Medicinal and edible composition, medicinal and edible fermentation liquor, preparation method of medicinal and edible composition and application of medicinal and edible fermentation liquor in improvement of osteoporosis

Abstract

The invention provides a medicinal and edible composition, medicinal and edible fermentation liquor, a preparation method and application of the medicinal and edible composition and the medicinal and edible fermentation liquor in improvement of osteoporosis, and belongs to the technical field of microbial fermentation. The medicinal and edible composition comprises the following raw materials: radix astragali, rhizoma polygonati, raspberry, fructus lycii, polished round-grained rice and black bean coat. The medicinal and edible composition disclosed by the invention has the effects of invigorating spleen, replenishing qi and nourishing liver and kidney, and can promote osteoblast differentiation, reduce bone loss and enhance body energy metabolism, thereby effectively improving osteoporosis. In addition, the medicinal and edible composition disclosed by the invention is high in safety, capable of avoiding adverse reaction on a user, and beneficial to popularization. The invention also provides a medicinal and edible fermentation broth. The medicinal and edible fermentation broth comprises the following raw materials: a concentrated water extract of the medicinal and edible composition and the lactobacillus rhamnosus NKU FL1-8, the medicinal and edible homologous fermentation liquor can also effectively improve osteoporosis, and the effect of the medicinal and edible homologous fermentation liquor is superior to that of a concentrated water extract of a medicinal and edible homologous composition.

Description

Technical Field

[0001] The present invention belongs to the technical field of microbial fermentation, and in particular relates to a medicine-food homologous composition, a medicine-food homologous fermentation liquid, a preparation method thereof, and an application thereof in improving osteoporosis. Background Art

[0002] Osteoporosis is a pathological condition characterized by low bone mass and microstructural damage, leading to increased bone brittleness and susceptibility to fractures. The pathogenesis of osteoporosis is complex, and currently, clinical treatment primarily relies on medication to slow its progression. However, long-term drug dependence can lead to drug resistance and side effects such as gastrointestinal discomfort and liver and kidney damage. Therefore, developing a safe and effective alternative product is of great clinical value. Summary of the Invention

[0003] The purpose of the present invention is to provide a medicine-food composite, a medicine-food fermentation liquid, a preparation method thereof, and an application thereof in improving osteoporosis.

[0004] The invention provides a medicine and food composition, comprising the following raw materials in parts by weight: 5-30 parts of astragalus, 5-20 parts of polygonatum, 10-20 parts of raspberry, 10-50 parts of wolfberry, 5-20 parts of polished rice and 5-15 parts of black bean skin.

[0005] The present invention also provides a method for preparing the edible and medicinal composition described in the above scheme, comprising the following steps:

[0006] Astragalus root, polygonatum sibiricum, raspberry, wolfberry, polished rice, black bean skin and water are mixed, and the mixture is soaked, extracted and concentrated in sequence to obtain a medicine and food composition.

[0007] Preferably, the ratio of the total mass of the astragalus, polygonatum, raspberry, wolfberry, polished rice and black bean skin to the volume of water is 1g:3-5mL; the soaking time is 30min; the extraction temperature is 200-500°C; and the extraction time is 45min.

[0008] Preferably, the concentration of the medicine-food composition is 1 g / mL.

[0009] The present invention also provides a medicinal and edible fermentation liquid, the raw materials of which include a concentrated water extract of the medicinal and edible composition and Lactobacillus rhamnosus NKU FL1-8;

[0010] The medicine-food composition is the medicine-food composition described in the above scheme or the medicine-food composition prepared by the preparation method.

[0011] The present invention also provides a method for preparing the medicinal and edible fermentation liquid described in the above scheme, comprising the following steps:

[0012] Inoculating the concentrated water extract of the edible and medicinal composition with Lactobacillus rhamnosus NKU FL1-8 for fermentation to obtain a fermentation product;

[0013] The fermentation product is subjected to solid-liquid separation, and the liquid component is collected to obtain a medicine-food fermentation liquid.

[0014] Preferably, the effective viable count of the Lactobacillus rhamnosus NKU FL1-8 is 1×10 4 ~9×10 10 CFU / mL.

[0015] Preferably, the fermentation temperature is 35-38° C.; and the fermentation time is 24-48 hours.

