CN108420826B - Application of lycium barbarum polysaccharide in preparation of medicine for treating or relieving depression and pharmaceutical composition containing lycium barbarum polysaccharide - Google Patents
Application of lycium barbarum polysaccharide in preparation of medicine for treating or relieving depression and pharmaceutical composition containing lycium barbarum polysaccharide Download PDFInfo
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Abstract
The invention discloses application of lycium barbarum polysaccharide in preparing a medicine for treating or relieving depression. It can be used alone or in combination with other components to make composition or drug combination, and is safe and effective. The invention also discloses a pharmaceutical composition for treating or relieving depression, which comprises the following components in parts by mass: 10-30 parts of wolfberry polysaccharide, 2-8 parts of chlorogenic acid and 1-3 parts of soybean isoflavone.
Description
Technical Field
The invention relates to a new application of lycium barbarum polysaccharide, in particular to an application of lycium barbarum polysaccharide in preparing a medicine for treating or relieving depression and a pharmaceutical composition containing lycium barbarum polysaccharide.
Background
Depression, also known as depressive disorder, is characterized clinically by a marked and persistent depression in the mood, the main type of mood disorder. The low mood is not matched with the situation in clinic, the depression of the mood can be from sultriness to sadness, and the self-declining depression and even the pessimism are taken away, and suicide attempts or behaviors can be caused; even the occurrence of stupor; in some cases, there is significant anxiety and motor agitation; in severe cases, psychotic symptoms such as hallucinations and delusions may occur. Each episode lasts at least 2 weeks, more than long, or even years, and most cases have a tendency to have recurrent episodes, most of which can be alleviated, and some of which can have residual symptoms or become chronic.
Depression is the fourth largest disease in the world, and is predicted to be the second largest by 2020; however, the medical treatment and prevention of the depression in China are in the situation of low recognition rate, hospitals above grade city have recognition rate less than 20%, and only less than 10% of patients receive related drug treatment; moreover, at the same time, the onset of depression (and suicidal events) has begun to trend toward a lower age (university, or even a group of primary and secondary school students). In conclusion, great importance is needed to science popularization, prevention and treatment of depression, and the prevention and treatment of depression are listed in the national mental health work key point.
At present, the western medicines for treating various kinds of depression mainly comprise tricyclic antidepressants (TCA for short, and divided into azines, prolines, dibenzos and the like), monoamine oxidase inhibitors (MAOI for short, such as isopropylhydrazine and the like) and 5-HT reuptake inhibitors (SSRI for short, such as fluoxetine and the like), but the western medicines generally have the defects of drug resistance, large toxic and side effects and the like, so that the search for the antidepressant medicines with safety, high efficiency and small side effects is particularly important, and especially the natural antidepressant medicines becomes a research hotspot in the field.
The medlar is a mature fruit of deciduous shrubs in solanaceae, is a traditional Chinese medicinal material in China, has the functions of nourishing and strengthening, replenishing vital essence and qi, dispelling wind, tonifying yang, strengthening muscles and bones and the like, and has the functions of reducing blood sugar, reducing blood fat, improving immunity, resisting oxidation, resisting fatigue and the like as the effective component of the medlar proved by modern scientific research. The Chinese wolfberry recorded in the dietetic herbal has the functions of strengthening muscle and tendon, resisting fatigue, dispelling wind, tonifying bones and muscles, benefiting people and removing consumptive disease. Related documents report the anti-fatigue data of wolfberry fruit as follows:
the influence of lycium barbarum polysaccharide on the anti-fatigue effect of mice is reported by the Louqiong, Hades and the like, and the experiment shows that the lycium barbarum polysaccharide is an effective component of the lycium barbarum anti-fatigue effect (the influence of the lycium barbarum polysaccharide on the anti-fatigue effect of the mice, academic newspaper of Hubei medical science, Oct1999, (20): 4); the hydrowarburg, fuwei loyalty, tan zong et al report that lycium barbarum polysaccharide has anti-fatigue effect (experimental study of lycium barbarum polysaccharide anti-fatigue effect, China pharmaceutical, August 2006, (6): 8); panjing, popjuan, Panxi Hua and the like report that the medlar has the functions of resisting fatigue, improving exercise endurance and improving immunity on mice (experimental research on the functions of resisting fatigue and enhancing immunity of medlar, Shanghai preventive medicine journal 2003, (15): 8).
