Combined leavening of ginseng, astragalus membranaceus and donkey-hide gelatin as well as preparation method and application of combined leavening

Abstract

The invention provides a combined fermented product of ginseng, astragalus membranaceus and donkey-hide gelatin as well as a preparation method and application of the combined fermented product, and relates to the technical field of pharmaceutical preparations. The combined leavening is obtained by fermenting ginseng, astragalus membranaceus and donkey-hide gelatin, and the mass ratio of the ginseng to the astragalus membranaceus to the donkey-hide gelatin is (1-3): (10-30): 1. The composition of ginseng, astragalus membranaceus and colla corii asini is subjected to probiotic fermentation, macromolecular saponin is converted into aglycone or secondary saponin with smaller molecules, protein is split into peptide chains with smaller molecules, polysaccharide is split into sugar chains with smaller molecules, absorption and utilization of effective components of the medicine are facilitated, and the effect of tonifying qi and blood can be rapidly achieved.

Description

Technical Field

[0001] The present invention relates to the technical field of pharmaceutical preparations, and in particular to a combined fermentation product of ginseng, astragalus and donkey-hide gelatin, and a preparation method and application thereof. Background Art

[0002] Ginseng is the dried root and rhizome of Panax ginseng CAMeyer, a plant of the Araliaceae family. It tastes sweet, slightly bitter, and slightly warm. It has the functions of greatly replenishing vital energy, restoring pulse and consolidating deficiency, tonifying spleen and lung, promoting body fluid and nourishing blood, and calming the mind and improving intelligence. Ginseng greatly replenishes vital energy, focusing on restoring yang and rescuing deficiency, and replenishing the body's internal deficiency. The ginsenosides and polysaccharides contained in it are the main active ingredients. Astragalus is the dried root of Astragalus membranaceus (Fisch.) Bge. var. mongholicus (Bge .) Hsiao or Astragalus membranaceus (Fisch.) Bge., a plant of the Leguminosae family. It tastes sweet and warm. It has the functions of replenishing qi and raising yang, benefiting the body's defense and consolidating the exterior, promoting diuresis and reducing swelling, and supporting poison and promoting tissue regeneration. It mainly acts on the Ying and Wei, especially replenishing the Wei Qi, that is, the deficiency on the body's surface. It mainly contains astragaloside, astragaloside, etc. Donkey-hide gelatin is made by boiling and concentrating the skin of the donkey Equus asinus L., an equine animal. It tastes sweet and flat, and has the effects of nourishing blood and stopping bleeding, nourishing yin and moistening dryness, and calming the fetus. 80% of the ingredients in donkey-hide gelatin are collagen, amino acids and peptides, but it has the disadvantage of "nourishing and hindering the stomach". Ginseng is biased towards yin and replenishes the middle, while astragalus is biased towards yang and strengthens the exterior. The two medicines are combined, one yin and one yang, and they are used for each other to play the role of strengthening the body and replenishing qi. "Ben Cao Meng Quan" points out that ginseng regulates the middle, and astragalus strengthens the exterior, one exterior and one interior to achieve the function of replenishing qi both inside and outside. Donkey-hide gelatin can nourish blood and yin, moisten the lungs and stop bleeding. All three are strengthening medicines, taking into account qi, blood, yin, yang and internal organs, achieving the function of taking into account both the exterior and the interior, and replenishing qi and blood.

[0003] According to traditional Chinese medicine, Qi is the leader of blood, and blood is the mother of Qi. Clinically, Qi and blood deficiency syndrome manifests symptoms such as pale or sallow complexion, dizziness, vertigo, lack of breath, fatigue, chest tightness, insomnia, etc. In recent years, Qi and blood deficiency syndrome has become a common clinical disease. Chemotherapy, radiotherapy, surgery, old and weak, postpartum, long-term illness, etc. can all be seen in Qi and blood deficiency syndrome, with symptoms such as physical fatigue, low immunity, spleen and stomach weakness, bone marrow suppression, erythrocyte and leukocyte reduction, etc. Cardiovascular and cerebrovascular diseases, hypertension, coronary heart disease, heart failure, chronic obstructive pulmonary disease, diabetes, etc. are all accompanied by the occurrence of Qi deficiency syndrome. At present, the traditional Chinese medicine compound for the treatment of Qi and blood deficiency has many medicinal flavors, large dosage, slow onset, and most of the compound active ingredients are macromolecular substances with low bioavailability and low intestinal absorption, which will lead to a longer treatment cycle and the treatment obstacle of "deficiency cannot be supplemented".

