A combined ferment of ginseng, astragalus membranaceus and donkey-hide gelatin, its preparation method and uses

Abstract

The present invention provides a combined ferment of ginseng, astragalus membranaceus and donkey-hide gelatin, its preparation method and uses, relating to the technical field of pharmaceutical preparations. The combined ferment is obtained by fermenting ginseng, astragalus membranaceus and donkey-hide gelatin, and the mass ratio of ginseng, astragalus membranaceus and donkey-hide gelatin is 1-3:10-30:1. By performing probiotic fermentation on the composition of ginseng, astragalus membranaceus and donkey-hide gelatin, the present invention converts macromolecular saponins into sapogenins or smaller-molecule secondary saponins, cleaves proteins into smaller-molecule peptide chains, and cleaves polysaccharides into smaller-molecule sugar chains, which is beneficial to the absorption and utilization of the effective components of the drug and can quickly play the role of supplementing both qi and blood.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and particularly relates to a combined ferment of ginseng, astragalus and donkey-hide gelatin, and its preparation method and uses. Background Art

[0002] Ginseng is the dried root and rhizome of the Araliaceae plant Panax ginseng C.A.Meyer. It tastes sweet, slightly bitter, and slightly warm, and has the functions of greatly tonifying primordial qi, restoring pulse and arresting collapse, invigorating the spleen and replenishing the lung, promoting the production of body fluid and nourishing blood, calming the mind and improving intelligence. Ginseng greatly tonifies primordial qi, with the emphasis on rescuing from collapse and tonifying the internal deficiency of the body. The ginsenosides and polysaccharides contained in it are the main active ingredients. Astragalus is the dried root of the Leguminosae plant Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge.) Hsiao or Astragalus membranaceus (Fisch.) Bge. It tastes sweet and warm, and has the effects of invigorating qi and lifting yang, benefiting the defensive qi and strengthening the superficies, inducing diuresis and alleviating edema, and expelling toxins and promoting granulation. It mainly acts on the nutrient qi and defensive qi, especially on tonifying the defensive qi, that is, the deficiency of the superficies of the body. It mainly contains astragalosides, astragalus polysaccharides, etc. Donkey-hide gelatin is made by decocting and concentrating the skin of the Equidae animal Equus asinus L. It tastes sweet and neutral, and has the effects of enriching blood and stopping bleeding, nourishing yin and moistening dryness, and preventing miscarriage. 80% of the components in donkey-hide gelatin are collagen, amino acids and peptides, but it has the drawback of "hindering the stomach due to its greasy nature". Ginseng is partial to yin and tonifies the middle-jiao, while astragalus is partial to yang and strengthens the superficies. The combination of the two herbs, one yin and one yang, interact with each other and jointly play the role of strengthening the healthy qi and supplementing qi. "Ben Cao Meng Quan" points out that ginseng regulates the middle-jiao, and astragalus strengthens the superficies, achieving the effect of supplementing qi on the exterior and interior. Together with donkey-hide gelatin for enriching blood and nourishing yin, moistening the lung and stopping bleeding, all three belong to the herbs for strengthening the healthy qi, taking into account qi, blood, yin, yang and zang-fu organs, achieving the effect of taking into account both the exterior and interior and supplementing both qi and blood.

[0003] Traditional Chinese medicine believes that qi is the commander of blood, and blood is the mother of qi. The clinical manifestations of qi and blood deficiency syndrome include pale or sallow complexion, dizziness, vertigo, shortness of breath and reluctance to speak, fatigue, chest tightness, insomnia and other symptoms. In recent years, qi and blood deficiency syndrome is a common disease in clinical practice. Qi and blood deficiency syndrome can be seen in chemotherapy, radiotherapy, surgery, senility, postpartum, chronic illness, etc., with symptoms such as physical fatigue, low immunity, weak spleen and stomach, bone marrow suppression, reduction of red blood cells and white blood cells. And cardiovascular and cerebrovascular diseases, hypertension, coronary heart disease, heart failure, chronic obstructive pulmonary disease, diabetes, etc. will all be accompanied by the occurrence of qi deficiency syndrome. At present, the traditional Chinese medicine compound prescriptions for treating qi and blood weakness have many medicinal flavors in the prescription, large dosages, slow onset of action, and most of the effective components of the compound are macromolecular substances, with low bioavailability and low intestinal absorption, which will lead to a longer treatment cycle and the treatment obstacle of "the weak cannot be tonified".

