Compound donkey-hide gelatin paste for nourishing liver and clearing heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy and preparation method thereof

Abstract

The present invention provides a compound donkey-hide gelatin paste for nourishing the liver and clearing away heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy, and a preparation method thereof. The present invention extracts effective ingredients in traditional Chinese medicine from mulberry, Poria cocos and ginseng through microbial fermentation, and further treats bergamot with ultrafine grinding technology to improve the functionality and physicochemical properties of bergamot. The compound donkey-hide gelatin paste for nourishing the liver and clearing away heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy is finally prepared by combining Chinese medicines such as Chinese date seeds, honey and donkey-hide gelatin. Compared with traditional Chinese herbal medicine donkey-hide gelatin paste, the donkey-hide gelatin paste prepared by the present invention has a higher content of effective ingredients, more active substances, and a more excellent therapeutic effect, and can also have a certain anti-tumor effect, etc.

Description

Technical Field

[0001] The invention relates to the technical field of donkey-hide gelatin, and in particular to a compound donkey-hide gelatin paste capable of nourishing the liver and clearing away heat, relieving depression and calming the mind, regulating menstruation and assisting pregnancy, and a preparation method thereof. Background Art

[0002] Donkey hide gelatin is a solid gelatin made by boiling and concentrating the dried or fresh skin of the donkey, an equine animal. The name "Donkey hide gelatin" was first recorded in Shennong's Herbal Classic. The raw materials of donkey hide gelatin have undergone the following stages of evolution: mainly cow hide - a mixture of cow hide and donkey hide - mainly donkey hide - donkey hide. It is mild in nature, sweet in taste, and enters the lung, liver, and kidney meridians. It can nourish blood and yin, moisten dryness, and stop bleeding. It is used for blood deficiency and sallow complexion, dizziness and palpitations, muscle weakness, restlessness and insomnia, internal wind movement, dry cough due to lung dryness, hemoptysis due to fatigue, vomiting blood and urine, blood in stool and metrorrhagia, and pregnancy and fetal leakage.

[0003] Mulberries are both medicinal and edible plants with extremely high water content, making them difficult to store and transport for a long time after being picked. Their medicinal value has not yet been effectively developed, and common dried mulberries, mulberry jam, mulberry vinegar, mulberry wine and mulberry tea are all low-end processed products. The deep processing technology of mulberries still mainly relies on microbial fermentation. In addition to producing beverages, wine and vinegar, mulberries can also produce enzymes with specific biologically active ingredients. Enzyme products are usually rich in antioxidant active substances such as phenols and biologically active substances such as vitamins, and have antioxidant, anti-tumor, blood sugar lowering, blood pressure lowering, chronic disease prevention and gastrointestinal regulation functions, which are helpful to the health of the body.

[0004] The seeds of the Chinese jujube seed are the dried seeds of the plant Ziziphus jujuba of the Rhamnaceae family. The main active ingredients include saponins, triterpenoids, flavonoids, etc. It has the effects of calming the mind, improving immunity, nourishing the liver, and sedative hypnosis. It is known as the "Oriental Sleeping Fruit". The extract of Chinese jujube seeds is the first choice for the treatment of insomnia symptoms in clinical Chinese medicine. Summary of the invention

[0005] In view of the deficiencies in the prior art, the purpose of the present invention is to prepare a compound donkey-hide gelatin paste which can nourish the liver and clear away heat, relieve depression and calm the mind, regulate menstruation and aid pregnancy, and a preparation method thereof.

[0006] The first aspect of the present invention provides a compound donkey-hide gelatin paste for nourishing the liver and clearing away heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy. The compound donkey-hide gelatin paste comprises the following raw materials in parts by weight: 36 to 65 parts of traditional Chinese medicine fermentation filtrate, 45 to 55 parts of donkey-hide gelatin, 30 to 60 parts of bergamot, 20 to 35 parts of spinach seeds, 15 to 35 parts of mint, 10 to 20 parts of chrysanthemum, 15 to 45 parts of honey, and 15 to 30 parts of cottonseed sugar; the traditional Chinese medicine fermentation filtrate is obtained by filtering mulberry, ginseng, and poria after fermentation with probiotics.

[0007] Furthermore, the mass ratio of mulberry, ginseng and poria in the traditional Chinese medicine fermentation filtrate is (40-75): (55-90): (35-55).