[0004] The intestinal flora of patients with physical weakness is often disordered, and their digestion and absorption abilities are weakened. For example, the hydrolysis of donkey-hide gelatin collagen requires a large amount of protease, and patients with qi and blood deficiency have insufficient digestive enzymes and reduced digestive ability due to disordered intestinal flora. Therefore, taking donkey-hide gelatin not only increases the digestive burden, but also affects the absorption of the active ingredients of donkey-hide gelatin.

[0005] Probiotics are a type of microorganism that is beneficial to the host. They are widely present in the human intestinal tract and reproductive system and play an important role in the digestion and absorption of food and drugs. Using probiotics to transform saponins, polysaccharides, and peptides in traditional Chinese medicine can promote the conversion of effective ingredients into small molecules that are easier for the human body to absorb, and even produce new substances with stronger activity.

[0006] For example, Chinese patent CN112618691A discloses a Chinese medicine composition with the effect of replenishing qi and blood, and its preparation method and application, which are prepared as follows: S1: After washing and removing impurities from Chinese date seeds, lily, polygonatum, ginseng, carob, and jujube, crush them, add water and stir evenly to obtain a mixed solution; after adding a culture medium containing boulardii yeast, Lactobacillus plantarum, and Lactobacillus bulgaricus for mixed fermentation, after the fermentation is completed, filtering and decompression and concentration steps are performed to obtain a concentrated solution; S2: After washing and removing impurities from hawthorn, double red rose, wolfberry, malt, and dried ginger, extract them, and obtain a concentrated solution through a low-temperature decompression and concentration step; S3: Combine the above concentrated solutions, add auxiliary materials such as maltitol, and after boiling sugar and collecting paste steps, package them. The invention process can improve the extraction rate of effective ingredients in Chinese medicine, protect the effective ingredients in Chinese medicinal materials from being destroyed, degrade macromolecular substances to produce new polysaccharides, ginsenosides and other active ingredients, and at the same time reduce the toxic and side effects of Chinese medicine itself. Summary of the invention

[0007] Based on the prior art, the present invention provides a combined fermentation product of ginseng, astragalus and donkey-hide gelatin, and a preparation method and use thereof. By subjecting the combination of ginseng, astragalus and donkey-hide gelatin to probiotic fermentation, 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 drugs and can quickly exert the therapeutic effect of replenishing both qi and blood.

[0008] In order to achieve the above object, the present invention adopts the following technical solutions: On the one hand, the present invention provides a combined fermentation product of ginseng, astragalus and donkey-hide gelatin, wherein the combined fermentation product is obtained by fermenting ginseng, astragalus and donkey-hide gelatin, and the mass ratio of ginseng, astragalus and donkey-hide gelatin is 1-3:10-30:1.

[0009] The fermentation is carried out using probiotics; the probiotics are one or more of Lactobacillus and / or Bifidobacterium; The lactobacillus is selected from Lactobacillus bulgaricus and / or Lactobacillus plantarum.

[0010] Preferably, the probiotics are a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum, with a mass ratio of 1:1.

[0011] As some preferred embodiments, the combined fermentation product is obtained by fermenting 1-3 parts of ginseng, 10-30 parts of astragalus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0012] As a preferred embodiment, the combined fermentation product is obtained by fermenting 2 parts of ginseng, 25 parts of astragalus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0013] Ginseng mainly contains ginsenosides, polysaccharides and other active ingredients, which can greatly replenish the body's vitality and focus on replenishing the body's internal deficiency. Astragalus mainly replenishes the body's external deficiency, and mainly contains astragaloside, astragaloside, etc. Donkey-hide gelatin replenishes blood, and mainly contains collagen, amino acids and peptides.