[0004] Patients with a weak constitution often have a disordered intestinal flora and weakened digestion and absorption abilities. For example, the hydrolysis of donkey-hide gelatin collagen requires a large amount of proteases. However, patients with qi and blood deficiency have insufficient digestive enzymes due to the disordered intestinal flora, resulting in a decline in digestive ability. Therefore, after taking donkey-hide gelatin, not only does the digestive burden increase, but it also affects the absorption of the effective components of donkey-hide gelatin.

[0005] Probiotics are a type of microorganisms beneficial to the host and are widely present in the human intestinal tract and reproductive system, playing an important role in the digestion and absorption of food and drugs. Using probiotics to transform saponins, polysaccharides, and peptide components in traditional Chinese medicine can promote the conversion of effective components into smaller molecular substances that are more easily absorbed by the human body, and even produce new substances with stronger activity.

[0006] For example, Chinese Patent CN112618691A discloses a traditional Chinese medicine composition with the efficacy of supplementing qi and blood, its preparation method and application. The preparation is as follows: S1: After cleaning and removing impurities from wild jujube seeds, lily, polygonatum odoratum, ginseng, carob, and Chinese dates, they are pulverized and water is added and stirred evenly to obtain a mixed solution; a culture medium containing Saccharomyces boulardii, Lactobacillus plantarum, and Lactobacillus bulgaricus is added for mixed fermentation. After the fermentation ends, a concentrated solution is obtained through filtration and reduced-pressure concentration steps; S2: After cleaning and removing impurities from hawthorn, Rosa rugosa cv. Plena, wolfberry, fried germinated barley, and dried ginger, they are extracted, and a concentrated solution is obtained through a low-temperature reduced-pressure concentration step; S3: The above concentrated solutions are combined, excipients such as maltitol are added, and through the steps of sugar boiling and paste making, and then packaged to obtain the product. The process of this invention can improve the extraction rate of the effective components of traditional Chinese medicine, protect the effective components in traditional Chinese medicinal materials from being destroyed, degrade macromolecular substances to produce new active components such as polysaccharides and ginsenosides, and at the same time reduce the toxic and side effects of traditional Chinese medicine itself. Summary of the Invention

[0007] Based on the prior art, the present invention provides a combined ferment of ginseng, astragalus membranaceus, and donkey-hide gelatin, its preparation method and uses. By performing probiotic fermentation on the composition of ginseng, astragalus membranaceus, and donkey-hide gelatin, macromolecular saponins are converted into aglycones or smaller molecular secondary saponins, proteins are cleaved into smaller molecular peptide chains, and polysaccharides are cleaved into smaller molecular sugar chains, which is beneficial to the absorption of the drug and can quickly exert the therapeutic effect of supplementing both qi and blood.

[0008] In order to achieve the above object, the present invention is implemented by adopting the following technical solutions:

[0009] On the one hand, the present invention provides a combined ferment of ginseng, astragalus membranaceus, and donkey-hide gelatin, and the combined ferment is obtained by fermenting ginseng, astragalus membranaceus, and donkey-hide gelatin, and the mass ratio of ginseng, astragalus membranaceus, and donkey-hide gelatin is 1 - 3:10 - 30:1.

[0010] The fermentation is carried out using probiotics; the probiotics are one or several of lactobacillus and / or bifidobacterium;

[0011] The Lactobacillus mentioned above is selected from Lactobacillus bulgaricus and / or Lactobacillus plantarum.