[0008] The second aspect provides a method for preparing a compound donkey-hide gelatin paste for nourishing the liver and clearing away heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy, the steps of which are as follows:

[0009] (1) Preparation of Chinese medicine fermentation filtrate;

[0010] (2) weighing the various components of the compound donkey-hide gelatin paste according to proportion, and then preparing the bergamot superfine powder;

[0011] (3) Preparation of Ziziphus jujuba seed extract;

[0012] (4) Preparation of chrysanthemum-mint extract;

[0013] (5) After uniformly mixing the Chinese medicine fermentation filtrate obtained in step (1), the bergamot superfine powder obtained in step (2), the jujube seed extract obtained in step (3), donkey-hide gelatin, and the chrysanthemum-mint extract obtained in step (4), honey and cottonseed sugar are added for decoction, and finally a compound donkey-hide gelatin paste is obtained that nourishes the liver and clears away heat, relieves depression and calms the mind, and regulates menstruation and aids pregnancy.

[0014] Furthermore, in the step (1), the preparation method of the Chinese medicine fermentation filtrate is as follows: S1: mulberry, ginseng and poria cocos are respectively steam distilled, the distillate is collected, the drug residue is decocted with water for 3 times, the decoction of mulberry, ginseng and poria cocos is combined with the distillate, and concentrated to obtain a concentrate with a crude drug concentration of 0.5 to 1.2 g / ml; S2: bacterial culture: the seed liquid of Lactobacillus plantarum (CICC21827) stored in a refrigerator at 4°C is taken out and inoculated into an LB medium sterilized by high pressure, 180r / min, 37°C for bacterial activation for 10-12h, the activated bacterial strains were inoculated and then inoculated into another fresh LB medium, and the culture was stopped after shaking culture for 16h under the same conditions to obtain Lactobacillus plantarum (CICC21827) bacterial solution; S3 fermentation culture The Lactobacillus plantarum (CICC21827) bacterial solution in step S2 was centrifuged at 8000r / min and 4°C for 10min, the supernatant was discarded, the bacterial cells were resuspended, and then gradient diluted to 10 5 ~10 6 CFU / ml, the bacterial suspension is inoculated in the concentrated solution sterilized by high pressure steam, and the culture and fermentation are carried out, wherein the strain inoculation volume is 1% to 3% of the volume of the concentrated solution; S4 preparation of Chinese medicine fermentation filtrate: the concentrated solution inoculated with plant lactobacillus is fermented at 35 to 37°C, and the strain concentration in the bacterial suspension reaches (10 9 ~10 10) CFU / ml, the fermentation liquid was allowed to stand for 2-4 hours, the supernatant was centrifuged in a refrigerated centrifuge at 6000-8000 r / min and 4°C for 10 minutes, the supernatant was pasteurized in a water bath at 80°C for 10 minutes, allowed to stand and cool to room temperature, and filtered through a 0.45 μm filter membrane to obtain the mulberry, ginseng and poria fermentation filtrate.

[0015] Furthermore, the preparation method of the bergamot ultrafine powder in the step (2) is: placing the bergamot medicinal material in an oven, drying it at 60° C. to constant weight, using a grinder to coarsely grind it to obtain coarse powder, and ultrafine grinding the coarse powder to obtain bergamot ultrafine powder.

[0016] Furthermore, the preparation of the Chinese jujube seed extract in step (3) is as follows: the slightly fried Chinese jujube seeds are put into water and boiled for 3 times, the decoctions are combined and concentrated to 0.2-0.5 g / ml.

[0017] Furthermore, the specific steps of step (4) are as follows: the preparation of the chrysanthemum-mint extract is as follows: chrysanthemum and mint are mixed in proportion, water is added and boiled for 3 times, the decoctions are combined and concentrated to a chrysanthemum-mint extract with a crude drug concentration of 0.3 to 0.5 g / ml.

[0018] Furthermore, the specific steps of step (5) are as follows: the Chinese medicine fermentation filtrate obtained in step (1), the bergamot superfine powder obtained in step (2), the jujube seed extract obtained in step (3), donkey-hide gelatin, and the chrysanthemum-mint extract obtained in step (4) are mixed, and after mixing evenly, honey and cottonseed sugar are added thereto for decoction, and the medicinal liquid is kept boiling while simmering over low heat until the medicinal liquid is in the shape of "water drops forming beads" to obtain the compound donkey-hide gelatin paste.