[0014] The combined fermented product provided by the present invention includes donkey-hide gelatin, which is "greasy and stomach-injurious", and ginseng and astragalus contain ginsenosides, astragalus saponins, polysaccharides, etc. with large molecular weights, which are difficult to digest and absorb, and have low bioavailability. For the weak population with mostly disordered intestinal flora, "the deficiency cannot be supplemented", and a good tonic effect cannot be achieved. Therefore, it is necessary to use probiotics for in vitro fermentation and degradation to produce secondary low-molecular ginsenosides, astragalus saponins, small-molecule polysaccharides and other ingredients that are easier to absorb and more active.

[0015] The chemical components contained in the combined fermentation product are fermented and degraded by probiotics to produce rare ginsenosides, secondary astragaloside, secondary flavonoid glycosides, oligopeptides, amino acids, small molecular polysaccharides, etc. that are easier to absorb, more active, and have a smaller molecular weight. Probiotics ferment the ginseng, astragalus and donkey-hide gelatin composition in vitro, so that the chemical components contained in the Chinese medicine composition are fermented and degraded by probiotics in advance, reducing the digestive burden of patients with physical weakness. The use of probiotics for fermentation can significantly increase the content of rare ginsenosides Rh1, Rc, and Rg3 (S), and even produce some new active secondary metabolites.

[0016] The present invention also provides a method for preparing the combined fermentation product, comprising the following steps: (1) Weigh the amount of ginseng and astragalus used in the formula, add water and reflux to extract, combine the filtrate, concentrate and set aside; (2) Grinding donkey-hide gelatin, adding it to the filtrate from step (1) to dissolve, and then cooling it to obtain a mixed solution for use; (3) Add probiotic powders to sterile physiological saline to prepare bacterial suspensions, add the bacterial suspensions to the mixed solution of step (2), perform fermentation and sterilization, and obtain the combined fermentation product.

[0017] The reflux extraction in step (1) is performed 2-3 times, each time lasting 1-2 hours, and the amount of water added is 2-5 times the total mass of ginseng and astragalus, preferably 4 times.

[0018] The cooling temperature in the above step (2) is 36-38°C.

[0019] The fermentation temperature in step (3) is 36-39°C and the fermentation time is 36-48h.

[0020] The concentration of the bacterial suspension in step (3) above is 1-2 mg / mL, preferably 1 mg / mL.

[0021] The mass of the bacterial suspension in step (3) is 1%-2% of the total mass of ginseng, astragalus and donkey-hide gelatin; preferably 1%.

[0022] The present invention also provides the use of the combined fermentation product in preparing a product for treating or preventing qi and blood deficiency syndrome.

[0023] 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, postpartum or long-term illness.

[0024] The present invention also provides the use of the combined fermentation product in preparing a product for treating or preventing leukopenia.

[0025] The present invention also provides the use of the combined fermentation product in preparing a product for improving sleep.

[0026] The products include medicines, foods or health products.

[0027] Compared with the prior art, the present invention has the following beneficial effects: The combined fermentation provided by the present invention is based on the causes of intestinal flora disorder and weakened digestion and absorption capacity in patients with physical weakness, solves the treatment obstacle of "deficiency cannot be supplemented" in clinical Chinese medicine, and achieves the therapeutic purpose of rapid qi and blood supplementation. It has the effects of invigorating qi, replenishing blood, and activating 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 alleviate the symptoms of qi and blood deficiency, improve the body's immunity, and improve blood indicators such as red blood cells RBC, white blood cells WBC, hemoglobin HGB, platelet count PLT, etc. It is quickly absorbed and can be directly drunk without toxic side effects. DETAILED DESCRIPTION

[0028] The following will be combined with the tables in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0029] Hereinafter, examples and comparative examples of the present invention will be shown to explain the composition of the present invention in more detail, but the present invention is not limited thereto.

