A mulberry morinda officinalis cistanche and herba comfrey compound preparation for tonifying kidney, and a preparation method and application thereof

Abstract

The present application relates to the technical field of traditional Chinese medicine preparation, and particularly relates to mulberry and cistanche-cynomorium compound preparation for tonifying kidney, and a preparation method and application thereof. The present application provides mulberry and cistanche-cynomorium compound preparation for tonifying kidney, which comprises the following components: mulberry extract 30 wt%, desert cistanche 25 wt%, cynomorium 20 wt%, kidney 20 wt% and astragalus 5 wt%; the kidney is sheep kidney or dog kidney. The mulberry and cistanche-cynomorium compound preparation for tonifying kidney provided by the present application is prepared by compounding and combining five components of mulberry extract, desert cistanche, cynomorium, sheep kidney and astragalus in specific contents, further combined with lactobacillus rhamnosus and bifidobacterium longum, and fermented with the combination of mulberry extract, desert cistanche, cynomorium and astragalus, so that the good effect of warming yang and tonifying kidney is achieved, and the gastrointestinal burden is not increased, and the preparation is suitable for people with weak spleen and stomach to take, and the warming yang and tonifying kidney effect of people with weak spleen and stomach is ensured.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine preparation technology, and in particular to a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney, its preparation method and application. Background Technology

[0002] With the continuous exploration of the field of warming and tonifying the kidneys in modern traditional Chinese medicine, a large number of kidney-tonifying Chinese herbs have been developed. These herbs can provide precise physiological function improvement and anti-aging support for different constitutions. For example, a combination of herbs such as deer antler, epimedium, eucommia, and psoralea can effectively improve symptoms such as aversion to cold and cold limbs, frequent urination at night, soreness of the waist and knees, as well as impotence and premature ejaculation; a combination of herbs such as rehmannia, wolfberry, privet fruit, eclipta prostrata, and donkey-hide gelatin can effectively relieve symptoms such as dry mouth and throat, hot flashes and night sweats, dizziness and tinnitus, and premature graying of hair; a combination of herbs such as dodder seed, schisandra fruit, euryale seed, and cornus fruit can not only tonify yang and nourish yin, but also consolidate essence, reduce urination, and stop diarrhea, making it suitable for those with deficiency of both yin and yang.

[0003] While the aforementioned kidney-tonifying Chinese medicines are indeed effective, there are some significant problems in their application. For example, the yin-nourishing and blood-tonifying properties of herbs like Rehmannia glutinosa and donkey-hide gelatin can easily lead to bloating and indigestion in individuals with weak spleen and stomach. They usually need to be used in combination with qi-regulating and spleen-strengthening herbs such as tangerine peel and amomum villosum to aid absorption. Therefore, developing a new kidney-tonifying Chinese medicine is of great significance to the development of the field of warming yang and tonifying the kidneys. Summary of the Invention

[0004] In view of this, the present invention provides a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney, its preparation method and application. The compound preparation of mulberry, cistanche and cynomorium provided by the present invention has good effects of warming yang and tonifying the kidney, and is suitable for people with weak spleen and stomach.

[0005] This invention provides a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney, comprising the following components: 30wt% mulberry extract, 25wt% desert cistanche, 20wt% cynomorium, 20wt% kidney, and 5wt% astragalus; wherein the kidney is sheep kidney or dog kidney; the preparation method of the mulberry extract includes the following steps: mixing mulberry, ethanol, water, and acid under acidic conditions for extraction, followed by solid-liquid separation to obtain the mulberry extract; the preparation method of the compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney includes the following steps: (1) Mulberry extract, desert cistanche, cynomorium, astragalus and sterile water are mixed and sterilized to obtain a fermentation substrate. Then, the fermentation substrate and lactic acid bacteria are mixed and fermented and then freeze-dried to obtain a fermentation powder. The lactic acid bacteria are Lactobacillus rhamnosus and Bifidobacterium longum. The mass ratio of Lactobacillus rhamnosus and Bifidobacterium longum is 10:8~12. (2) The kidneys are dried by baking with firewood and tiles and then ground into powder to obtain kidney powder; (3) Mix the fermented powder and kidney powder to obtain the mulberry, cistanche and cynomorium compound preparation for tonifying the kidney; there is no requirement for the time order of steps (1) and (2).

[0006] Preferably, the mass ratio of mulberry to ethanol is 100:23~28; and the mass ratio of mulberry to water is 100:200~240.

[0007] Preferably, the pH value of the acidic conditions is 5.5 to 6.

[0008] Preferably, the extraction temperature is 45-56 degrees Celsius, and the holding time is 1.5-1.8 hours; the extraction is carried out in a water bath.

[0009] Preferably, the Astragalus membranaceus is Astragalus sinensis var. sinensis.

[0010] This invention also provides a method for preparing the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney as described above, comprising the following steps: (1) Mulberry extract, desert cistanche, cynomorium, astragalus and sterile water are mixed and sterilized to obtain a fermentation substrate. Then, the fermentation substrate and lactic acid bacteria are mixed and fermented and then freeze-dried to obtain a fermentation powder. The lactic acid bacteria are Lactobacillus rhamnosus and Bifidobacterium longum. The mass ratio of Lactobacillus rhamnosus and Bifidobacterium longum is 10:8~12. (2) The kidneys are dried by baking with firewood and tiles and then ground into powder to obtain kidney powder; (3) Mix the fermented powder and kidney powder to obtain the mulberry, cistanche and cynomorium compound preparation for tonifying the kidney; there is no requirement for the time order of steps (1) and (2).