Disclosure of Invention
In response to the above-mentioned deficiencies in the prior art, the present invention is directed to the discovery of drugs that may treat or alleviate depression.
One of the technical problems to be solved by the invention is to provide the application of the lycium barbarum polysaccharide in preparing a medicine for treating or relieving depression.
The second technical problem to be solved by the invention is to provide a pharmaceutical composition for treating or relieving depression, which is characterized by comprising the following components in parts by mass: 10-30 parts of wolfberry polysaccharide, 2-8 parts of chlorogenic acid and 1-3 parts of soybean isoflavone.
Preferably, the pharmaceutical composition further comprises: and 1-5 parts by mass of lentinan.
More preferably, the pharmaceutical composition further comprises: 0.5-1 part of schisandrin B.
Further preferably, the pharmaceutical composition further comprises: 0.5-1 part by mass of stachyose.
The lycium barbarum polysaccharide can be directly purchased or prepared by using a conventional method, and is preferably prepared by a water extraction and alcohol precipitation method.
Preferably, the preparation method of the lycium barbarum polysaccharide comprises the following steps:
(1) water extraction: adding deionized water into the wolfberry fruits according to the material-liquid ratio of 1: 5-1: 20(g/mL), soaking for 1-4 h, extracting for 1-2 h at the temperature of 60-90 ℃, centrifuging, collecting supernate, and concentrating to 1/10-1/15 of the original volume at the temperature of 50-70 ℃;
(2) alcohol precipitation: adding 90% ethanol into the concentrated solution, wherein the volume ratio of the concentrated solution to the ethanol is 1: 2-1: 5, standing for 12-24 hours, and separating out a precipitate to obtain the lycium barbarum polysaccharide.
Further preferably, the preparation method of the lycium barbarum polysaccharide comprises the following steps:
(1) water extraction: adding deionized water into the wolfberry fruits according to the material-liquid ratio of 1: 5-1: 20(g/mL), soaking for 1-4 h, extracting for 1-2 h at the temperature of 60-90 ℃, centrifuging, collecting supernate, and concentrating to 1/10-1/15 of the original volume at the temperature of 50-70 ℃;
(2) alcohol precipitation: adding 90% ethanol into the concentrated solution, wherein the volume ratio of the concentrated solution to the ethanol is 1: 2-1: 5, standing for 12-24 hours, and separating out a precipitate to obtain crude lycium barbarum polysaccharide;
(3) enzymolysis: adding water with the mass being 30-50 times of that of the crude wolfberry polysaccharide into the crude wolfberry polysaccharide to obtain a crude wolfberry polysaccharide aqueous solution, and then adding an enzyme, wherein the mass ratio of the crude wolfberry polysaccharide to the enzyme is 100: 1-50: 1; carrying out enzymolysis for 2-4 hours at 30-65 ℃; then treating the mixture in a water bath at 90-100 ℃ for 5-10 minutes, and stopping the enzymatic reaction to obtain enzymatic hydrolysate;
(4) alcohol precipitation: adding 90% ethanol into the enzymolysis liquid, wherein the volume ratio of the enzymolysis liquid to the ethanol is 1: 2-1: 5, standing for 12-24 hours, separating out a precipitate, centrifuging, and drying to obtain the lycium barbarum polysaccharide.
Wherein the content of the first and second substances,
the enzyme in the step (3) is one or more of pectinase, glucoamylase and papain, and preferably is a mixed enzyme of pectinase, glucoamylase and papain in a mass ratio of (3-5): 1-3): 1.
The drying in the step (4) can be freeze drying, spray drying, drying in a drying oven and the like.
The centrifugation in each step is 3000-4000 rpm.
The pharmaceutical composition can be obtained by directly and uniformly mixing the components.
The invention also provides application of the pharmaceutical composition in preparing a medicament for treating or relieving depression.
The preparation formulation of the medicament for treating or relieving depression can be tablets, granules, capsules, powder, pills, medicinal granules, oral liquid preparations and the like. The dosage forms may be prepared according to conventional techniques in the art.