[0030] Example 1 A combined fermentation product of ginseng, astragalus and donkey-hide gelatin and its preparation method The combined fermented product is obtained by fermenting 1 part of ginseng, 10 parts of astragalus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0031] The specific preparation method is as follows: (1) Weigh the amount of ginseng and astragalus used in the formula, add twice the amount of water, reflux and extract twice, each time for 2 hours, combine the filtrates, concentrate and set aside; (2) Grind the donkey-hide gelatin in the prescribed amount, add it to the filtrate of step (1) to dissolve, and then cool it to 36° C. to obtain a mixed solution for use; (3) Lactobacillus bulgaricus powder and Lactobacillus plantarum powder are added to sterile physiological saline to prepare a 1 mg / mL bacterial suspension, which is added to the mixed solution of step (2) with a volume of the bacterial suspension being 1% of the volume of the mixed solution, and fermented at a temperature of 36° C. for 48 hours, followed by sterilization to obtain the fermented product of the composition.

[0032] Example 2 A combined fermentation product of ginseng, astragalus and donkey-hide gelatin and its preparation method The combined fermented product is obtained by fermenting 2 parts of ginseng, 30 parts of astragalus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0033] The specific preparation method is as follows: (1) Weigh the amount of ginseng and astragalus used in the formula, add 5 times the amount of water, reflux and extract 3 times, 1 hour each time, combine the filtrate, concentrate and set aside; (2) Grind the donkey-hide gelatin in the prescribed amount, add it to the filtrate of step (1) to dissolve, and then cool it to 38° C. to obtain a mixed solution for use; (3) Lactobacillus bulgaricus powder and Lactobacillus plantarum powder were added to sterile physiological saline to prepare a 2 mg / mL bacterial suspension, which was added to the mixed solution of step (2) with a volume of the bacterial suspension being 1% of the volume of the mixed solution. The mixture was fermented at a temperature of 39° C. for 36 hours and sterilized to obtain the fermented product of the composition.

[0034] Example 3 A combined fermentation product of ginseng, astragalus and donkey-hide gelatin and its preparation method The combined fermented product is obtained by fermenting 2 parts of ginseng, 25 parts of astragalus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:1.

[0035] The specific preparation method is as follows: (1) Weigh the amount of ginseng and astragalus used in the formula, add 4 times the amount of water, reflux and extract 3 times, each time for 1.5 hours, combine the filtrates, concentrate and set aside; (2) Grind the donkey-hide gelatin in the prescribed amount, add it to the filtrate of step (1) to dissolve, and then cool it to 37° C. to obtain a mixed solution for use; (3) Lactobacillus bulgaricus powder and Lactobacillus plantarum powder were added to sterile physiological saline to prepare a 1 mg / mL bacterial suspension, which was added to the mixed solution of step (2) with a volume of the bacterial suspension being 1% of the volume of the mixed solution. The mixture was fermented at a temperature of 38° C. for 40 hours and sterilized to obtain the fermented product of the composition.

[0036] Comparative Example 1 The difference from Example 3 is that only Lactobacillus bulgaricus is used. Other aspects are the same as Example 3.

[0037] Comparative Example 2 The difference from Example 3 is that only Lactobacillus plantarum is used. The other aspects are the same as those of Example 3.

[0038] Comparative Example 3 The difference from Example 3 is that no fermentation treatment is performed. Other aspects are the same as Example 3.

[0039] Comparative Example 4 The difference from Example 3 is that Astragalus is replaced by Codonopsis pilosula. The rest is the same as Example 3.

[0040] Comparative Example 5 The difference from Example 3 is that ginseng is replaced by codonopsis pilosula. The rest is the same as Example 3.

[0041] 1. Detection of rare ginsenosides Rh1, Rc, and Rg3(S) in combined fermentation products Methods: HPLC method was adopted, UV detector, reverse phase chromatographic column Agilent ZORBAX SB-C18 4.6Í250mm, 5μm; detection wavelength was 203nm, column temperature was 30℃, flow rate was 1ml / min, acetonitrile was used as mobile phase A, 0.05% volume of phosphoric acid water was used as mobile phase B, gradient elution was carried out, and the gradient elution conditions were 0-14min, 20-21%A; 14-24min, 21-22%A; 24-36min22-23%A; 36-52min, 23-25%A; 52-55min, 25-28%A; 55-78min, 28-30%A; 78-96min, 30-35%A; 96-115min, 35-60%A.