[0012] Preferably, the probiotic is a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum, and the mass ratio of the two is 1:1.

[0013] As some preferred embodiments, the combined ferment is obtained by fermenting 1-3 parts of ginseng, 10-30 parts of astragalus membranaceus, and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0014] As a preferred embodiment, the combined ferment is obtained by fermenting 2 parts of ginseng, 25 parts of astragalus membranaceus, and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0015] Ginseng mainly contains active ingredients such as ginsenosides and polysaccharides, greatly tonifying primordial qi and focusing on tonifying the deficiency inside the body. Astragalus membranaceus mainly tonifies the deficiency on the body surface and mainly contains astragalosides and astragalus polysaccharides. Donkey-hide gelatin nourishes blood and mainly contains collagen, amino acids, and peptides.

[0016] For the combined ferment provided by the present invention, donkey-hide gelatin is "greasy and obstructive to the stomach", and the components contained in ginseng and astragalus membranaceus are all ginsenosides, astragalosides, polysaccharides, etc. with relatively large molecular weights, which are difficult to digest and absorb, and the bioavailability is relatively low. For the weak group with mostly disordered intestinal flora, "the deficiency is not tonifiable", and a good tonifying effect cannot be achieved. Therefore, it is necessary to carry out in vitro fermentation and degradation with probiotics to produce secondary low-molecular ginsenosides, astragalosides, and small-molecular polysaccharides and other components that are more easily absorbed and have stronger activity.

[0017] The chemical components contained in the combined ferment are fermented and degraded by probiotics, producing rare ginsenosides, secondary astragalosides, secondary flavonoid glycosides, oligopeptides, amino acids, small-molecular polysaccharides, etc. that are more easily absorbed, have stronger activity, and have smaller molecular weights. Probiotics ferment the composition of ginseng, astragalus membranaceus, and donkey-hide gelatin in vitro, realizing the prior fermentation and degradation of the chemical components contained in this traditional Chinese medicine composition by probiotics, and reducing the digestive burden of patients with physical weakness. Fermentation with probiotics can significantly increase the content of rare ginsenosides Rh1, Rc, and Rg3(S), and even produce some new active secondary metabolites.

[0018] The present invention also provides a preparation method of the above-mentioned combined ferment, including the following steps:

[0019] (1) Weigh the formula dosage of ginseng and astragalus membranaceus, reflux and extract with water, combine the filtrates, concentrate, and set aside;

[0020] (2) Crush donkey-hide gelatin, add it to the filtrate in step (1) to dissolve, and then let it cool to obtain a mixed solution for standby;

[0021] (3) Add the probiotic powder into sterile physiological saline respectively to prepare a bacterial suspension, add the bacterial suspension into the mixture in step (2), conduct fermentation culture, and sterilize to obtain the combined fermented product as described above.

[0022] In the reflux extraction described in step (1) above, the number of times is 2 - 3 times, the time for each time is 1 - 2 h, and the water addition amount is 2 - 5 times the total mass of ginseng and astragalus; preferably 4 times.

[0023] In step (2) above, the temperature for cooling is 36 - 38 °C.

[0024] In step (3) above, the temperature for fermentation is 36 - 39 °C, and the time is 36 - 48 h.

[0025] In step (3) above, the concentration of the bacterial suspension is 1 - 2 mg / mL; preferably 1 mg / mL.

[0026] In step (3) above, the mass of the bacterial suspension is 1% - 2% of the total mass of ginseng, astragalus and donkey-hide gelatin; preferably 1%.

[0027] The present invention also provides the application of the above combined fermented product in the preparation of a medicament for treating or preventing qi and blood deficiency syndrome.

[0028] The qi and blood deficiency syndrome described above includes qi and blood deficiency, physical fatigue, low immunity or weak spleen and stomach caused by chemotherapy, radiotherapy, surgery, old age and debility, postpartum or long-term illness.