[0016] The present invention also provides the use of the medicine-food composition described in the above scheme, the medicine-food composition prepared by the preparation method, the medicine-food fermentation liquid, or the medicine-food fermentation liquid prepared by the preparation method in preparing a drug for preventing and / or improving osteoporosis.

[0017] The present invention also provides the use of the edible and medicinal composition described in the above scheme, the edible and medicinal composition prepared by the preparation method, the edible and medicinal fermentation liquid, or the edible and medicinal fermentation liquid prepared by the preparation method in preparing products for improving bone density; the products include foods, health products, and medicines.

[0018] The present invention provides a medicinal and edible composition comprising the following raw materials in parts by weight: 5-30 parts of astragalus, 5-20 parts of polygonatum, 10-20 parts of raspberry, 10-50 parts of wolfberry, 5-20 parts of polished rice, and 5-15 parts of black bean skin. The medicinal and edible composition of the present invention has the effects of strengthening the spleen and replenishing qi, nourishing the liver and kidneys, and can promote osteoblast differentiation, reduce bone loss, and enhance the body's energy metabolism, thereby effectively improving osteoporosis. In addition, the medicinal and edible composition of the present invention is highly safe and can avoid adverse reactions in the recipient population, which is conducive to promotion. The present invention provides a safe and effective alternative product for improving osteoporosis, which has important clinical value.

[0019] The present invention also provides a medicine-food fermentation liquid, the raw materials of which include a concentrated water extract of the medicine-food composition and Lactobacillus rhamnosus NKU FL1-8; the medicine-food fermentation liquid can also effectively improve osteoporosis, and the effect is better than the concentrated water extract of the medicine-food composition described in the above scheme. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0021] Figure 1 This is a pie chart of the differential metabolites before and after fermentation;

[0022] Figure 2 The volcano plot of the differential metabolites before and after fermentation (the first 10), with significantly upregulated metabolites in red, significantly downregulated metabolites in blue, and non-significantly different metabolites in gray;

[0023] Figure 3 This is the match analysis graph of the differential metabolites (top 10) before and after fermentation;

[0024] Figure 4 The micro-CT analysis results of femur of rats in each group;

[0025] Figure 5 The results of HE staining and Trap staining of femoral tissues of rats in each group;

[0026] Figure 6 is the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels of rats in each group;

[0027] Figure 7 Comparison results of liver index of rats in each group, i.e. liver / body weight. DETAILED DESCRIPTION

[0028] The invention provides a medicine and food composition, comprising the following raw materials in parts by weight: 5-30 parts of astragalus, 5-20 parts of polygonatum, 10-20 parts of raspberry, 10-50 parts of wolfberry, 5-20 parts of polished rice and 5-15 parts of black bean skin.

[0029] The edible and medicinal composition of the present invention has the effects of strengthening the spleen and replenishing qi, nourishing the liver and kidneys, promoting osteoblast differentiation, reducing bone loss, and enhancing energy metabolism. Furthermore, the edible and medicinal composition of the present invention is highly safe and can avoid adverse reactions in the recipient population, facilitating its promotion.

[0030] In the present invention, Astragalus has the effects of strengthening the spleen and replenishing the middle, raising yang and lifting the sunken, benefiting the defense and consolidating the exterior, promoting diuresis and detoxifying and promoting tissue regeneration. It can promote osteoblast proliferation and differentiation and angiogenesis, show estrogen activity, prevent bone loss caused by estrogen deficiency, and maintain bone homeostasis.

[0031] In the present invention, Polygonatum sibiricum has the effects of nourishing yin and moistening the lungs, tonifying the spleen and replenishing qi, and nourishing the kidneys and replenishing essence. It can promote the differentiation of bone marrow mesenchymal stem cells into osteoblasts, promote bone formation, reduce the positive expression activity of osteocalcin and tartrate-resistant acid phosphatase, and promote fracture healing.

[0032] In the present invention, raspberry has the effects of solidifying essence and reducing urine, benefiting liver and kidney and improving eyesight, and promoting calcium salt deposition and collagen production.

[0033] In the present invention, wolfberry has the effects of nourishing the liver and kidney, improving eyesight and strengthening the kidney and bones, and promoting osteoblast differentiation.