The invention finds the new application of the lycium barbarum polysaccharide, and can be used for preparing the medicament for treating the depression independently or together with other natural extracted components. The Chinese wolfberry polysaccharide has the advantages that the symptom relieving condition of depression is improved, plant extracts are used, the Chinese wolfberry polysaccharide is almost free of toxic and side effects and drug resistance, the Chinese wolfberry polysaccharide has a synergistic effect on depression treatment when multiple components are compounded, and the Chinese wolfberry polysaccharide obtained by a specific enzymolysis method has a better effect on depression treatment.
Detailed Description
The present invention will be further described with reference to the following examples, which are intended to be illustrative only and not to be limiting of the invention in any way, and any person skilled in the art can modify the present invention by applying the teachings disclosed above and applying them to equivalent embodiments with equivalent modifications. Any simple modification or equivalent changes made to the following embodiments according to the technical essence of the present invention, without departing from the technical spirit of the present invention, fall within the scope of the present invention.
In the examples, glucoamylase, food grade, available from Jianglai Biotech GmbH, Shanghai, with an enzyme activity of 6 ten thousand/g, was used.
In the examples, pectinase is food grade, is purchased from Shanghai Jianglai biological science and technology limited, and has the enzyme activity of 10 ten thousand/g.
In the examples, the papain is food grade, is purchased from Shanghai Jianglai Biotech limited and has the enzyme activity of 10 ten thousand per gram.
EXAMPLE 1 preparation of Lycium barbarum polysaccharides (Water purification precipitation method)
The preparation method of the lycium barbarum polysaccharide comprises the following steps:
(1) water extraction: adding deionized water into fructus Lycii according to the material-liquid ratio of 1:5g/mL, soaking for 2h, extracting at 90 deg.C for 2h, centrifuging at 3500rpm, collecting supernatant, and concentrating at 60 deg.C to 1/10 of original volume;
(2) alcohol precipitation: adding 90% ethanol into the concentrated solution at a volume ratio of 1:5, standing for 12 hr, and precipitating to obtain fructus Lycii polysaccharide.
EXAMPLE 2 preparation of Lycium barbarum polysaccharides (Water purification and precipitation enzymolysis method)
(1) Water extraction: adding deionized water into fructus Lycii according to the material-liquid ratio of 1:5g/mL, soaking for 2h, extracting at 90 deg.C for 2h, centrifuging at 3500rpm, collecting supernatant, and concentrating at 60 deg.C to 1/10 of original volume;
(2) alcohol precipitation: adding 90% ethanol into the concentrated solution at a volume ratio of 1:5, standing for 12 hr, and precipitating to obtain fructus Lycii crude polysaccharide;
(3) enzymolysis: adding water with the mass 30 times of that of the crude wolfberry polysaccharide into the crude wolfberry polysaccharide to obtain a crude wolfberry polysaccharide aqueous solution, and then adding an enzyme, wherein the mass ratio of the crude wolfberry polysaccharide to the enzyme is 50: 1; enzymolysis at 37 deg.C for 2 hr; then treating in a water bath at 95 ℃ for 10 minutes, and terminating the enzymatic reaction to obtain enzymatic hydrolysate;
(4) alcohol precipitation: adding 90% ethanol into the enzymolysis liquid, keeping the volume ratio of the enzymolysis liquid to the ethanol at 1:5, standing for 12-24 hours, separating out a precipitate, centrifuging at 3500rpm, and drying to obtain the lycium barbarum polysaccharide.
And (3) the enzyme is a mixed enzyme with the mass ratio of pectinase, glucoamylase and papain being 5:1: 1.
Example 3 preparation of Lycium barbarum polysaccharides
Lycium barbarum polysaccharides were prepared according to the method of example 2, with the exception that: the enzyme in the step (3) is pectinase.
Example 4 preparation of Lycium barbarum polysaccharides
Lycium barbarum polysaccharides were prepared according to the method of example 2, with the exception that: the enzyme in the step (3) is glucoamylase.
Example 5 preparation of Lycium barbarum polysaccharides
Lycium barbarum polysaccharides were prepared according to the method of example 2, with the exception that: the enzyme in the step (3) is papain.