[0029] The present invention also provides the application of the above combined fermented product in the preparation of a medicament for treating or preventing leukopenia.

[0030] The present invention also provides the application of the above combined fermented product in the preparation of a medicament or health product for improving sleep.

[0031] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0032] Based on the etiology of intestinal flora disorder, weakened digestion and absorption ability in patients with physical weakness, the combined fermented product provided by the present invention solves the treatment obstacle of "weakness not tolerating supplementation" in traditional Chinese medicine clinical practice, and realizes the treatment purpose of quickly supplementing qi and blood. It has the effects of replenishing qi, nourishing blood and promoting blood circulation; it is used as an auxiliary tonic for patients with qi and blood deficiency syndrome and tumor patients after radiotherapy and chemotherapy, can relieve the symptoms of qi and blood deficiency, improve the body immunity, improve the blood picture indexes such as red blood cell RBC, white blood cell WBC, hemoglobin HGB, platelet count PLT, etc., is quickly absorbed, can be directly drunk, and has no toxic and side effects. Detailed implementation manners

[0033] Next, the technical solutions in the embodiments of the present invention will be clearly and completely described in conjunction with the tables in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0034] Next, the embodiments and comparative examples of the present invention are shown to more specifically illustrate the composition of the present invention, but the present invention is not limited thereto.

[0035] Example 1 A combined ferment of ginseng, astragalus membranaceus and donkey-hide gelatin and its preparation method

[0036] The combined ferment is obtained by fermenting 1 part of ginseng, 10 parts of astragalus membranaceus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0037] The specific preparation method is as follows:

[0038] (1) Weigh the formula amount of ginseng and astragalus membranaceus, add 2 times the amount of water and reflux extract for 2 times, 2 hours each time, combine the filtrates, concentrate, and reserve;

[0039] (2) Crush the formula amount of donkey-hide gelatin, add it to the filtrate of step (1) to dissolve, and then cool to 36 °C to obtain a mixed solution for reserve;

[0040] (3) Add the Lactobacillus bulgaricus powder and Lactobacillus plantarum powder to sterile physiological saline respectively to prepare a bacterial suspension of 1 mg / mL, add the bacterial suspension to the mixed solution of step (2), the volume of the bacterial suspension is 1% of the volume of the mixed solution, carry out fermentation culture, the fermentation temperature is 36 °C, the time is 48 hours, and sterilize to obtain the combined ferment of the composition.

[0041] Example 2 A combined ferment of ginseng, astragalus membranaceus and donkey-hide gelatin and its preparation method

[0042] The combined ferment is obtained by fermenting 2 parts of ginseng, 30 parts of astragalus membranaceus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0043] The specific preparation method is as follows:

[0044] (1) Weigh the formula amount of ginseng and astragalus membranaceus, add 5 times the amount of water and reflux extract for 3 times, 1 hour each time, combine the filtrates, concentrate, and reserve;

[0045] (2) Crush the formula amount of donkey-hide gelatin, add it to the filtrate of step (1) to dissolve, and then cool to 38 °C to obtain a mixed solution for reserve;

[0046] (3) Add the powder of Lactobacillus bulgaricus and the powder of Lactobacillus plantarum into sterile physiological saline respectively to prepare a bacterial suspension of 2 mg / mL. Add the bacterial suspension into the mixture in step (2), and the volume of the bacterial suspension is 1% of the volume of the mixture. Then carry out fermentation culture at a fermentation temperature of 39 °C for 36 hours, and sterilize to obtain the fermented product of the composition.

[0047] Example 3 A combined fermented product of ginseng, astragalus membranaceus and donkey-hide gelatin and its preparation method

[0048] The combined fermented product is obtained by fermenting 2 parts of ginseng, 25 parts of astragalus membranaceus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum at a mass ratio of 1:1.