[0019] The third aspect provides the use of the above-mentioned donkey-hide gelatin compound paste or the donkey-hide gelatin compound paste prepared by the above-mentioned method in traditional Chinese medicine for nourishing the liver and clearing away heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy.

[0020] Compared with the prior art, the present invention has the following beneficial effects:

[0021] 1. The present invention uses mulberry, ginseng and Poria as raw materials, and ferments them with Lactobacillus plantarum, and produces new active ingredients in the fermented products, such as small molecule peptides and other substances. In addition, the ginsenoside compounds, mulberry active ingredients and Poria active ingredients in the fermented products are all increased by fermentation.

[0022] 2. The preparation of bergamot is ultrafine powder, which can effectively destroy the cell wall of the plant, so that the effective ingredients of bergamot can be fully extracted. In addition, the physical and chemical properties of bergamot ultrafine powder are better than those of coarse powder on sale.

[0023] 3. Raffinose has a prebiotic function, which can promote the colonization of intestinal flora and improve intestinal flora, thereby improving health. In addition, it also produces a synergistic effect with the probiotic fermentation product in the present invention to enhance the drug effect. In addition, the donkey-hide gelatin paste of the present invention also has a certain effect of moistening the intestines and relieving constipation. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 It is a curve diagram showing the effect of each group of samples on the 5-HT level in the mouse hippocampus;

[0025] Figure 2 It is a bar graph showing the effect of each group of samples on mouse tumor mass. DETAILED DESCRIPTION

[0026] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the following embodiments and features in the embodiments can be combined with each other without conflict.

[0027] Example 1

[0028] This embodiment provides a compound donkey-hide gelatin paste for nourishing the liver and clearing away heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy, and the compound donkey-hide gelatin paste comprises the following raw materials in parts by weight: 45 parts of Chinese medicine fermentation filtrate, 50 parts of donkey-hide gelatin, 40 parts of bergamot, 30 parts of spinach seeds, 30 parts of mint, 10 parts of chrysanthemum, 20 parts of honey, and 30 parts of cottonseed sugar. The Chinese medicine fermentation product is obtained by filtration after fermentation of mulberry, ginseng, and Poria with probiotics; wherein the mass ratio of mulberry, ginseng, and Poria in the Chinese medicine fermentation filtrate is 50:70:45.

[0029] Preparation method of compound donkey-hide gelatin paste:

[0030] (1) Preparation of Chinese medicine fermentation filtrate;

[0031] (2) weighing the various components of the compound donkey-hide gelatin paste according to proportion, and then preparing the bergamot superfine powder;

[0032] (3) Preparation of Ziziphus jujuba seed extract;

[0033] (4) Preparation of chrysanthemum-mint extract;

[0034] (5) After uniformly mixing the Chinese medicine fermentation filtrate obtained in step (1), the bergamot superfine powder obtained in step (2), the jujube seed extract obtained in step (3), the donkey-hide gelatin, and the chrysanthemum-mint extract obtained in step (4), honey and cottonseed sugar are added for decoction, and the medicinal liquid is kept boiling while simmering over low heat until the medicinal liquid is in the shape of "water drops forming beads", thereby finally obtaining a compound donkey-hide gelatin paste that nourishes the liver and clears away heat, relieves depression and calms the mind, and regulates menstruation and aids pregnancy.

[0035] In the step (1), the preparation method of the Chinese medicine fermentation filtrate is as follows: S1: mulberry, ginseng and poria cocos are respectively steam distilled, the distillate is collected, the drug residue is decocted with water for 3 times, the decoction of mulberry, ginseng and poria cocos is combined with the distillate, and concentrated to obtain a concentrate with a crude drug concentration of 0.8 g / ml; S2: bacterial culture: the seed liquid of plant lactobacillus (CICC21827) stored in a refrigerator at 4°C is taken out, inoculated into an LB medium sterilized by high pressure, and the bacterial strain is activated at 180 r / min and 37°C for 10 hours, and the activated bacterial strain is inoculated and inoculated into another fresh LB medium, and the culture is stopped after shaking culture under the same conditions for 16 hours; S3: fermentation culture: the activated bacterial strain in step S2 is centrifuged at 8000 r / min and 4°C for 10 minutes, the supernatant is discarded to obtain the bacterial body, the bacterial body is resuspended with physiological saline, and then gradiently diluted to 10 5 CFU / ml, the bacterial suspension was inoculated into the concentrated solution sterilized by high pressure steam for culture and fermentation, wherein the strain inoculation volume was 1% of the volume of the concentrated solution; S4 Preparation of Chinese medicine fermentation filtrate: The concentrated solution inoculated with plant lactobacillus was fermented at 37°C until the strain concentration in the bacterial suspension reached 4.5×10 9 CFU / ml, the fermentation liquid was allowed to stand for 4 hours, the supernatant was centrifuged in a refrigerated centrifuge at 8000r / min and 4°C for 10 minutes, the supernatant was pasteurized in a water bath at 80°C for 10 minutes, allowed to stand and cool to room temperature, and filtered through a 0.45μm filter membrane to obtain the Chinese medicine fermentation filtrate.