[0042] Reference substances: Ginsenosides Rh1, Rc, and Rg3(S) were purchased from Chengdu Munster Biotechnology Co., Ltd.

[0043] The test results are shown in Table 1 below.

[0044] Table 1 Rh1 (mg / 10ml) Rc (mg / 10ml) Rg3(S)(mg / 10ml) Example 1 3.998 8.511 0.087 Example 2 4.059 9.981 0.093 Example 3 4.540 11.751 0.106 Comparative Example 1 1.690 3.620 0.047 Comparative Example 2 2.210 6.579 0.064 Comparative Example 3 0.014 0.009 0.001 Comparative Example 4 2.580 4.465 0.055 Comparative Example 5 —— —— —— As shown in Table 1 above, under the action of different probiotic compositions, the contents of ginsenosides Rh1, Rc, and Rg3 (S) increased to varying degrees, among which 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 fermentation of a single strain, 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 indicates that fermentation treatment is beneficial to the dissolution of ginsenosides Rh1, Rc, and Rg3 (S).

[0045] 2. Detection of Cycloastragenol Content in Combined Fermentation Products Methods: HPLC-DAD-ELSD method was used, UV detector, reverse phase column Ultimate XB-C18 4.6Í250mm5μm; detection wavelength was 260nm, carrier gas flow rate: 3.2L / min, drift tube temperature: 110℃, gain: 1, flow rate: 1ml / min. Acetonitrile was used as mobile phase A, 0.1% volume formic acid water was used as mobile phase B, gradient elution was performed, the gradient elution conditions were 0-10min, 18%A; 10-32min, 18-35%A; 32-40min, 35%A; 40-57min, 35-53%A; 57-67min, 53-68%A.

[0046] Reference substance: Cycloastragenol, purchased from Chengdu Munster Biotechnology Co., Ltd.

[0047] The test results are shown in Table 2 below.

[0048] Table 2 Cycloastragenol (mg) Example 1 2.8536 Example 2 2.7569 Example 3 2.8715 Comparative Example 1 2.0341 Comparative Example 2 2.6593 Comparative Example 3 0.0305 Comparative Example 4 —— Comparative Example 5 2.068 As shown in Table 2 above, under the action of different probiotic compositions, the content of cycloastragenol, aglycone of astragaloside IV, a representative component of Astragalus, increased to varying degrees, among which Example 3 had the highest content of 2.8715 mg; in each comparative example, after fermentation with a single strain, the content of cycloastragenol increased compared with the unfermented group, but was significantly lower than that of the example group. This indicates that during the fermentation process, astragaloside IV may be enzymatically hydrolyzed by probiotics, thereby increasing the content of aglycone.

[0049] 3. Effect of combined fermentation products on leukocytosis in cyclophosphamide-induced myelosuppression rat model Experimental animals: 130 male SD rats, weighing 220±20 g.

[0050] Drug: combined fermentation products prepared in Examples 1-3 and Comparative Examples 1-5, positive drug: Baoyuan Decoction.

[0051] Methods: The animals were randomly divided into 10 groups, namely, blank group, model group, positive drug group, medium dose group of Examples 1-2, medium dose group of Comparative Examples 1-5, high dose group, medium dose group and low dose group of Example 3. Each modeling group was bled from the tail 1 ml / animal on the first day, and bled every other day for 5 consecutive times. The animals in the model group and drug-treated group were intraperitoneally injected with 40 mg / kg of cyclophosphamide on the second day, and injected every other day (the dose of subsequent injections was 20 mg / kg) for 4 consecutive times; Administration method: the modeling group was administered from the first day, and the blank and model groups were gavaged with an equal volume of distilled water daily; the daily gavage dosages were: positive drug group: 1.242 g / kg, Example 1-2 and Comparative Example 1-5 group: 2.970 g / kg, Example 3 high-dose group: 5.940 g / kg, Example 3 medium-dose group: 2.970 g / kg, Example 3 low-dose group: 1.485 g / kg, all were administered for 21 days.

[0052] The drugs were administered in the above manner, and blood was collected from the eye sockets on the 21st day of administration for routine blood analysis. The experimental results are shown in Table 3.