[0049] The specific preparation method is as follows:

[0050] (1) Weigh the ginseng and astragalus membranaceus in the formula amount, add 4 times the amount of water and reflux extract for 3 times, 1.5 hours each time. Combine the filtrates and concentrate for standby.

[0051] (2) Crush the donkey-hide gelatin in the formula amount, add it to the filtrate in step (1) to dissolve, and then cool it to 37 °C to obtain a mixture for standby.

[0052] (3) Add the powder of Lactobacillus bulgaricus and the powder of Lactobacillus plantarum into sterile physiological saline respectively to prepare a bacterial suspension of 1 mg / mL. Add the bacterial suspension into the mixture in step (2), and the volume of the bacterial suspension is 1% of the volume of the mixture. Then carry out fermentation culture at a fermentation temperature of 38 °C for 40 hours, and sterilize to obtain the fermented product of the composition.

[0053] Comparative Example 1

[0054] The difference from Example 3 is that only Lactobacillus bulgaricus is used. Others are the same as in Example 3.

[0055] Comparative Example 2

[0056] The difference from Example 3 is that only Lactobacillus plantarum is used. Others are the same as in Example 3.

[0057] Comparative Example 3

[0058] The difference from Example 3 is that no fermentation treatment is carried out. Others are the same as in Example 3.

[0059] Comparative Example 4

[0060] The difference from Example 3 is that astragalus membranaceus is replaced by codonopsis pilosula. Others are the same as in Example 3.

[0061] Comparative Example 5

[0062] The difference from Example 3 is that ginseng is replaced by Codonopsis pilosula. Others are the same as Example 3.

[0063] Effect experiment:

[0064] 1. Detection of the contents of rare ginsenosides Rh1, Rc, and Rg3(S) in the combined fermentate

[0065] Method: The HPLC method was used, with a UV detector, a reversed-phase chromatographic column Agilent ZORBAX SB-C18 4.6×250 mm, 5 μm; selected, the detection wavelength was 203 nm, the column temperature was 30 °C, the flow rate was 1 ml / min, acetonitrile was used as mobile phase A, and 0.05% (v / v) phosphoric acid water was used as mobile phase B for gradient elution. The gradient elution conditions were as follows: 0 - 14 min, 20 - 21% A; 14 - 24 min, 21 - 22% A; 24 - 36 min, 22 - 23% A; 36 - 52 min, 23 - 25% A; 52 - 55 min, 25 - 28% A; 55 - 78 min, 28 - 30% A; 78 - 96 min, 30 - 35% A; 96 - 115 min, 35 - 60% A.

[0066] Reference substances: Ginsenosides Rh1, Rc, and Rg3(S), purchased from Chengdu Mansite Biotechnology Co., Ltd.

[0067] The detection results are shown in Table 1 below.

[0068] Table 1

[0069]

[0070] As can be seen from Table 1 above, under the action of different probiotic compositions, the contents of ginsenosides Rh1, Rc, and Rg3(S) increased to varying degrees. Among them, the contents of ginsenosides Rh1, Rc, and Rg3(S) in the fermented composition prepared in Example 3 were the highest; in each comparative example, after single-strain fermentation, the contents of ginsenosides Rh1, Rc, and Rg3(S) increased compared with the unfermented group, but were significantly lower than those in the example group. This shows that fermentation treatment is beneficial to the dissolution of ginsenosides Rh1, Rc, and Rg3(S).

[0071] 2. Detection of the content of cycloastragenol in the combined fermentate

[0072] Method: The HPLC-DAD-ELSD method was adopted, with a UV detector, a reverse-phase chromatographic column Ultimate XB-C18 4.6×250 mm 5 μm; selected, the detection wavelength was 260 nm, the carrier gas flow rate: 3.2 L / min, the drift tube temperature: 110 °C, the gain: 1, the flow rate: 1 ml / min. Acetonitrile was used as mobile phase A, and 0.1% (v / v) formic acid in water was used as mobile phase B for gradient elution. The gradient elution conditions were as follows: 0 - 10 min, 18% A; 10 - 32 min, 18 - 35% A; 32 - 40 min, 35% A; 40 - 57 min, 35 - 53% A; 57 - 67 min, 53 - 68% A.