[0036] The preparation method of the bergamot ultrafine powder in the step (2) is as follows: placing the bergamot medicinal material in an oven, drying it at 60° C. to constant weight, using a grinder to coarsely grind it to obtain coarse powder, and ultrafine grinding the coarse powder to obtain the bergamot ultrafine powder.

[0037] The preparation of the Chinese jujube seed extract in step (3) is as follows: the slightly fried Chinese jujube seeds are put into water and boiled for 3 times, the decoctions are combined and concentrated to 0.5 g / ml of Chinese jujube seed extract.

[0038] The preparation of the chrysanthemum-mint extract in step (4) is as follows: chrysanthemum and mint are mixed in proportion, water is added and boiled for 3 times, the decoctions are combined and concentrated to a chrysanthemum-mint extract with a crude drug concentration of 0.5 g / ml.

[0039] Example 2

[0040] The preparation method of Example 2 is basically the same as that of Example 1, except that the proportion of the compound donkey-hide gelatin paste is different: the compound donkey-hide gelatin paste is mainly composed of the following raw materials in parts by weight: 55 parts of Chinese medicine fermentation filtrate, 45 parts of donkey-hide gelatin, 30 parts of bergamot, 20 parts of spinach seeds, 15 parts of mint, 17 parts of chrysanthemum, 23 parts of honey, and 20 parts of cottonseed sugar. The Chinese medicine fermentation product is obtained by filtering mulberry, ginseng, and Poria after probiotic fermentation; the mass ratio of mulberry, ginseng, and Poria in the Chinese medicine fermentation filtrate is 40:90:55.

[0041] Comparative Example 1

[0042] The preparation method of Comparative Example 1 is substantially the same as that of Example 1, except that no Chinese medicinal fermentation filtrate is added in step (5).

[0043] Comparative Example 2

[0044] The preparation method of Comparative Example 2 is substantially the same as that of Example 1, except that raffinose is not added in step (5).

[0045] Comparative Example 3

[0046] The preparation method of Comparative Example 3 is substantially the same as that of Example 1, except that the Chinese medicinal fermentation filtrate and raffinose are not added in step (5).

[0047] Comparative Example 4

[0048] The preparation method of Comparative Example 4 is substantially the same as that of Example 1, except that the Lactobacillus plantarum (CICC21827) in step (1) is replaced by Bifidobacterium longum (CICC6191), and both strains are purchased from China Fermented Food Industry Research Institute Co., Ltd.

[0049] Comparative Example 5

[0050] The preparation method of Comparative Example 5 is substantially the same as that of Example 1, except that the Lactobacillus plantarum (CICC21827) in step (1) is replaced by Lactobacillus plantarum (CICC21790), and both strains are purchased from China Fermented Food Industry Research Institute Co., Ltd.

[0051] Comparative Example 6

[0052] The preparation method of Comparative Example 6 is substantially the same as that of Example 1, except that the Chinese date seed extract is not added in step (5).

[0053] Comparative Example 7

[0054] The preparation method of Comparative Example 7 is basically the same as that of Example 1, except that ginseng is not added to the raw material of the traditional Chinese medicine fermentation filtrate prepared in step (1).

[0055] Comparative Example 8

[0056] The preparation method of Comparative Example 8 is substantially the same as that of Example 1, except that the concentrated solution prepared in step (1) S1 is fermented without adding Lactobacillus plantarum (CICC21827).