[0053] Table 3 Group Dosage g / kg <![CDATA[RBC*(10 12 / L)]]> <![CDATA[WBC*(10 9 / L)]]> HGB (g / L) Blank group - 8.312±0.513 5.750±1.680 141.210±4.700 Model Group - <![CDATA[8.050±0.290 # ]]> <![CDATA[2.040±0.450 ## ]]> <![CDATA[126.900±3.750 ## ]]> Positive control group 1.242 8.231±0.315 <![CDATA[3.250±0.690 * ]]> <![CDATA[136.210±3.120 * ]]> Example 1 Dose Group 2.970 <![CDATA[8.421±0.157 * ]]> <![CDATA[4.613±0.367 ** ]]> <![CDATA[136.349±2.163 * ]]> Example 2 Dose Group 2.970 <![CDATA[8.534±0.247 ** ]]> <![CDATA[4.751±0.329 ** ]]> <![CDATA[138.156±2.893 * ]]> Example 3 Low dose group 1.485 <![CDATA[8.676±0.364 ** ]]> <![CDATA[4.170±0.680 ** ]]> <![CDATA[139.110±3.690 * ]]> Example 3: Dose group 2.970 <![CDATA[8.755±0.299 ** ]]> <![CDATA[5.041±0.45 ** ]]> <![CDATA[142.180±3.411 ** ]]> Example 3 High dose group 5.940 <![CDATA[8.930±0.417 ** ]]> <![CDATA[5.787±0.580 ** ]]> <![CDATA[146.210±5.210 ** ]]> Comparative Example 1 Middle Dose Group 2.970 8.247±0.349 <![CDATA[3.342±0.219 * ]]> <![CDATA[132.897±2.547 * ]]> Comparative Example 2 Middle Dose Group 2.970 <![CDATA[8.305±0.127 * ]]> <![CDATA[3.897±0.310 * ]]> <![CDATA[135.457±4.016 * ]]> Comparative Example 3 Middle Dose Group 2.970 8.150±0.310 2.340±0.650 129.490±3.790 Comparative Example 4 Middle Dose Group 2.970 8.171±0.241 2.716±0.325 128.384±2.514 Comparative Example 5 Middle Dose Group 2.970 <![CDATA[8.332±0.352 * ]]> 2.698±0.360 131.109±3.056 Note: Compared with the blank group, # P<0.05, ## P<0.01; compared with the model group, * P<0.05, ** P<0.01.

[0054] According to the experimental data of Qi and blood deficiency in rats in Table 3 above, the combined fermentation prepared by the embodiment of the present invention can significantly improve various blood image indicators. The results of the immune function test show that the blood image indicators of erythrocytes RBC, white blood cells WBC, and hemoglobin HGB in rats with Qi and blood deficiency caused by cyclophosphamide can be significantly improved, and white blood cells are significantly increased. The above indicators are significantly better than the model group; further verifying that the combined fermentation claimed by the present invention can enhance the red blood cell hematopoietic function to achieve the function of Qi and blood supplementation. In the administration groups of different proportions of substrates and different strains of combined fermentation products, their functions are different, and the blood image indicators of Example 3 are the most significant.

[0055] Conclusion: The combined fermented product has the effects of replenishing qi and blood, improving bone marrow suppression, and increasing white blood cells, and it does not cause stagnation and hinder the spleen.

[0056] Inclusion criteria: 1) Acceptable blood test after medication. 2) Meet the following symptoms of Qi and blood deficiency: dizziness, blurred vision, pale or sallow complexion, palpitations and insomnia, chest tightness, cold limbs, pale lips and nails or weak pulse, dry hair, hair loss, shortness of breath, lazy speech, fatigue and spontaneous sweating, loss of appetite, back pain and heavy legs, and decreased menstrual volume. 3) Patients undergoing chemotherapy and radiotherapy. 4) Patients with surgical bleeding. 4) Elderly and weak people with low immunity. 5) People with other chronic diseases and weak Qi and blood.

[0057] Judgment basis: Markedly effective: blood index tests are normal; symptoms of qi and blood deficiency are significantly improved.