[0073] Reference substance: Cycloastragenol, purchased from Chengdu Mansite Biotechnology Co., Ltd.

[0074] The detection results are shown in Table 2 below.

[0075] Table 2

[0076]

[0077] As can be seen from Table 2 above, under the action of different probiotic compositions, the content of cycloastragenol, the aglycone of astragaloside IV in astragalus, increased to varying degrees. Among them, the content in Example 3 was the highest, reaching 2.8715 mg; in each comparative example, after single-strain fermentation, the content of cycloastragenol increased compared with the non-fermented group, but was significantly lower than that of the example group. It indicates that astragaloside IV may be enzymatically hydrolyzed under the action of probiotics during fermentation, resulting in an increase in the content of the aglycone.

[0078] 3. Effect of the combined fermentation product on the increase in white blood cells in a rat model of cyclophosphamide-induced myelosuppression

[0079] Experimental animals: 130 male SD rats, weighing 220 ± 20 g.

[0080] Drugs: The combined fermentation products prepared in Examples 1 - 3 and Comparative Examples 1 - 5, positive drug: Baoyuan Decoction.

[0081] Method: Random grouping, with 10 rats in each group, namely the blank group, the model group, the positive drug group, the medium-dose groups of Examples 1 - 2, the medium-dose groups of Comparative Examples 1 - 5, the high-dose, medium-dose, and low-dose groups of Example 3. In each modeling group, 1 ml of blood was taken from the tail of each rat on the 1st day, and blood was taken every other day for 5 consecutive times. On the 2nd day, the animals in the model group and the drug administration groups were intraperitoneally injected with cyclophosphamide at a dose of 40 mg / kg, and the injection was given every other day (the subsequent injection dose was 20 mg / kg) for 4 consecutive times;

[0082] Administration method: The model group started administration on the first day, and the blank and model groups were given the same volume of distilled water by gavage every day; the daily gavage doses were as follows: positive drug group: 1.242 g / kg, Examples 1-2 and Comparative Examples 1-5 groups: 2.970 g / kg, high-dose group of Example 3: 5.940 g / kg, medium-dose group of Example 3: 2.970 g / kg, low-dose group of Example 3: 1.485 g / kg, and all were administered for 21 days.

[0083] Administer the drug in the above manner. On the 21st day of administration, blood was taken from the orbital cavity for routine blood analysis and detection. The experimental results are shown in Table 3.

[0084] Table 3

[0085]

[0086] According to the experimental data of qi and blood deficiency in rats in Table 3 above, the combined fermented product prepared in the examples of the present invention can significantly improve each blood image index. The results of the immune function test show that it can significantly improve the blood image indexes of red blood cells RBC, white blood cells WBC, and hemoglobin HGB in rats with qi and blood deficiency caused by cyclophosphamide, significantly increase white blood cells, and all of the above indexes are significantly better than the model group; further verifying that the combined fermented product claimed in the present invention can enhance the erythroid hematopoietic function to achieve the effect of tonifying both qi and blood. Among the administration groups of combined fermented products with different ratios of substrates and different strains of bacteria, their functions are different, and the blood image indexes of Example 3 are the most significant.

[0087] Conclusion: The combined fermented product has the effects of tonifying qi and blood, improving bone marrow suppression, and increasing white blood cells, and it does not stagnate or hinder the spleen.

[0088] 4. Usage effects on patients with qi and blood deficiency syndrome

[0089] Inclusion criteria: 1) Can accept the determination of blood image indexes after medication. 2) Meet the following symptoms of qi and blood deficiency: dizziness, blurred vision, pale or sallow complexion, palpitations, insomnia, chest tightness, cold limbs, pale or thready and weak lips and nails, dry and falling hair, listlessness, fatigue and spontaneous sweating, loss of appetite, soreness and heaviness in the waist and legs, and decreased menstrual volume. 3) Chemotherapy and radiotherapy patients. 4) Surgical blood loss patients. 4) The elderly, the weak, and the population with low immunity. 5) Other patients with chronic diseases accompanied by qi and blood weakness.