[0034] In the present invention, the japonica rice has the effects of invigorating qi and strengthening spleen, eliminating thirst and stopping diarrhea, and can invigorate spleen and stomach, nourish the five internal organs, strengthen muscles and bones, promote blood circulation, and benefit essence and strengthen will.

[0035] In the present invention, the black bean skin has the effects of nourishing blood and dispelling wind, nourishing yin and blood, calming the liver and benefiting the kidneys, activating cartilage cells, promoting collagen production, inhibiting prostaglandin synthesis, and reducing joint swelling.

[0036] In the present invention, astragalus root is used to strengthen the spleen and replenish the middle, and is the monarch drug; polygonatum sibiricum and raspberry are used to nourish the kidney and liver, replenish essence and produce marrow, and are the minister drugs, enhancing the effect of astragalus root to strengthen the spleen and replenish qi, while also nourishing the liver and kidney; wolfberry and glutinous rice are used as adjuvant drugs to nourish yin, nourish the liver and strengthen the spleen, and have the effect of strengthening the spleen and replenishing qi, while also nourishing liver yin; black bean skin is sweet in taste and neutral in nature, and has the effect of harmonizing the various drugs, and is the guiding drug. The above combination conforms to the principle of "monarch, minister, assistant and guiding" compatibility and can play a corresponding intervention effect on the mechanism of osteoporosis.

[0037] The present invention also provides a method for preparing the edible and medicinal composition described in the above scheme, comprising the following steps:

[0038] Astragalus root, polygonatum sibiricum, raspberry, wolfberry, polished rice, black bean skin and water are mixed, and the mixture is soaked, extracted and concentrated in sequence to obtain a medicine and food composition.

[0039] As an embodiment, the ratio of the total mass of the astragalus, polygonatum, raspberry, wolfberry, polished rice and black bean skin to the volume of water is 1g:3-5mL; the soaking time is 30min; the extraction temperature is 200-500°C; and the extraction time is 45min.

[0040] As an embodiment, the concentration of the medicine-food composition is 1 g / mL.

[0041] The present invention also provides a medicine-food fermentation liquid, the raw materials of which include a concentrated water extract of the medicine-food composition and Lactobacillus rhamnosus NKU FL1-8; the medicine-food composition is the medicine-food composition described in the above scheme or the medicine-food composition prepared by the preparation method.

[0042] The edible and medicinal composition of the present invention is fermented by Lactobacillus rhamnosus NKU FL1-8, and organic acids and terpenes, including phytoestrogens such as biochanin and genistein, and small molecule compounds with antioxidant and anti-inflammatory properties, such as gingerol, brevifolia, hydroxylauric acid, and apigenin, are significantly upregulated. These substances have been previously documented to improve osteoporosis, osteoblast proliferation and differentiation, bone tissue regeneration, and injury repair. The edible and medicinal fermentation broth of the present invention can also effectively improve osteoporosis, with superior effects compared to the concentrated aqueous extract of the edible and medicinal composition.

[0043] The present invention also provides a method for preparing the medicinal and edible fermentation liquid described in the above scheme, comprising the following steps:

[0044] The concentrated water extract of the edible and medicinal composition is inoculated with Lactobacillus rhamnosus NKU FL1-8 to carry out fermentation to obtain a fermentation product; the fermentation product is subjected to solid-liquid separation, and the liquid component is collected to obtain a edible and medicinal fermentation liquid.

[0045] The invention first inoculates Lactobacillus rhamnosus NKUFL1-8 into the concentrated water extract of the medicine-food composition, performs fermentation, and obtains a fermentation product.

[0046] As an embodiment, the effective viable count of the Lactobacillus rhamnosus NKU FL1-8 is 1×10 4 ~9×10 10 CFU / mL; the rhamnosus casei bacteria NKU FL1-8 is deposited in the Guangdong Provincial Microbial Culture Collection Center with a deposit number of GDMCC No: 64053 and a deposit date of November 23, 2023; the rhamnosus casei bacteria NKU FL1-8 is inoculated in the form of a bacterial suspension; the inoculation amount of the rhamnosus casei bacteria NKU FL1-8 bacterial suspension accounts for 10% of the volume of the concentrated water extract of the medicine-food composition; the present invention has no special restrictions on the preparation method of the bacterial suspension, and conventional methods in the art can be used.