[0058] Effective: Blood index tests tend to be normal, or at least one of the three indicators (RBC, WBC, HGB) is improved compared with before medication; symptoms of qi and blood deficiency are improved.

[0059] Ineffective: Blood test showed Qi and blood deficiency; symptoms did not improve.

[0060] Dosage regimen: Take the combined fermentation product prepared in Example 3 twice a day, morning and evening on an empty stomach, 30-50 ml / time. After taking the medicine for 21-28 days, draw blood for testing and record the improvement of symptoms.

[0061] Results: The statistical results are shown in Table 4.

[0062] Table 4

[0063] Among them, 5 ineffective patients were elderly people with long-term chronic underlying diseases. They took other therapeutic drugs at the same time during the medication, and it was difficult to rule out the interference of other drugs, but the fatigue state was improved. The effect of the drug on patients undergoing chemotherapy and radiotherapy was significant, most of them were effective, and some were markedly effective. After taking the drug, patients with surgical bleeding recovered significantly, and most of the blood indicators were markedly effective, and some were effective.

[0064] Conclusion: The total effective rate for patients with qi and blood deficiency is 83.9%. It can improve qi and blood deficiency, increase blood indexes, and has the effect of increasing white blood cells.

[0065] In summary, the combined fermentation product provided by the present invention has the effect of replenishing both qi and blood, and is used to replenish qi and blood deficiency, physical fatigue, low immunity, weak spleen and stomach, etc. in chemotherapy, radiotherapy, surgery, the elderly and the weak, postpartum, long-term illness, and has the effect of quickly increasing white blood cells.

[0066] 5. Study on the effect of combined fermentation products on nourishing blood, calming nerves and improving sleep This part of the study adopts an observation experiment on the number of spontaneous activities of mice, which reflects the static sleep time, and examines the effects of Examples 1-3 and Comparative Examples 1-5 on the spontaneous activities of mice in combination with their growth conditions, and compares the strength of the drug efficacy between the groups.

[0067] Experimental animals: 120 clean-grade ICR mice, male, weighing 18-20 g.

[0068] Drug: combined fermentation products prepared in Examples 1-3 and Comparative Examples 1-5; positive drug: diazepam tablets.

[0069] Method and administration: The mice were randomly divided into 10 groups, namely, blank group, positive drug group, medium dose group of Example 1-2, medium dose group of Comparative Example 1-5, high dose group, medium dose group and low dose group of Example 3. Before administration and 1 hour after the last administration, each group of mice was placed in a mouse spontaneous activity tester. After acclimating to the environment for 5 minutes, the instrument automatically recorded the number of spontaneous activities and standing times of the mice within 5 minutes; Experimental environment control conditions: The experimental site was ventilated and dark; the room temperature was 23±1℃; the humidity was 50±5%; the light and dark cycle was 12 / 12h. The animals were adaptively raised for 5-7 days before the experiment. During the experiment, the mice were free to drink water and eat.

[0070] The blank group was gavaged with an equal volume of saline every day; the positive drug group: 1.25 mg / kg diazepam suspension was given, the Example 1-2 and Comparative Example 1-5 groups: 2.970 g / kg, the Example 3 high-dose group: 5.940 g / kg, the Example 3 medium-dose group: 2.970 g / kg, the Example 3 low-dose group: 1.485 g / kg, all were given for 7 days, once a day, oral gavage, and each mouse was given 20 g. The experimental results are shown in Table 4.