[0057] Depression relief test

[0058] (1) Sample selection

[0059] Animal samples: SPF grade Kunming mice, female, 8 weeks old, weight 30-35 g, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. Free access to food and water, breeding environment 22-24 ° C, relative humidity 40%-60%.

[0060] Test samples: samples prepared in Examples 1-2 and Comparative Examples 1-8, and escitalopram.

[0061] (2) Modeling

[0062] The experimental animals were randomly divided into 13 groups, with 5 animals in each group. Except for the normal control group, the other 12 groups were modeled. The animals were anesthetized with urethane and fixed. The neck was disinfected with povidone-iodine. A longitudinal incision was made on the midline of the animals. The skin and subcutaneous tissue were bluntly separated, and the bilateral common carotid arteries were separated. The bilateral common carotid arteries were clamped with hemostatic forceps to block blood flow for 5 minutes, and then the hemostatic forceps were released to allow blood to reperfuse for 10 minutes. The bilateral common carotid arteries were clamped with hemostatic forceps again to block blood flow for 5 minutes, and then the hemostatic forceps were released. The incision was sutured and the operation area was disinfected. From the second day after cerebral ischemia, CUMS was performed to establish the PSD model, including: swimming in 4℃ ice water, overnight lighting, water deprivation, food deprivation, tail clamping, moist bedding and cage closing. A total of 7 stimulation methods. One or two stimulations were performed every day for 18 consecutive days.

[0063] (3) Depression relief test

[0064] In addition to the normal control group, the PSD model mice were randomly divided into model control group, escitalopram group (5 mg kg -1 ), Example 1 group (1.5 g·kg -1 ), Example 2 group (1.5 g·kg -1 ), and comparative examples 1-8 (1.5 g·kg -1 The normal control group and the model control group were given blank solvent (0.5% sodium carboxymethyl cellulose), and the other groups were given oral administration once a day for 28 consecutive days, with a dosing volume of 10 mL kg -1 .

[0065] Forced swimming test

[0066] The experiment was conducted on the 20th day. Before the formal experiment, the mice were first acclimatized to the environment in the laboratory for at least 1 hour, and then placed in a forced swimming test apparatus (a transparent cylindrical barrel with a height of 35 cm and a diameter of 15 cm, and a water depth of 35 cm). The mice were allowed to cling to the wall of the tank with their front paws in the water and could not support their bodies with their hind paws. The animals were removed after 15 minutes. During the formal experiment, the animals were placed in the forced swimming test apparatus and the water temperature was maintained at (25±2)℃. They were observed for 6 minutes, and the cumulative immobility time within the last 5 minutes was recorded.

[0067] Detection of 5-HT level in mouse hippocampus

[0068] After the administration, the mice were killed, the brains were removed, the hippocampal tissue was quickly peeled off on ice, the wet weight was measured, and 0.01 mol·L -1 Phosphate buffered saline (PBS) was thoroughly homogenized, and then half of the volume of 6% perchloric acid was added to the homogenate, vortexed thoroughly, and spun at 16000 r / min at 4°C. -1 Centrifuge for 20 minutes. Take 100 μL of the supernatant, filter it through a 0.45 μm filter membrane, and then detect it by high performance liquid chromatography.

[0069] (4) Test results

[0070] Results of the forced swimming test

[0071] Table 1 Swimming immobility time of mice in each group

[0072] Group Mouse swimming immobility time (t / s) Normal control group 130±4 Model control group 233±3 Example 1 159±6 Example 2 155±5 Comparative Example 1 208±4 Comparative Example 2 168±6 Comparative Example 3 200±7 Comparative Example 4 209±6 Comparative Example 5 203±4 Comparative Example 6 213±9 Comparative Example 7 209±5 Comparative Example 8 214±6 Escitalopram group 165±3