[0047] As an embodiment, the fermentation temperature is 35-38° C.; the fermentation time is 24-48 hours.

[0048] After obtaining the fermentation product, the present invention performs solid-liquid separation on the fermentation product, collects the liquid component, and obtains the medicine-food fermentation liquid.

[0049] As an embodiment, the solid-liquid separation includes centrifugation; the centrifugal speed is 5000 rpm; and the centrifugal time is 20 minutes.

[0050] After collecting the liquid components, the present invention further includes heating the liquid components in a water bath; the water bath heating temperature is 78 to 90° C.; the water bath heating time is 8 to 12 minutes; the water bath heating is used for sterilization and termination of the fermentation process.

[0051] The present invention also provides the use of the medicine-food composition described in the above scheme, the medicine-food composition prepared by the preparation method, the medicine-food fermentation liquid, or the medicine-food fermentation liquid prepared by the preparation method in preparing a drug for preventing and / or improving osteoporosis.

[0052] In the present invention, the prevention and / or improvement of osteoporosis includes promoting osteoblast differentiation, reducing bone loss, increasing trabecular area, improving femoral tissue structural stability, reducing osteoclast positive rate, increasing bone volume fraction, increasing trabecular number and reducing serum ALT and / or AST activity.

[0053] As an embodiment, the osteoporosis includes osteoporosis caused by estrogen deficiency; the estrogen deficiency is caused by ovarian removal.

[0054] As an embodiment, the dosage form of the drug includes lyophilized powder, liquid preparation or tablet.

[0055] The present invention also provides the use of the edible and medicinal composition described in the above scheme, the edible and medicinal composition prepared by the preparation method, the edible and medicinal fermentation liquid, or the edible and medicinal fermentation liquid prepared by the preparation method in preparing products for improving bone density; the products include foods, health products, and medicines.

[0056] To further illustrate the present invention, the following detailed description of a medicinal and edible composition, a medicinal and edible fermentation liquid, a preparation method, and an application in improving osteoporosis provided by the present invention is provided with reference to the accompanying drawings and examples. However, these should not be construed as limiting the scope of protection of the present invention.

[0057] Example 1 Preparation of concentrated aqueous extract of edible and medicinal composition

[0058] The raw materials of the medicine-food combination are: 5g of astragalus, 5g of polygonatum, 10g of raspberry, 10g of wolfberry, 5g of polished rice and 5g of black bean skin.

[0059] The preparation steps are as follows:

[0060] Astragalus, Polygonatum, raspberry, wolfberry, japonica rice and black bean skin are added to purified water with a mass-to-volume ratio of 3 times and soaked for 30 minutes. After decocting at 200-500°C for 45 minutes, the water extract is placed in a rotary evaporator and the volume is adjusted to 1g / mL to obtain a concentrated water extract of a medicine-food composition.

[0061] Example 2 Preparation of concentrated aqueous extract of edible and medicinal composition

[0062] The raw materials of the medicine-food combination are as follows: 30g of astragalus, 20g of polygonatum, 20g of raspberry, 50g of wolfberry, 20g of polished rice and 15g of black bean skin.

[0063] The preparation steps are as follows:

[0064] Astragalus, Polygonatum, raspberry, wolfberry, polished rice and black bean skin were added to purified water with a mass volume ratio of 5 times and soaked for 30 minutes. After decocting at 200-500°C for 45 minutes, the water extract was placed in a rotary evaporator and the volume was adjusted to 1g / mL to obtain a concentrated water extract of the medicine-food composition.

[0065] Example 3 Preparation of Food and Medicine Fermentation Broth

[0066] 1. Preparation of NKU FL1-8 bacterial suspension

[0067] Lactobacillus rhamnosus NKU FL1-8 was cultured in MRS liquid medium, the culture medium was centrifuged, and the precipitate was resuspended to obtain a bacterial suspension of Lactobacillus rhamnosus NKU FL1-8 with a concentration of 1×10 4 CFU / mL.

