyl- (3-dimethylaminopropyl) carbonyldiimine (EDC, 120mM, containing 6% pyridine) (acetonitrile: 3-NPM: EDC: 2:1:1, v/v/v), vortexed for 1 minute, and reacted at 40 ℃ for 1 hour with shaking every 5 minutes. After completion of the reaction, the supernatant was collected by centrifugation at 12000rpm for 15min at 4 ℃ and analyzed by LC-MS/MS by diluting 100 times with 50% acetonitrile water.
The total amount of eight fatty acids, caproic acid, valeric acid, butyric acid, isovaleric acid, propionic acid, isobutyric acid and acetic acid, and short chain fatty acids, in the cecal contents of the mice were analyzed, and the results are shown in fig. 4-11, wherein the graphs correspond to: FIG. 4 shows total short fatty acid, FIG. 5 shows caproic acid, FIG. 6 shows valeric acid, FIG. 7 shows butyric acid, FIG. 8 shows isovaleric acid, FIG. 9 shows propionic acid, FIG. 10 shows isobutyric acid, and FIG. 11 shows acetic acid. Compared with a blank Control group (Control), the content of the total short-chain fatty acid and various fatty acids in the caecum content of the Model group (Model) mice are reduced to different degrees, wherein the content of the total short-chain fatty acid, the content of caproic acid, valeric acid, butyric acid, propionic acid and isobutyric acid is obviously reduced.
In contrast, the caecum contents of mice administered with neoagaro-oligosaccharide were increased in the amounts of various short-chain fatty acids as compared with the Model group (Model), wherein the contents of valeric acid, propionic acid, isovaleric acid, isobutyric acid and hexanoic acid were even restored to the control group level. In addition, the contents of various short chain fatty acids in the mouse cecal contents of the High-concentration new agaro-oligosaccharide group (High) and the new agaro-oligosaccharide Composition group (Composition) are higher than those of the Low-concentration new agaro-oligosaccharide group (Low). In general, oral administration of a certain concentration of neoagaro-oligosaccharide can effectively reverse the reduction of short-chain fatty acids in feces of mice with depression induced by a restraint stress method.
Although the detailed mechanism is largely unknown, short chain fatty acids have been shown to be key molecules linking the gut and brain, contributing to the anti-inflammatory response of the central nervous system. In addition, short chain fatty acids play a key role in maintaining intestinal barrier, and physiological concentrations have a significant impact on intestinal barrier function. In the research of the invention, the new agaro-oligosaccharide can obviously reverse the reduction of the short-chain fatty acid level in the excrement of depressed mice, which is beneficial to improving the intestinal barrier and the central nervous system inflammation.
Example 5BDNF content determination
For each group of mice administered for 28 days in example 1, eye-picking and blood-drawing were performed, specifically as follows: collecting blood with a 1.5mL centrifuge tube, placing in a refrigerator at 4 ℃, centrifuging at 4 ℃ and 3000rpm for 20min, taking the supernatant, repeatedly centrifuging once, taking the supernatant, separately packaging and storing, and detecting the BDNF content with an ELISA kit. The detection results are shown in fig. 12.
Existing evidence has shown that BDNF (brain-derived neurotrophic factor) is one of the key factors in the pathogenesis of depression, and an increase in BDNF is a significant marker of the efficacy of antidepressants. According to the detection result, the BDNF content in the brain of the mouse is obviously reduced after CRS modeling, and the High-concentration new agaro-oligosaccharide group (High) and the new agaro-oligosaccharide combination group (Composition) can effectively reverse the phenomenon that the BDNF of the brain of the mouse is reduced due to CRS modeling, so that the depression behavior of the mouse caused by the CRS is improved.
The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any changes or substitutions that can be easily conceived by those skilled in the art within the technical scope of the present invention are included in the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (10)
1. Application of new agaro-oligosaccharide in preparing depression preventing and/or treating product, wherein the new agaro-oligosaccharide is used as an active ingredient.
2. The use of claim 1, wherein said neoagaro-oligosaccharide comprises one or a combination of two or more of neoagarobiose, neoagarotetraose, neoagarohexaose, neoagarooctaose, neoagarodecose, and neoagarodecabiose.
3. The use according to claim 1, characterized in that said product for the prevention and/or treatment of depression contains an effective amount of neoagaro-oligosaccharides.
4. The use of claim 1, wherein the product for the prevention and/or treatment of depression further comprises a pharmaceutically acceptable excipient.
5. The use according to claim 1, wherein the product for the prevention and/or treatment of depression is an oral product.
6. The use according to claim 1, wherein the product for preventing and/or treating depression is a medicament, health product or food.
7. The use according to claim 6, wherein the product for preventing and/or treating depression is a medicament in the form of granules, capsules, tablets, powders, oral liquids, suspensions or emulsions.
8. The use according to claim 1, wherein the product for the prevention and/or treatment of depression further comprises a component for the prevention and/or treatment of depression other than neoagaro-oligosaccharide;
preferably, the component for preventing and/or treating depression other than the novel agaro-oligosaccharide is selected from one or a combination of two or more of the following groups: oligolactose, jerusalem artichoke oligosaccharides, morinda officinalis oligosaccharides, monoamine oxidase inhibitor MAOI, tricyclic antidepressant TCA, selective serotonin reuptake inhibitor SSRI, serotonin and norepinephrine reuptake inhibitor SNRI, noradrenergic and specific 5HT receptor antagonist NASSA, 5HT antagonist, melatonin, 5HT2C receptor antagonist.
9. The use according to claim 1, characterized in that in said product for the prevention and/or treatment of depression, said new agaro-oligosaccharides exert an active action comprising: the expression level of short-chain fatty acid in intestinal tract is increased, and the BDNF content in brain is increased;
preferably, the short chain fatty acids include hexanoic acid, pentanoic acid, butanoic acid, isovaleric acid, propanoic acid, isobutyric acid, and acetic acid.
10. A product for preventing and/or treating depression is characterized in that the product for preventing and/or treating depression contains an effective dose of new agaro-oligosaccharide.
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