[0073] Table 1 is a comparison of the immobility time of mice in each group. As can be seen from Table 1, the immobility time of the normal group mice is 130s, while the time of the model group is 233s, which shows that the modeling of the experimental mice is successful, and the mice after modeling show a state of depression, so the immobility time is much longer than that of the normal group. However, after drug treatment, the immobility time of the mice in Example 1-2 groups was reduced to about 155s, while the time of the escitalopram group was 165s, which shows that the samples prepared in Examples 1-2 of the present invention can treat depression. And from the comparison of the data of Example 1 and Comparative Example 2, it can be seen that the immobility time of the mice in Example 1 is lower than that of the mice in Comparative Example 2, but the difference is not significant, indicating that the addition of cottonseed sugar has little effect on the treatment of depression. Ziziphus jujuba seed has a tranquilizing and anti-depressant effect, but without adding the traditional Chinese medicine fermentation filtrate (Comparative Example 1), the swimming immobility time of the mice in Comparative Example 1 was significantly increased compared with that in Example 1, and when Ziziphus jujuba seed was not added in Comparative Example 6, the swimming immobility time of the mice in Comparative Example 6 was equivalent to that in Comparative Example 1, which shows that the Ziziphus jujuba seed extract can play a synergistic role with the fermentation filtrate, making the tranquilizing and anti-depressant effect of the sample more excellent.

[0074] Detection of 5-HT level in mouse hippocampus

[0075] Figure 1 The curve diagram of the detection of 5-HT level in the hippocampus of mice of each sample. It can be seen from the figure that compared with the normal control group, the 5-HT level in the hippocampus of mice in the model control group was significantly reduced. The 5-HT level in the hippocampus of mice treated with escitalopram was significantly increased. The 5-HT level in the hippocampus of Examples 1 to 2 was also significantly increased, which shows that the samples of Examples 1 to 2 can affect the level of monoamine neurotransmitters, thereby affecting the depressive behavior of mice.

[0076] Menstrual regulation test

[0077] (1) Sample selection

[0078] Animal samples: SPF grade Kunming mice, female, 10 weeks old, weight 40-45 g, purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd. Free access to food and water, breeding environment 22-24 ° C, relative humidity 40%-60%.

[0079] Test samples: samples prepared in Examples 1-2 and Comparative Examples 1-8.

[0080] (2) Modeling

[0081] The experimental animals were randomly divided into 12 groups, 5 in each group. Except for the normal control group, the other 11 groups were modeled, and each animal was subcutaneously injected with human chorionic gonadotropin twice a day for 3 weeks; insulin was gradually increased from 0.5UPd to 6.0UPd from the 1st to the 11th day, and maintained at this dose until the 21st day. The normal control group was subcutaneously injected with normal saline every day for 3 weeks.

[0082] (3) Method of administration

[0083] After the mouse model was successfully established, the mice were treated with drugs. Example 1-2 The samples prepared in Comparative Examples 1-8 were dissolved and then gavage was performed on the corresponding mice. The dosage was 0.1 g / 10 g, and the gavage dosage was 0.1 mL·10 g. -1 The model group and the blank control group were given 0.1 mL·10 g -1 The mice were gavaged with distilled water once a day for 21 consecutive days.

[0084] (4) Detection method

[0085] After the administration, blood was collected from the retro-orbital vein of the mice in the fasting state, and the blood serum was separated by centrifugation. The FSH and LH contents were determined by radioimmunoassay.

[0086] (5) Test results

[0087] Table 2 Effects of each group of samples on the levels of FSH and LH in rat serum

[0088]

[0089]

[0090] PCOS is an endocrine disorder syndrome caused by hypothalamus-pituitary-ovarian dysfunction. High LH is the basic pathophysiological characteristic of endocrine disorders. Compared with the blank control group, the content of FSH and LH in the model control group was significantly increased, indicating that the modeling of this experiment was successful. Compared with the model control group, the content of LH and FSH in Example 1-2 groups was significantly reduced, indicating that the examples prepared by the present invention can regulate gynecological endocrine disorders. The data of Comparative Examples 4-5, 7-8 are compared with the data of Example 1. It can be seen that after the mulberry, Poria cocos and ginseng in the traditional Chinese medicine fermentation filtrate prepared by the present invention are fermented by Lactobacillus plantarum (CICC21827), the components work synergistically, so that the endocrine regulation effect of the compound donkey-hide gelatin prepared in Example 1 is more excellent. From the data comparison between other comparative examples and Example 1, it can be seen that the Chinese medicine fermentation filtrate in the compound donkey-hide gelatin paste has a synergistic effect with raffinose. The reason may be that raffinose can promote the active ingredients in the fermentation liquid to take effect, and raffinose has certain prebiotic functions. Therefore, the prepared example samples have excellent effects on regulating menstruation and aiding pregnancy.

[0091] Anti-tumor effect detection

[0092] (1) Sample selection

[0093] Animal samples: 55 SPF female Kunming grade mice were purchased from Beijing Weitonglihua Experimental Animal Technology Co., Ltd.