[0090] Judgment basis: Marked effect: The blood image indexes are detected to be normal; the symptoms of qi and blood deficiency are significantly improved.

[0091] Effective: The blood image indexes are detected to tend to be normal, or at least one of the 3 indexes (RBC, WBC, HGB) is improved compared with before medication; the symptoms of qi and blood deficiency are improved.

[0092] Ineffective: The blood image indexes are detected as qi and blood deficiency indexes; the symptoms are not improved.

[0093] Administration regimen: Take the combined ferment prepared in Example 3 twice a day, on an empty stomach in the morning and evening, 30 - 50 ml each time. After taking the medicine continuously for 21 - 28 days, draw blood for testing and record the improvement of symptoms.

[0094] Results: The statistical results are shown in Table 4.

[0095] Table 4

[0096]

[0097] Among them, 5 ineffective patients are the elderly with long - term chronic underlying diseases. They took other therapeutic drugs during the medication period, making it difficult to exclude the interference of other drugs, but the fatigue state was improved. The chemotherapy and radiotherapy patients showed significant effects after taking the medicine, mostly effective, and a few were markedly effective. The patients with surgical blood loss showed obvious physical recovery after taking the medicine, and most of the blood picture indexes were markedly effective, and a few were effective.

[0098] Conclusion: The total effective rate for patients with qi and blood deficiency syndrome is 83.9%. It can improve qi and blood deficiency syndrome, increase blood picture indexes, and has the effect of increasing white blood cells.

[0099] In summary, the combined ferment provided by the present invention has the efficacy of supplementing both qi and blood, and is used for tonifying qi and blood deficiency, physical weakness, low immunity, weak spleen and stomach caused by chemotherapy, radiotherapy, surgery, old age and infirmity, postpartum, long - term illness, etc., and can rapidly increase white blood cells.

[0100] 5. Research on the effect of the combined ferment in nourishing blood and tranquilizing the mind to improve sleep

[0101] In this part of the research, the observation experiment of the number of spontaneous activities of mice was adopted, that is, to reflect the quiet sleep time, and the effects of Examples 1 - 3 and Comparative Examples 1 - 5 on the spontaneous activities of mice were investigated in combination with the growth situation, and the efficacy strengths among each group were compared.

[0102] Experimental animals: 120 clean - grade ICR male mice, with a body weight of 18 - 20 g.

[0103] Drugs: The combined ferments prepared in Examples 1 - 3 and Comparative Examples 1 - 5, positive drug: diazepam tablets.

[0104] Methods and administration: Randomly divide into groups, 10 mice in each group, namely the blank group, the positive drug group, the medium - dose groups of Examples 1 - 2, the medium - dose groups of Comparative Examples 1 - 5, the high - dose, medium - dose and low - dose groups of Example 3. Before administration and 1 h after the last administration, each group of mice was placed into a mouse spontaneous activity tester respectively. After adapting to the environment for 5 min, the instrument automatically recorded the number of spontaneous activities and standing times of the mice within 5 min;

[0105] Experimental environmental control conditions: The experimental site is ventilated and light-proof; room temperature is 23 ± 1 °C; humidity is 50 ± 5%; there is a 12 / 12 h light / dark cycle. Animals are adaptively fed for 5 - 7 days before the experiment. During the experiment, mice have free access to water and food.