Example 6 preparation of Lycium barbarum polysaccharides
Lycium barbarum polysaccharides were prepared according to the method of example 2, with the exception that: the enzyme in the step (3) is a mixed enzyme with the mass ratio of pectinase to glucoamylase of 5: 2.
Example 7 preparation of Lycium barbarum polysaccharides
Lycium barbarum polysaccharides were prepared according to the method of example 2, with the exception that: and (3) the enzyme is a mixed enzyme with the mass ratio of pectinase to papain being 5: 2. .
Test example 1 Effect of Lycium Bararum polysaccharides on Depression behavior of mouse as behavioral despair model
1 materials and methods
1.1 materials
1.1.1 major drugs and reagents
Lycium barbarum polysaccharides provided in examples 1-7;
fluoxetine hydrochloride capsules, shanghai, zhongxi pharmaceutical ltd.
1.1.2 Experimental animals
The cleaning agent Kunming mouse is half of male and female, the weight is 18-22g, and the cleaning agent is provided by the experimental animal center of Jilin university. Animals are raised in cages, the circadian rhythm is kept for 12h, the room temperature is 24 +/-1 ℃, and the animals are fed with free drinking water.
1.2 methods
1.2.1 grouping and administration
The mice were randomly divided into 9 groups of 10 mice each, each half of the males and females, and the divided and administered doses were: negative control group (physiological saline, 0.1ml/kg wt., intragastric administration), positive control group (fluoxetine, 10mg/kg wt., intragastric administration), wolfberry polysaccharide of example 1-7 (0.2g/kg wt., intragastric administration), according to the administration dose and 0.1ml/10g administration volume, each mouse administration range is between 0.2 ml-0.8 ml, dissolved with physiological saline, prepared into solution of appropriate concentration.
1.2.2 forced swimming test
After the last administration for 1h, the animals are respectively put into a bucket with the water temperature of 23-25 ℃ so that hind limbs of the animals can not support the body by the bottom of the bucket. The experiment consisted of two parts: pre-swimming for 15min, observing swimming behavior within 6min after 24h, and recording the immobility time within 5 min.
1.2.3 Tail suspension immobilization experiment
After the last administration for 1h, the part which is about 1cm away from the tail tip of the mouse is pasted on one end of the thin line by using a medical adhesive tape, the other end of the thin line is connected with a tension transducer which is connected with the thin line, the head of the mouse is about 5cm away from the table top, 2 mice are hung once, the middle of the mouse is separated by white paper, the hanging time is 6min, and the cumulative immobile time of the mouse within 5min after statistics.
2 results
2.1 mouse forced swimming test results
The results are shown in table 1, after 7 days of continuous administration, the lycium barbarum polysaccharide and fluoxetine positive control group can significantly reduce the immobility time of the mice in the forced swimming experiment compared with the negative control group. Example 2 compared with example 1, the use of lycium barbarum polysaccharide subjected to enzymatic hydrolysis and alcohol precipitation has a better effect, probably because the composition of lycium barbarum polysaccharide subjected to enzymatic hydrolysis is more favorable for treating depression or the molecular weight of lycium barbarum polysaccharide subjected to enzymatic hydrolysis is favorable for improving bioavailability. The enzymolysis is carried out by using three enzymes of pectinase, glucoamylase and papain, the effect is most obvious, and the effect is relatively poor when the glucoamylase or the papain is used alone.
Table 1 effect on immobility time in forced swimming of mice (n ═ 10, x ± SEM)
| Group of | Motionless time/s |
| Negative control group | 126.7±11.9 |
| Positive control group | 88.1±9.6 |
| Example 1 Lycium barbarum polysaccharides group | 118.9±11.0 |
| Example 2 Lycium barbarum polysaccharides group | 83.9±6.9 |
| Example 3 Lycium barbarum polysaccharides group | 95.5±8.7 |
| Example 4 Lycium barbarum polysaccharides group | 108.2±10.3 |
| Example 5 Lycium barbarum polysaccharides group | 112.4±7.2 |
| Example 6 Lycium barbarum polysaccharides group | 86.9±7.2 |
| Example 7 Lycium barbarum polysaccharides group | 90.5±8.1 |
2.2 mouse Tail suspension immobility test results
The results are shown in table 2, after 7 days of continuous administration, the lycium barbarum polysaccharide groups and fluoxetine positive control groups in examples 1-7 can obviously reduce the tail suspension immobility time of mice compared with the negative control group. Example 2 compared with example 1, the use of lycium barbarum polysaccharide subjected to enzymatic hydrolysis and alcohol precipitation has a better effect, probably because the composition of lycium barbarum polysaccharide subjected to enzymatic hydrolysis is more favorable for treating depression or the molecular weight of lycium barbarum polysaccharide subjected to enzymatic hydrolysis is favorable for improving bioavailability. The enzymolysis is carried out by using three enzymes of pectinase, glucoamylase and papain, the effect is most obvious, and the effect is relatively poor when the glucoamylase or the papain is used alone.