[0094] Test sample: Compound donkey-hide gelatin paste prepared in Examples 1-2 and Comparative Examples 1-8

[0095] (2) Cell culture

[0096] CT26 cells were cultured in 1640 medium supplemented with 10% fetal bovine serum and 1% penicillin / streptomycin double antibody solution and incubated in a constant temperature incubator at 37°C with 5% CO2.

[0097] (3) Construction of mouse orthotopic rectal tumor model

[0098] CT26 cells were cultured in a conventional manner. When the tumor was inoculated, mice were anesthetized by intraperitoneal injection of 2% sodium pentobarbital, with each mouse injected with 80 μl. After anesthesia, 500,000 cells / mouse were injected into the submucosal layer of the mouse rectum through a sterile syringe, and the tumor growth was then observed.

[0099] (4) Anti-tumor effect detection

[0100] The 55 mice with successful modeling were randomly divided into 11 groups, 5 mice in each group. They were named as: model control group, embodiment 1 group, embodiment 2 group, comparative example 1 group, comparative example 2 group, comparative example 3 group, comparative example 4 group, comparative example 5 group, comparative example 6 group, comparative example 7 group, comparative example 8 group. Treatment was started on the 14th day after successful modeling, and the time point of starting treatment was recorded as day 0, i.e. D0. The corresponding group samples were used for treatment at D0, D7, and D14, respectively, with a dosage of 0.25g / each time, and the control group used the same dose of sterile PBS for control treatment. The mice were euthanized on D21 day. The weight of the mice was detected every three days during the experiment to observe the health status of the mice.

[0101] (5) Test results

[0102] The tumor mass of each group was weighed. Figure 2 The result data.

[0103] like Figure 2 As shown, the average tumor mass of Example 1 group was 1.8 g, while the average tumor mass of Comparative Example 1 group was 4.6 g, the average tumor mass of Comparative Example 2 group was 3.8 g, the average tumor mass of Comparative Example 3 group was 5.5 g, the average tumor mass of Comparative Example 4 and Comparative Example 5 groups were 4.4 g and 4.3 g, respectively, the average tumor mass of Comparative Example 6 group was 2.9 g, the average tumor mass of Comparative Example 7 group was 5.3 g, and the average tumor mass of Comparative Example 8 group was 5.6 g.

[0104] From the above data comparison, it can be seen that the samples of Examples 1 to 2 prepared by the present invention have obvious anti-tumor effects. The reason for this is that after the Chinese medicine fermentation filtrate of the compound donkey-hide gelatin paste of Example 1 is fermented by plant lactobacillus (CICC21827), the Chinese medicine fermentation filtrate produced has a certain antibiotic effect and can play an anti-tumor effect. In addition, there is a synergistic effect between the fermentation filtrate and other components in the medicine, making the anti-tumor effect of the embodiment group more excellent. Compared with Comparative Examples 1-3, the average tumor mass of Example 1 is significantly reduced, indicating that the Chinese medicine fermentation filtrate after cottonseed sugar and mulberry, Poria cocos, and ginseng are fermented by specific plant lactobacillus (CICC21827) can play a synergistic role, making the anti-tumor effect of the embodiment sample more excellent.