[0071] Table 5 Effects of combined fermentation products on the number of mouse activities ( ±S) Group Dose / kg Number of activities before medication (n) Number of activities after 7 days of administration (n) Blank group - 156.0±12.61 159.1±12.69 Positive control group 1.250mg 175.4±9.89 <![CDATA[142.4±16.41 ** ]]> Example 1 Dose Group 2.970g 165.4±14.60 <![CDATA[137.0±12.26 ** ]]> Example 2 Dose Group 2.970g 155.3±13.55 <![CDATA[130.7±13.52 ** ]]> Example 3 Low dose group 1.485g 157.9±12.60 <![CDATA[132.0±12.35 ** <!-- 7 -->]]> Example 3: Dose group 2.970g 168.7±12.35 <![CDATA[128.8±11.93 ** ]]> Example 3 High dose group 5.940g 158.5±11.95 <![CDATA[123.0±11.70 ** ]]> Comparative Example 1 Middle Dose Group 2.970g 154.8±11.75 148.5±11.84 Comparative Example 2 Middle Dose Group 2.970g 178.2±12.04 <![CDATA[156.0±12.05 * ]]> Comparative Example 3 Middle Dose Group 2.970g 166.3±12.11 156.8±12.53 Comparative Example 4 Middle Dose Group 2.970g 158.7±11.63 141.7±11.88 Comparative Example 5 Middle Dose Group 2.970g 161.5±12.04 145.1±11.72 Note: Compared with before administration, * P<0.05, ** P<0.01 According to the experimental data in Table 5 above, the combined fermentation prepared in the embodiment of the present invention can reduce the number of activities of mice and increase the static sleep time. Compared with before administration, each embodiment group has significant differences (P<0.05), among which the medium dose groups in embodiments 1 and 2, and the low, medium and high dose groups in embodiment 3 have significant differences in the administration effect (P<0.01). The mice given the drugs in each embodiment group have smooth fur, obvious weight gain, good growth status, and good mental and behavioral status. The results of the autonomous activity experiment show that the combined fermentation has a strong sedative effect on the sleep center, and therefore shows a good effect on improving sleep.

[0072] Conclusion: The combined fermented products can improve sleep quality by replenishing qi and blood and soothing the mind.

[0073] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A combined fermentation product of ginseng, astragalus and donkey-hide gelatin, characterized in that: The combined fermented product is obtained by fermenting ginseng, astragalus and donkey-hide gelatin, and the mass ratio of ginseng, astragalus and donkey-hide gelatin is 1-3:10-30:1; The fermentation is carried out using probiotics; the probiotics are one or more of lactobacillus and / or bifidobacterium.

2. The combined fermentation product according to claim 1, characterized in that: The lactobacillus is selected from Lactobacillus bulgaricus and / or Lactobacillus plantarum.

3. The combined fermentation product according to claim 2, characterized in that: The probiotics are a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum, with a mass ratio of 1:

1.

4. The combined fermentation product according to claim 1, characterized in that: The combined fermented product is obtained by fermenting 1-3 parts of ginseng, 10-30 parts of astragalus and 1 part of donkey-hide gelatin with a mixture of Lactobacillus bulgaricus and Lactobacillus plantarum in a mass ratio of 1:

1.

5. A method for preparing the combined fermentation product according to any one of claims 1 to 4, characterized in that: The steps include: (1) Weigh the amount of ginseng and astragalus used in the formula, add water and reflux to extract, combine the filtrate, concentrate and set aside; (2) Grinding donkey-hide gelatin, adding it to the filtrate from step (1) to dissolve, and then cooling it to obtain a mixed solution for use; (3) Add probiotic powders to sterile physiological saline to prepare bacterial suspensions, add the bacterial suspensions to the mixed solution of step (2), perform fermentation and sterilization, and obtain the combined fermentation product.

6. The preparation method according to claim 5, characterized in that: The number of reflux extractions in step (1) is 2-3 times, each time lasting 1-2 hours, and the amount of water added is 2-5 times the total mass of ginseng and astragalus; the cooling temperature in step (2) is 36-38° C.; the fermentation temperature in step (3) is 36-39° C., and the fermentation time is 36-48 hours; the concentration of the bacterial suspension in step (3) is 1-2 mg / mL.

7. Use of the combined fermentation product according to any one of claims 1 to 4 in the preparation of medicines, health products or foods for treating or preventing qi and blood deficiency syndrome.

8. The use according to claim 7, characterized in that: 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, postpartum or long-term illness.

9. Use of the combined fermentation product according to any one of claims 1 to 4 in the preparation of medicines, health products or foods for treating or preventing leukopenia caused by bone marrow suppression.

10. Use of the combined fermentation product according to any one of claims 1 to 4 in the preparation of sleep-improving medicines, health products or foods.

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