A small intestinal mucosa extract with preventive and therapeutic effects on depression

Abstract

The present invention belongs to the field of food and medicine, and specifically relates to a small intestinal mucosa extract having a preventive and therapeutic effect on depression. The present invention also provides a preparation method of the small intestinal mucosa extract and a health product, food or medicine containing the small intestinal mucosa extract. The small intestinal mucosa extract provided by the present invention has been found through pharmacological experiments on animal models to significantly improve the depression and anxiety state of depression model rats and increase the body weight of rats, and the therapeutic effect on depression is significantly better than the prior art.

Description

Technical Field

[0001] The invention belongs to the field of medicine, and in particular relates to a small intestinal mucosa extract having preventive and therapeutic effects on depression. Background Art

[0002] The main manifestations of depression are persistent low mood, slow thinking, lack of appetite and weight loss, decreased will and behavior, and in severe cases, suicidal attempts or behaviors. It is a widely prevalent mental illness in the world, with about 300 million people of all ages suffering from depression worldwide.

[0003] According to the World Health Organization, the incidence of depression increases by 113% each year and is expected to increase further, becoming the world's second largest disease by 2020. However, the widely used antidepressants currently have great limitations, and long-term use will have certain toxic side effects on the human body. The development of safe and effective drugs to prevent depression or to relieve depression through nutritional intervention is an urgent problem to be solved in today's society. Studies have shown that nutrients extracted from food, such as milk-derived active peptides, platycodon, tuckahoe polysaccharides, and small molecule compounds (CXZ-123), have a relieving effect on depression.

[0004] The small intestine is the most important part of the food digestion and absorption process. Jame (1997) pointed out that mucosal digestion is the final digestion stage of various nutrients, and the intestinal mucosa is the final digestion site of all nutrients, which plays a key role. In addition to its digestion and absorption functions, the small intestine is also an important endocrine organ. A variety of active substances have been isolated from the small intestine and its mucosa, including heparin sodium, various enzymes and antibacterial substances. The various biological activities of small intestinal mucosal extracts will surely become a hot topic for pharmacists in the future. Summary of the invention

[0005] The main purpose of the present invention is to provide a new use of small intestinal mucosa extract, that is, the small intestinal mucosa extract has a preventive and / or therapeutic effect on depression; in addition, the small intestinal mucosa extract has a significant improvement effect on the depression, anxiety state, learning and memory ability, and body weight of patients with depression.

[0006] The small intestinal mucosa extract of the invention is prepared from pig small intestinal mucosa through extraction and precipitation processes.

[0007] The extraction method may be one or more of water extraction and ultrasonic extraction; preferably, the extraction method is ultrasonic extraction.

[0008] The above-mentioned precipitation method can be one or more of ethanol precipitation method, graded precipitation method, and quaternary ammonium salt complexation method; preferably, the precipitation method is ethanol precipitation method.

[0009] Preferably, the above-mentioned small intestinal mucosa extract is prepared from porcine small intestinal mucosa by water extraction and alcohol precipitation process.

[0010] Specifically, the preparation method of the small intestinal mucosa extract comprises the following steps:

[0011] (1) Extraction: Take the small intestinal mucosa, crush it, add water and perform ultrasonic extraction, filter and obtain the water extract.

[0012] (2) Alcohol precipitation: add ethanol to the aqueous extract obtained in step (1), and filter to obtain a precipitate.

[0013] (3) Freeze-drying: The precipitate obtained in step (2) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0014] Wherein, the small intestinal mucosa in step (1) is selected from the small intestinal mucosa of mammals; preferably, the small intestinal mucosa is porcine small intestinal mucosa.

[0015] Preferably, the volume mass ratio of water used in ultrasonic water extraction to small intestinal mucosa in step (1) is 1:1-100:1 (ml / g); further preferably, the volume mass ratio of water used to small intestinal mucosa is 80:1 (ml / g).

[0016] Preferably, ethanol is added in step (2) to make the ethanol concentration in the solution 25%-95%; further preferably, ethanol is added to make the ethanol concentration in the solution 65%.

[0017] Therapeutic effect of small intestinal mucosa extract on depression:

[0018] Experimental Example 1 The results of the study showed that the small intestinal mucosal extract can significantly improve the depression and anxiety of patients with depression and can significantly improve the learning and memory abilities of patients with depression.

[0019] Experimental Example 2 The results of the study showed that the small intestinal mucosal extract can not only significantly improve the depression and anxiety state of drug-induced depression model rats, but also significantly increase the body weight of depression rats, and its therapeutic effect on depression is significantly better than fluoxetine.

[0020] The present invention also provides a medicine containing the small intestinal mucosa extract. Preferably, the medicine can be a tablet, a granule, a capsule and a syrup. The above medicines can be prepared according to a common preparation process.