[0106] The blank group was administered an equal volume of normal saline by gavage daily; the positive drug group: 1.25 mg / kg of diazepam suspension was given; for the groups of Examples 1 - 2 and Comparative Examples 1 - 5: 2.970 g / kg, the high-dose group of Example 3: 5.940 g / kg, the medium-dose group of Example 3: 2.970 g / kg, the low-dose group of Example 3: 1.485 g / kg. All were administered for 7 days, once a day, by oral gavage, and the dosage for each mouse was calculated based on 20 g. The experimental results are shown in Table 4.

[0107]

[0108] According to the experimental data in Table 5 above, the combined ferment prepared in the examples of the present invention can reduce the activity times of mice and increase the static sleep time. Compared with before administration, there are significant differences in each example group (P < 0.05), and among them, there are significant differences in the administration effects of the medium-dose groups of Examples 1 and 2, and the low-, medium-, and high-dose groups of Example 3 (P < 0.01). The mice administered with the drugs in each example group had smooth hair, obvious weight gain, good growth status, showed good mental and behavioral states. The results of the voluntary activity experiment showed that the combined ferment has a strong sedative effect on the sleep center, and thus shows a good effect in improving sleep.

[0109] Conclusion: The combined ferment can improve sleep quality by tonifying qi and nourishing blood and calming the mind.

[0110] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than a limitation on the protection scope of the present invention. Any simple modification or equivalent replacement of the technical solution of the present invention by those of ordinary skill in the art does not depart from the essence and scope of the technical solution of the present invention.

Claims

1. A combined ferment of ginseng, astragalus membranaceus and donkey-hide gelatin, characterized in that: The combined ferment is obtained by fermenting 1-3 parts of ginseng, 10-30 parts of astragalus membranaceus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1; the preparation method of the combined ferment comprises the following steps: (1) Weigh the ginseng and astragalus membranaceus in the formula amount, reflux and extract with water, combine the filtrates, concentrate, and reserve; (2) Crush the donkey-hide gelatin, add it to the filtrate in step (1) to dissolve, and then cool to obtain a mixed solution for reserve; (3) Add the bacterial powders of Lactobacillus bulgaricus and Lactobacillus plantarum to sterile physiological saline respectively to prepare bacterial suspensions, add the bacterial suspensions to the mixed solution in step (2), carry out fermentation culture, and sterilize to obtain the combined ferment; The temperature of the fermentation described in step (3) is 36-39 °C, and the time is 36-48 h.

2. A method for preparing the combined ferment described in claim 1, characterized in that: Comprising the following steps: (1) Weigh the ginseng and astragalus membranaceus in the formula amount, reflux and extract with water, combine the filtrates, concentrate, and reserve; (2) Crush the donkey-hide gelatin, add it to the filtrate in step (1) to dissolve, and then cool to obtain a mixed solution for reserve; (3) Add the bacterial powders of Lactobacillus bulgaricus and Lactobacillus plantarum to sterile physiological saline respectively to prepare bacterial suspensions, add the bacterial suspensions to the mixed solution in step (2), carry out fermentation culture, and sterilize to obtain the combined ferment; The temperature of the fermentation described in step (3) is 36-39 °C, and the time is 36-48 h.

3. The preparation method according to claim 2, wherein: The number of times of reflux extraction described in step (1) is 2-3 times, the time for each time is 1-2 h, and the water addition amount is 2-5 times the total mass of ginseng and astragalus membranaceus; the cooling temperature described in step (2) is 36-38 °C; the concentration of the bacterial suspension described in step (3) is 1-2 mg / mL.

4. Use of the combined ferment according to claim 1 in the preparation of a medicament for treating or preventing qi and blood deficiency syndrome.

5. The application according to claim 4, wherein: The qi and blood deficiency syndrome includes qi and blood deficiency, physical fatigue, low immunity or weak spleen and stomach caused by chemotherapy, radiotherapy, surgery, old age and infirmity, postpartum or long illness.

6. Use of the combined ferment according to claim 1 in the preparation of a medicament for treating or preventing leukopenia caused by myelosuppression.

7. Use of the combined ferment according to claim 1 in the preparation of a medicament or health product for improving sleep.

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