[0068] 2. The NKU FL1-8 bacterial suspension was inoculated into the extract of the edible and medicinal composition prepared in Example 1. The inoculation amount of the Lactobacillus rhamnosus NKU FL1-8 bacterial suspension accounted for 10% of the volume of the concentrated aqueous extract of the edible and medicinal composition. The fermentation was carried out at a temperature of 35 to 38° C. for 48 hours. The precipitate was then removed by centrifugation at 5000 rpm for 20 minutes. The supernatant was collected and heated in a water bath at 78 to 90° C. for 8 to 12 minutes.

[0069] Example 4 Preparation of Food and Medicine Fermentation Broth

[0070] 1. Preparation of NKU FL1-8 bacterial suspension

[0071] Lactobacillus rhamnosus NKU FL1-8 was cultured in MRS liquid medium, the culture medium was centrifuged, and the precipitate was resuspended to obtain a bacterial suspension of Lactobacillus rhamnosus NKU FL1-8 with a concentration of 9×10 10 CFU / mL.

[0072] 2. The NKU FL1-8 bacterial suspension was inoculated into the extract of the edible and medicinal composition prepared in Example 2. The inoculation amount of the Lactobacillus rhamnosus NKU FL1-8 bacterial suspension accounted for 10% of the volume of the concentrated aqueous extract of the edible and medicinal composition. The fermentation was carried out at a temperature of 35 to 38° C. for 24 hours. The precipitate was then removed by centrifugation at 5000 rpm for 20 minutes. The supernatant was collected and heated in a water bath at 78° C. to 90° C. for 8 to 12 minutes.

[0073] Test Example 1

[0074] Identification of the different components of the concentrated water extract of the medicinal and edible composition prepared in Example 1 and the medicinal and edible fermentation liquid prepared in Example 3

[0075] Non-target metabolomics analysis was carried out using LC / MS / MS technology to identify and compare the different components in the concentrated water extract of the medicine-food combination and the medicine-food fermentation broth. Figures 1 to 3 shown.

[0076] The results showed that the main types of differential metabolites before and after fermentation included organic acids and terpenes. Among them, phytoestrogens such as biochanin and genistein, and small molecule compounds with antioxidant and anti-inflammatory effects such as gingerol, pine pine, hydroxylauric acid, and apigenin were significantly upregulated after fermentation. The above substances have been proven in previous literature to have improvement effects on osteoporosis, osteoblast proliferation and differentiation, bone tissue regeneration, and damage repair.

[0077] Experimental Example 2: Improvement of the OVX Osteoporosis Rat Model by the Fermentation Broth of Lactobacillus rhamnosus NKU FL1-8-Medicine-Food Composition

[0078] Thirty rats were taken to establish an ovariectomy-induced osteoporosis model in rats, and a sham operation group (Sham), a model group (Model), a raloxifene positive control group (RLX), a concentrated water extract of the medicine-food composition group (TCM) and a medicine-food fermentation broth group (FTCM) were set up.

[0079] The rats were adaptively fed for one week before undergoing ovariectomy. After one week of recovery, the rats were given drugs by gavage once a day.

[0080] The sham operation group and the model group were administered with normal saline, respectively; the raloxifene positive control group was administered with raloxifene 6.3 mg / kg; the concentrated water extract of the medicine-food composition group was administered with 1.2 mL / kg of the concentrated water extract of the medicine-food composition; and the medicine-food fermentation liquid group was administered with 1.2 mL / kg of the medicine-food fermentation liquid.

[0081] After 12 weeks of intervention, the femoral tissues and cecal contents of the rats in each group were collected and stored at -80℃ for future use.

[0082] Micro-computed tomography (micro-CT) was used to examine the femoral tissue of rats, calculate bone density, observe the trabecular microstructure and generate 3D images, analyze the bone volume fraction (BV / TV) and trabecular number (Tb.N). The results of each group are shown in Figure 2. Figure 4 The femoral tissue was decalcified and embedded in paraffin, and then sliced ​​and stained with HE and Trap. Figure 5 ; The expression levels of corresponding biochemical indicators in rat serum were quantitatively detected using ALT and AST kits. The results are shown in Figure 6 .