[0021] Preferably, the dosage of the above-mentioned drug for treating depression is 0.01 mg / kg-100 mg / kg.

[0022] More preferably, the dosage of the above-mentioned drug for treating depression is 0.2 mg / kg-50 mg / kg.

[0023] The medicine of the present invention may include only active ingredients or may include excipients, antioxidants, pH regulators, preservatives, isotonic agents and any other pharmaceutically and nutritionally acceptable auxiliary materials with guaranteed safety.

[0024] The medicines in the present invention can be prepared into various types, including not only tablets, granules, capsules and syrups, but any other changes to the external form of the product are technical solutions that can be associated with those skilled in the art, so changing the shape of the final product falls within the scope of the present invention.

[0025] Compared with the prior art, the main advantages of the composition provided by the present invention in preventing and treating depression are:

[0026] (1) Significant therapeutic effect and rapid onset of effect.

[0027] (2) Compared with other treatment methods, the main active ingredient in the composition, the small intestinal mucosal extract, is a natural extract, has the advantage of no toxic side effects, and can be prepared into a medicine for oral administration. Specific embodiments

[0028] In order to enable those skilled in the art to fully understand the present invention, the present invention is further described below through specific examples, but those skilled in the art should be aware that the examples of the present invention do not limit the present invention in any way.

[0029] Example 1 Preparation of small intestinal mucosal extract

[0030] (1) Water extraction: Take the pig small intestinal mucosa, homogenize it, add water at a volume-to-mass ratio of 50:1 (ml / g), extract it in a 100°C water bath, and filter the extract to obtain a water extract;

[0031] (2) Alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) to make the ethanol concentration in the solution 85%, and filtering to obtain a precipitate;

[0032] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0033] Example 2 Preparation of small intestinal mucosal extract

[0034] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 10:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0035] (2) Alcohol precipitation: adding 95% ethanol to the aqueous extract obtained in step (1) to make the ethanol concentration in the solution 65%, and filtering to obtain a precipitate;

[0036] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0037] Example 3 Preparation of small intestinal mucosal extract

[0038] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 100:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0039] (2) Alcohol precipitation: adding 25% ethanol to the aqueous extract obtained in step (1) to make the ethanol concentration in the solution 45%, and filtering to obtain a precipitate;

[0040] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0041] Example 4 Preparation of small intestinal mucosal extract

[0042] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 80:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0043] (2) Alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) to make the ethanol concentration in the solution reach 65%, and filtering to obtain a precipitate;

[0044] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0045] Example 5 Preparation of small intestinal mucosal extract

[0046] (1) Extraction: Take the pig small intestinal mucosa, homogenize and crush it, add water at a volume-to-mass ratio of 60:1 (ml / g), perform ultrasonic extraction, and filter the extract to obtain a water extract;

[0047] (2) Alcohol precipitation: adding ethanol to the aqueous extract obtained in step (1) to make the ethanol concentration in the solution 45%, and filtering to obtain a precipitate;

[0048] (3) Freeze-drying: The precipitate obtained in step (3) is freeze-dried to obtain a small intestinal mucosal extract containing mucopolysaccharides and water-soluble proteins.

[0049] Example 6 Preparation of a drug containing a small intestinal mucosal extract

[0050] Take the small intestinal mucosal extract and starch obtained in Example 4, sieve and mix; add starch slurry to make a soft material, sieve, granulate, dry and granulate; then add magnesium stearate, mix and tablet, and obtain tablets.

[0051] Experimental Example 1: Effect of small intestinal mucosal extract on the improvement of depressive behavior in behavioral stress depression model rats

[0052] 1. Test methods

[0053] 80 healthy male SD rats were randomly divided into a normal control group, a model group and an experimental group (Example 4 group, Example 5 group), 20 rats in each group. The model group and the experimental group rats were raised in solitary and chronic mild unpredictable stimulation according to the reference method to construct a depression rat model, and the control group was raised normally. During the modeling period, the experimental group was gavaged with the small intestinal mucosal extract obtained in Example 4 and Example 5 at 20 mg / kg, and the control group and the model group were gavaged with an equal amount of 0.9% NaCl for 4 consecutive weeks. The model group rats were accompanied by typical depression symptoms such as reduced water intake, exploratory behavior, weight loss, decreased interest, and dark fur color.

[0054] 2. Detection indicators

[0055] (1) Open field test

[0056] After the last administration, an open field test was performed according to the reference method. The number of horizontal movements and vertical upright times of the rats within 4 minutes were observed and recorded. The sum of the horizontal and vertical scores was calculated as the number of spontaneous activities, and the immobility time was also recorded.

[0057] (2) Morris water maze test to detect learning and memory function of rats

[0058] After the administration, the rats in each group were subjected to positioning navigation experiment and space exploration experiment using Morris water maze test system according to the reference (ApoE4 age-dependently increases the susceptibility to stress-induced depressive-like behavior and the decline of learning and memory ability).