Table 2 effect on immobility time of mice tail suspension (n ═ 10, x ± SEM)
| Group of | Motionless time/s |
| Negative control group | 119.4±10.5 |
| Positive control group | 87.1±7.2 |
| Example 1 Lycium barbarum polysaccharides group | 106.1±10.5 |
| Example 2 Lycium barbarum polysaccharides group | 81.4±7.1 |
| Example 3 Lycium barbarum polysaccharides group | 88.5±6.2 |
| Example 4 Lycium barbarum polysaccharides group | 114.1±8.0 |
| Example 5 Lycium barbarum polysaccharides group | 106.4±7.9 |
| Example 6 Lycium barbarum polysaccharides group | 85.7±8.2 |
| Example 7 Lycium barbarum polysaccharides group | 84.6±7.4 |
EXAMPLES 8 to 13 preparation of pharmaceutical compositions
Weighing the components according to the mass parts in the following table, uniformly mixing, preparing the pharmaceutical composition of the embodiment 8-13, and meanwhile, setting 100% of lycium barbarum polysaccharide, 100% of chlorogenic acid and 100% of soybean isoflavone as comparative examples 1-3. The lycium barbarum polysaccharides in the table used the product prepared in example 2:
TABLE 3 composition of the pharmaceutical compositions
| Lycium barbarum polysaccharides | Chlorogenic acid | Soy isoflavone | Lentinan | Schisandra chinensis B element | Stachyose | |
| Example 8 | 10 parts by mass | 8 parts by mass | 1 part by mass | |||
| Example 9 | 30 parts by mass | 2 parts by mass | 3 parts by mass | |||
| Example 10 | 15 parts by mass | 5 parts by mass | 1 part by mass | |||
| Example 11 | 15 parts by mass | 5 parts by mass | 1 part by mass | 1 part by mass | ||
| Example 12 | 15 parts by mass | 5 parts by mass | 1 part by mass | 1 part by mass | 0.5 part by mass | |
| Example 13 | 15 parts by mass | 5 parts by mass | 1 part by mass | 1 part by mass | 0.5 part by mass | 0.5 part by mass |
| Comparative example 1 | 100％ | |||||
| Comparative example 2 | 100％ | |||||
| Comparative example 3 | 100％ |
Test example 2 Effect on drug-induced antidepressant model mouse behavior
1 materials and methods
1.1 materials
5-hydroxytryptophan (5-Hydroxy-DL-tryptophan, 5-HTP), Sigma;
reserpine, Sgima corporation;
the pharmaceutical composition of examples 8-13;
the medicament of comparative examples 1-3;
fluoxetine hydrochloride capsules, shanghai, zhongxi pharmaceutical ltd.
1.2 Experimental animals
The cleaning agent Kunming mouse, male and female, with a weight of 18-22g, is provided by the experimental animal center of Jilin university, and is raised in cages, and the daily rhythm is maintained for 12h, the room temperature is 24 +/-1 ℃, and the Kunming mouse can be freely drunk by drinking water.
1.1.3 Main Experimental instruments
Electronic balance (sidoris, germany BP 221S).
1.2 Experimental methods
1.2.15-HTP induced head-shaking experiment
Mice were randomly divided into 11 groups of 10 mice each, each half of males and females, and the divided and administered doses were: a negative control group (physiological saline, 0.1ml/kg wt., intragastric administration), a positive control group (fluoxetine, 10mg/kg wt., intragastric administration), a pharmaceutical composition group of examples 8-13 (0.2g/kg wt., intragastric administration), a pharmaceutical group of comparative examples 1-3 (0.2g/kg wt., intragastric administration), wherein the fluoxetine and the pharmaceutical composition or the drug are administered according to the administration dose and the administration volume of 0.1ml/10g, the administration range of each mouse is between 0.2ml and 0.8ml, and the solution is dissolved by physiological saline to be prepared into a solution with a proper concentration.