[0083] Figure 4 The micro-CT results of each group showed that after intervention with the concentrated water extract of the edible and medicinal composition and the edible and medicinal fermentation liquid, the ovariectomy-induced osteoporosis in rats was improved. Compared with the concentrated water extract, the edible and medicinal fermentation liquid had a more significant improvement effect on indicators such as bone volume fraction and trabecular number.

[0084] Figure 5 HE staining results of femoral tissue of rats in each group showed that the trabecular structure in the femoral tissue of rats was significantly reduced, the bone density was reduced, the mature osteocytes were reduced, and the separation degree was high after ovariectomy. After intervention with the concentrated water extract and fermentation liquid of the medicine-food combination, the trabecular area increased and the femoral tissue structure was relatively stable. Trap staining results showed that the proportion of osteoclasts in the femoral tissue of rats increased after ovariectomy, while the osteoclast positive rate decreased after intervention, which was more significant in the medicine-food fermentation liquid (FTCM) group.

[0085] Figure 6 and Figure 7 The results of ALT, AST and liver index in each group showed that there was no statistical difference in ALT and AST activities between the groups after intervention with the concentrated water extract of the medicine-food combination and the medicine-food fermentation liquid compared with the blank group, indicating that no significant liver damage occurred after 12 weeks of oral administration of the medicine-food combination and the medicine-food fermentation liquid.

[0086] Although the above embodiment provides a detailed description of the present invention, it is only a part of the embodiments of the present invention, not all of the embodiments. People can also obtain other embodiments based on this embodiment without creativity, and these embodiments all fall within the scope of protection of the present invention.

Claims

1. A medicine-food composition, characterized in that: The preparation comprises the following raw materials in parts by weight: 5-30 parts of astragalus, 5-20 parts of polygonatum, 10-20 parts of raspberry, 10-50 parts of wolfberry, 5-20 parts of polished rice and 5-15 parts of black bean skin.

2. The method for preparing the edible and medicinal composition according to claim 1, characterized in that: The following steps are involved: Astragalus root, polygonatum sibiricum, raspberry, wolfberry, polished rice, black bean skin and water are mixed, and the mixture is soaked, extracted and concentrated in sequence to obtain a medicine and food composition.

3. The preparation method according to claim 2, characterized in that The ratio of the total mass of the astragalus, polygonatum, raspberry, wolfberry, polished rice and black bean skin to the volume of water is 1g:3-5mL; the soaking time is 30min; the extraction temperature is 200-500℃; and the extraction time is 45min.

4. The preparation method according to claim 2 or 3, characterized in that The concentration of the medicine-food composition is 1 g / mL.

5. A medicinal and edible fermentation liquid, characterized in that: The raw materials include concentrated water extract of the medicine-food combination and Lactobacillus rhamnosus NKU FL1-8; The medicine-food composition is the medicine-food composition according to claim 1 or the medicine-food composition prepared by the preparation method according to any one of claims 2 to 4.

6. The method for preparing the medicinal and edible fermentation liquid according to claim 5, characterized in that: The following steps are involved: Inoculating the concentrated water extract of the edible and medicinal composition with Lactobacillus rhamnosus NKU FL1-8 for fermentation to obtain a fermentation product; The fermentation product is subjected to solid-liquid separation, and the liquid component is collected to obtain a medicine-food fermentation liquid.

7. The preparation method according to claim 6, characterized in that: The effective viable count of the Lactobacillus rhamnosus NKU FL1-8 is 1×10 4 ~9×10 10 CFU / mL.

8. The preparation method according to claim 7, characterized in that The fermentation temperature is 35-38° C.; the fermentation time is 24-48 hours.

9. Use of the edible and medicinal composition of claim 1, the edible and medicinal composition prepared by the preparation method of any one of claims 2 to 4, the edible and medicinal fermentation liquid of claim 5, or the edible and medicinal fermentation liquid prepared by the preparation method of any one of claims 6 to 8 in the preparation of a drug for preventing and / or improving osteoporosis.

10. Use of the edible and medicinal composition of claim 1, the edible and medicinal composition prepared by the preparation method of any one of claims 2 to 4, the edible and medicinal fermentation liquid of claim 5, or the edible and medicinal fermentation liquid prepared by the preparation method of any one of claims 6 to 8 in preparing products for improving bone density; said products include foods, health products, and medicines.

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