[0059] 3. Test results

[0060] (1) Effects of small intestinal mucosal extract on depressive behavior in rats with depression

[0061] The number of spontaneous activities of rats in the model group was less than that of the normal control group (P<0.01), while that of the experimental group was more than that of the model group (P<0.01); the immobility time of rats in the model group was longer than that of the normal control group (P<0.01), while that of the experimental group was shorter than that of the model group (P<0.05), indicating that the small intestinal mucosal extract can significantly improve the depression and anxiety state of depression model rats. The specific results are shown in Table 1.

[0062] Table 1 Effects of small intestinal mucosal extract on depressive behavior in rats with depression Note: Compared with the normal control group, ## P<0.01; compared with the model group,& P<0.05, && P<0.01.

[0063] (2) Effects of small intestinal mucosal extract on learning and memory abilities of rats with depression

[0064] The latency of the model group rats in the Morris water maze navigation test on the 1st, 2nd, 3rd, and 4th days was longer than that of the normal control group (P<0.05); while the latency of the experimental group on the 2nd, 3rd, and 4th days was shorter than that of the model group (P<0.05); the number of crossing the platform in the Morris water maze space exploration test in the model group was less than that in the control group (P<0.01), while the number of crossing the platform in the experimental group was more than that in the model group (P<0.05), indicating that the composition can significantly improve the learning and memory ability of depression model rats. The specific results are shown in Table 2.

[0065] Table 2 Effects of small intestinal mucosal extract on learning and memory ability of rats with depression

[0066]

[0067] Note: Compared with the normal control group, # P<0.05, ## P<0.01; compared with the model group, & P<0.05.

[0068] Experimental Example 2 Effect of small intestinal mucosal extract on the depressive behavior of drug-induced depression model rats

[0069] 1. Experimental Methods

[0070] 1.1 Grouping

[0071] Before the experiment, the open-field test was used to perform behavioral scoring. According to the total distance and total number of grids in the open-field test, 60 rats with similar behaviors were randomly divided into 4 groups: blank group, model group, fluoxetine group, and experimental group (Example 4 group and Example 5 group), with 12 rats in each group, and adaptive feeding was performed for 1 week.

[0072] 1.2 Modeling and drug administration

[0073] Referring to the Kulkarni method (Effect of various classes of antidepressants in behavioral paradigms of despair) and improved, the model group and the treatment group (fluoxetine group, experimental group) were given reserpine injection at 0.3 mg / kg intraperitoneal injection, and the blank group was given an equal amount of saline intraperitoneal injection, once a day in the morning for 14 consecutive days. After the modeling was completed, the fluoxetine group was gavaged at 2 mg / kg, and the experimental group was gavaged at 20 mg / kg to give the small intestinal mucosal extract obtained in Example 4 and Example 5. The model group and the blank group were gavaged with an equal amount of distilled water, and the administration cycle was 4 weeks.

[0074] 2. Observation indicators

[0075] 2.1 General

[0076] 2.1.1 Appearance and morphology After each day of the experiment, the number of rats with drooping 1 / 2 of their upper face, hair, morphology (tremor, erect hair, hunched back, curled up), stool status, etc. were observed.

[0077] 2.1.2 Body mass measurement Body mass was measured on the 1st, 7th and 14th day after the formal experiment and before the end of the experiment.

[0078] 2.2 Behavioral science

[0079] 2.2.1 Open field experiment

[0080] The open field experiment was conducted according to the literature. The open field software was debugged before the experiment, and the open field was divided into 4×4 squares. The rats were placed in a 100cm×100cm×40cm open field with four sides painted black. After the experiment started, the camera recorded the movement trajectory of the rats, and the computer recorded the total horizontal movement distance and number of squares of the rats. Each rat was measured for 3 minutes. When conducting the open field experiment, a special lighting lamp for the open field experiment was used, and the experimenter left the open field area to prevent the sound from interfering with the rats. The open field experiment was conducted on the 14th day of the formal experiment.

[0081] 2.2.2 Forced swimming test

[0082] Pour clean water into a bucket (30 cm in diameter, 60 cm in height) to a depth of 40 cm and a water temperature of (25±2)°C. Measure and accumulate the immobility time of the rats within 5 minutes. The rats were slightly curled up but kept in a dangling position. The time when the nostrils were exposed to the water surface was recorded as the immobility time. After the experiment, each rat was dried with a towel and blow-dried. The forced swimming test was conducted on the 14th day of the formal experiment.

[0083] 3 Data processing

[0084] SPSS 20.0 software was used for data processing. One-way analysis of variance was used to compare the sample means among the groups, and the LSD method was used for multiple positive comparisons among the groups; P < 0.05 was considered statistically significant.