Claims

1. A compound donkey-hide gelatin paste for relieving depression, calming the nerves, regulating menstruation and aiding pregnancy, characterized in that: The compound donkey-hide gelatin paste comprises the following raw materials in parts by weight: 36-65 parts of Chinese medicine fermentation filtrate, 45-55 parts of donkey-hide gelatin, 30-60 parts of bergamot, 20-35 parts of spinach seeds, 15-35 parts of mint, 10-20 parts of chrysanthemum, 15-45 parts of honey, and 15-30 parts of cottonseed sugar; the Chinese medicine fermentation filtrate is obtained by filtering mulberry, ginseng, and poria after probiotic fermentation; The mass ratio of mulberry, ginseng and poria in the traditional Chinese medicine fermentation filtrate is (40-75): (55-90): (35-55); The preparation method of the compound donkey-hide gelatin paste is: (1) Preparation of Chinese medicine fermentation filtrate; (2) Weighing the various components of the compound donkey-hide gelatin paste according to proportion, and then preparing the bergamot ultrafine powder; (3) Preparation of Ziziphus jujuba seed extract; (4) Preparation of chrysanthemum-mint extract; (5) after uniformly mixing the Chinese medicine fermentation filtrate obtained in step (1), the bergamot superfine powder obtained in step (2), the jujube seed extract obtained in step (3), donkey-hide gelatin, and the chrysanthemum-mint extract obtained in step (4), adding honey and cottonseed sugar for decoction, finally obtaining a compound donkey-hide gelatin paste for nourishing the liver, clearing away heat, relieving depression and calming the mind, regulating menstruation and aiding pregnancy; In the step (1), the preparation method of the Chinese medicine fermentation filtrate is as follows: S1, mulberry, ginseng, and poria are respectively distilled by steam, the distillate is collected, the drug residue is decocted with water for 3 times, the decoction of mulberry, ginseng, and poria is combined with the distillate, and concentrated to obtain a concentrate with a crude drug concentration of 0.5-1.2 g / ml; S2, bacterial culture: the seed liquid of Lactobacillus plantarum CICC21827 stored in a refrigerator at 4°C is taken out and inoculated into an LB medium sterilized by high pressure for 18 0r / min, 37°C for strain activation for 10-12h, the activated strain was inoculated into another fresh LB medium, and the culture was stopped after shaking culture for 16h under the same conditions to obtain Lactobacillus plantarum CICC21827 bacterial solution; S3, fermentation culture: the Lactobacillus plantarum CICC21827 bacterial solution in step S2 was centrifuged at 8000r / min and 4°C for 10min, the supernatant was discarded to take the bacterial cells, the bacterial cells were resuspended, and then gradient diluted to 10 5 ~10 6 CFU / ml, the bacterial suspension was inoculated into the concentrated solution sterilized by high pressure steam for culture and fermentation, wherein the strain inoculation volume was 1%~3% of the volume of the concentrated solution; S4, preparation of Chinese medicine fermentation filtrate: the concentrated solution inoculated with plant lactobacillus was fermented at 35~37℃, and the strain concentration in the bacterial suspension reached (10 9 ~10 10 ) CFU / ml to obtain fermentation liquid, let the fermentation liquid stand for 2-4 hours, take the supernatant and centrifuge it in a refrigerated centrifuge at 6000-8000r / min and 4℃ for 10 minutes, take the supernatant and pasteurize it in a water bath at 80℃ for 10 minutes, let it stand and cool to room temperature, and filter it with a 0.45μm filter membrane to obtain the traditional Chinese medicine fermentation filtrate; Preparation of the Chinese jujube seed extract in step (3): Put the slightly fried Chinese jujube seeds into water and decoct them three times, combine the decoctions, and concentrate them to obtain a Chinese jujube seed extract with a crude drug concentration of 0.2-0.5 g / ml; The preparation of the chrysanthemum-mint extract in step (4) is as follows: chrysanthemum and mint are mixed in proportion, water is added and boiled for 3 times, the decoctions are combined and concentrated to a chrysanthemum-mint extract with a crude drug concentration of 0.3-0.5 g / ml.

2. The compound donkey-hide gelatin paste for relieving depression, calming the nerves, regulating menstruation and aiding pregnancy according to claim 1, characterized in that: The preparation method of the bergamot ultrafine powder in step (2) is as follows: placing the bergamot medicinal material in an oven, drying it at 60° C. to constant weight, using a grinder to coarsely grind it to obtain coarse powder, and ultrafine grinding the coarse powder to obtain the bergamot ultrafine powder.

3. The compound donkey-hide gelatin paste for relieving depression, calming the nerves, regulating menstruation and aiding pregnancy according to claim 2, characterized in that: The specific steps of step (5) are as follows: the Chinese medicine fermentation filtrate obtained in step (1), the bergamot superfine powder obtained in step (2), the jujube seed extract obtained in step (3), donkey-hide gelatin, and the chrysanthemum-mint extract obtained in step (4) are mixed, and after the mixture is evenly mixed, honey and cottonseed sugar are added thereto for decoction, and the medicinal liquid is kept boiling while being simmered over low heat until the medicinal liquid is in the shape of "water drops forming beads" to obtain the compound donkey-hide gelatin paste.

4. Use of the donkey-hide gelatin compound paste according to any one of claims 1 to 3 in the preparation of a traditional Chinese medicine for relieving depression, calming the nerves, regulating menstruation and aiding pregnancy.

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