After the animals in each group are continuously dosed for 7 days, 100mg/kg wt.5-HTP is injected into the abdominal cavity after 1h of the last dose, continuous observation is started for 15min after 5min of dosing, and the times of head swinging of each group of mice are recorded.
1.2.2 reserpine Induction test
Animals were dosed and divided as described in 1.2.1 of test example 2, and a group of blank groups (10 mice) was added, and after 7 days of continuous dosing of each group of animals, 4mg/kg wt. reserpine was intraperitoneally injected into each group after 1 hour of the last dosing of each group except for the blank group of distilled water with the same volume as that of the stomach, and the observation of the drop in body temperature was performed.
2 results of the experiment
2.1 Effect on 5-HTP Induction experiments
The results are shown in table 4, and compared with the negative control group, the positive control group with fluoxetine dosage and the pharmaceutical composition of the present invention can significantly increase the number of times of head shaking of mice.
Table 4 effect on 5-HTP induction experiments (n ═ 10, x ± SEM)
2.2 Effect on reserpine Induction experiments
The results are shown in table 5, compared with the negative control group, the positive control group with fluoxetine dosage and the pharmaceutical composition of the invention can both significantly antagonize the effect of reserpine on inducing the body temperature drop of mice, which indicates that the pharmaceutical composition of the invention has an antidepressant effect, and the antidepressant effect is related to the influence on the central monoaminergic nervous system.
Table 5 effect on reserpine induction experiments (n ═ 10, x ± SEM)
Claims (3)
1. The pharmaceutical composition for treating or relieving depression is characterized by comprising the following components in parts by mass: 10-30 parts of lycium barbarum polysaccharide, 2-8 parts of chlorogenic acid, 1-3 parts of soybean isoflavone, 1-5 parts of lentinan, 0.5-1 part of schisandrin B and 0.5-1 part of stachyose.
2. The pharmaceutical composition of claim 1, wherein: the preparation method of the lycium barbarum polysaccharide comprises the following steps:
(1) water extraction: adding deionized water into the wolfberry fruits according to the material-liquid ratio of 1: 5-1: 20(g/mL), soaking for 1-4 h, extracting for 1-2 h at the temperature of 60-90 ℃, centrifuging, collecting supernate, and concentrating to 1/10-1/15 of the original volume at the temperature of 50-70 ℃;
(2) alcohol precipitation: adding 90% ethanol into the concentrated solution, wherein the volume ratio of the concentrated solution to the ethanol is 1: 2-1: 5, standing for 12-24 hours, and separating out a precipitate to obtain crude lycium barbarum polysaccharide;
(3) enzymolysis: adding water with the mass being 30-50 times of that of the crude wolfberry polysaccharide into the crude wolfberry polysaccharide to obtain a crude wolfberry polysaccharide aqueous solution, and then adding an enzyme, wherein the mass ratio of the crude wolfberry polysaccharide to the enzyme is 100: 1-50: 1; carrying out enzymolysis for 2-4 hours at 30-65 ℃; then treating the mixture in a water bath at 90-100 ℃ for 5-10 minutes, and stopping the enzymatic reaction to obtain enzymatic hydrolysate;
(4) alcohol precipitation: adding 90% ethanol into the enzymolysis liquid, wherein the volume ratio of the enzymolysis liquid to the ethanol is 1: 2-1: 5, standing for 12-24 hours, separating out a precipitate, centrifuging, and drying to obtain lycium barbarum polysaccharide; the enzyme in the step (3) is a mixed enzyme with the mass ratio of pectinase, glucoamylase and papain being (3-5): 1-3): 1.
3. Use of a pharmaceutical composition according to claim 1 for the manufacture of a medicament for the treatment or alleviation of depression.
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| CN112843221A (en) * | 2021-02-01 | 2021-05-28 | 宁夏杞肽科技有限公司 | Application of pharmaceutical composition in preparation of medicine for treating subliminal depression |
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