[0085] 4 Results

[0086] 4.1 General

[0087] 4.1.1 Appearance and shape

[0088] On the fourth day of the formal experiment, rats in the model group began to experience ptosis of the eyelids, and on the 14th day, 7 rats had ptosis of half of their upper eyelids. On the fifth day, rats in the model group began to experience erect hair, raised arched backs, and loose stools, and all rats experienced the above conditions on the 14th day.

[0089] 4.1.2 Body mass

[0090] There was no statistically significant difference in the baseline weight of rats in each group before modeling (P>0.05). One-way analysis of variance showed that at different measurement time points, compared with the blank group, the body weight of rats in the model group was significantly reduced (P<0.05), and the body weight increment was significantly reduced (P<0.05); compared with the model group, the body weight of the fluoxetine group and the experimental group increased after administration, and the fluoxetine group had a statistical difference with the model group at the 14th day (P<0.05), and the experimental group had a statistical difference with the model group starting from the 7th day (P<0.05). Compared with the model group, the body weight increment of both groups increased significantly (P<0.05), indicating that fluoxetine and the small intestinal mucosal extract of the present invention can increase the body weight of depression model rats, and the small intestinal mucosal extract has a faster onset speed. The specific results are shown in Table 3.

[0091] Table 3 Effects of small intestinal mucosal extract on body weight of rats

[0092]

[0093] Note: Compared with the blank group, # P<0.05; compared with the model group, & P<0.05.

[0094] 4.2 Behavioral

[0095] 4.2.1 Open field test: Compared with the blank group, the total horizontal movement distance and the number of grids of rats in the model group were significantly reduced, which was statistically significant (P<0.01); compared with the model group, the total horizontal movement distance and the number of grids of rats in the fluoxetine group and the experimental group were significantly increased, and the difference was statistically significant (P<0.01). Among them, the increase trend in the experimental group was more significant, and the difference between the two groups was statistically significant (P<0.01). The specific results are shown in Table 4.

[0096] Table 4 Effects of small intestinal mucosal extract on open field behavior in rats

[0097] Note: Compared with the blank group, ## P<0.01; compared with the model group, && P<0.01; compared with the fluoxetine group, $$ P<0.01.

[0098] 4.2.2 Forced swimming test: Compared with the blank group, the immobility time of the rats in the model group was significantly increased, and the difference was statistically significant (P<0.01); compared with the model group, the fluoxetine group and the experimental group could reduce the immobility time of the rats in the forced swimming (P<0.05, P<0.01), and the experimental group reduced the time more significantly than the fluoxetine group, and the difference was statistically significant (P<0.05). The specific results are shown in Table 5.

[0099] Table 5 Effect of small intestinal mucosal extract on immobility time in rats forced swimming

[0100]

[0101] Note: Compared with the blank group, ## P<0.01; compared with the model group, & P<0.05, && P<0.01; compared with the fluoxetine group, $ P<0.05.

[0102] The above results show that small intestinal mucosal extract can significantly improve the depression and anxiety state of depression model rats, and its effect is significantly better than fluoxetine.

Claims

1. Use of a small intestinal mucosal extract in the preparation of a product for preventing and treating depression, characterized in that: The small intestinal mucosa extract is prepared from pig small intestinal mucosa through a water extraction and alcohol precipitation process, and the small intestinal mucosa extract contains mucopolysaccharide and water-soluble protein.

2. The use according to claim 1, characterized in that The small intestinal mucosa extract improves depression and anxiety in depression.

3. The use according to claim 1, characterized in that The small intestinal mucosa extract improves learning and memory abilities in depression.

4. The use according to claim 1, characterized in that The preparation process of the small intestinal mucosa extract comprises the following steps: (1) Extraction: Take pig small intestinal mucosa, add water to extract, filter, and obtain water extract; (2) Alcohol precipitation: adding monohydric alcohol to the aqueous extract obtained in step (1), and filtering to obtain a precipitate; (3) Freeze-drying: freeze-dry the precipitate obtained in step (2).

5. The use according to any one of claims 1 to 3, characterized in that The small intestinal mucosa extract and pharmaceutically acceptable auxiliary materials are prepared into a pharmaceutical preparation.

6. The use according to claim 5, characterized in that The dosage of the pharmaceutical preparation is 0.01 mg / kg-100 mg / kg.

7. The use according to claim 6, characterized in that The dosage of the pharmaceutical preparation is 0.2 mg / kg-50 mg / kg.

8. A small intestinal mucosa extract having a preventive and therapeutic effect on depression, or a medicine containing the small intestinal mucosa extract, characterized in that: The small intestinal mucosa extract is prepared from pig small intestinal mucosa through a water extraction and alcohol precipitation process, and the small intestinal mucosa extract contains mucopolysaccharide and water-soluble protein.