[0011] The present invention also provides the application of the mulberry, cistanche, and cynomorium compound preparation described above for tonifying the kidney in the preparation of kidney-tonifying drugs.

[0012] Compared with the prior art, the present invention has achieved the following beneficial effects: This invention provides a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys. Through the combination of five components—mulberry extract, desert cistanche, cynomorium, sheep kidney, and astragalus—and further combined with *Lactobacillus rhamnosus* and *Bifidobacterium longum*, the mulberry extract, desert cistanche, cynomorium, and astragalus are fermented together, achieving a good warming and tonifying effect on the kidneys. This invention further enhances the synergistic effect by controlling the dosage of the above components, thus further improving the warming and tonifying effect on the kidneys. Furthermore, *Lactobacillus rhamnosus* and *Bifidobacterium longum*, as intestinal probiotics, ensure that this compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys does not burden the gastrointestinal tract, making it suitable for those with weak spleen and stomach, thus guaranteeing the warming and tonifying effect on the kidneys for those with weak spleen and stomach. Attached Figure Description

[0013] To more clearly illustrate the technical solutions of this invention, the accompanying drawings used in the embodiments of this invention or in the prior art are briefly described below. For those skilled in the art, other drawings can be derived from the following drawings without creative effort, and all such drawings are within the protection scope of this invention.

[0014] Figure 1 A comparative graph showing the testosterone levels in rats that took the mulberry, cistanche, and cynomorium compound preparations used for tonifying the kidneys, as described in Examples 1-5 and Comparative Examples 1-10. Detailed Implementation

[0015] This invention provides a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney, comprising the following components: 30wt% mulberry extract, 25wt% desert cistanche, 20wt% cynomorium, 20wt% kidney, and 5wt% astragalus; wherein the kidney is sheep kidney or dog kidney.

[0016] The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney provided by this invention includes 30 wt% mulberry extract; the preparation method of the mulberry extract may include the following steps: mixing mulberry, ethanol, water and acid, extracting under acidic conditions, and then separating the solid and liquid to obtain the mulberry extract.

[0017] In this invention, the mass ratio of mulberry to ethanol can be 100:23 to 28, specifically 100:24, 100:25, 100:26 or 100:27.

[0018] In this invention, the mass ratio of mulberry to water can be 100:200~240, specifically 100:210, 100:220 or 100:230.

[0019] In this invention, the water can be deionized water.

[0020] In this invention, the pH value of the acidic conditions can be 5.5 to 6, specifically 5.6, 5.7, 5.8 or 5.9.

[0021] In this invention, the acid can be acetic acid. This invention achieves the aforementioned acidic conditions by adding an acid.

[0022] In this invention, the extraction temperature can be 45~56 degrees Celsius, specifically 47 degrees Celsius, 49 degrees Celsius, 51 degrees Celsius, 53 degrees Celsius or 55 degrees Celsius, and the heat preservation time can be 1.5~1.8 hours, specifically 1.6 hours or 1.7 hours.

[0023] In this invention, the extraction can be performed under water bath conditions.

[0024] In this invention, the solid-liquid separation can be filtration.

[0025] The kidney-tonifying compound preparation of mulberry, cistanche, and cynomorium provided by this invention includes 25 wt% desert cistanche; the desert cistanche can be desert cistanche from Alashan, Inner Mongolia.

[0026] The kidney-tonifying compound preparation of mulberry, cistanche, and cynomorium provided by this invention includes 20 wt% cynomorium; the cynomorium can be cynomorium from Alashan, Inner Mongolia.

[0027] The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney provided by this invention includes 20 wt% kidney; the kidney can be used in the form of kidney powder.

[0028] The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney provided by this invention includes 5 wt% astragalus; the astragalus can be Astragalus membranaceus from Hunyuan, Shanxi.

[0029] This invention also provides a method for preparing the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney as described above, comprising the following steps: (1) Mulberry extract, desert cistanche, cynomorium, astragalus and sterile water are mixed and sterilized to obtain a fermentation substrate. Then the fermentation substrate is mixed with lactic acid bacteria and fermented and then freeze-dried to obtain fermentation powder. (2) The kidneys are dried by baking with firewood and tiles and then ground into powder to obtain kidney powder; (3) Mix the fermented powder and kidney powder to obtain the mulberry, cistanche and cynomorium compound preparation for tonifying the kidney; there is no requirement for the time order of steps (1) and (2).

[0030] This invention involves sterilizing a mixture of mulberry extract, desert cistanche, cynomorium, astragalus, and sterile water to obtain a fermentation substrate. This substrate is then mixed with lactic acid bacteria for fermentation and freeze-dried to obtain a fermented powder. In this invention, the desert cistanche, cynomorium, and astragalus can be pretreated independently before use; the pretreatment may include the following steps: grinding the desert cistanche, cynomorium, or astragalus into powder.

[0031] In this invention, the mass ratio of Cynomorium songaricum to sterile water can be 1:20~26, specifically 1:21, 1:22, 1:23, 1:24 or 1:25.

[0032] In this invention, the sterilization can be high-temperature sterilization, and the temperature of the high-temperature sterilization can be 120~122 degrees Celsius, specifically 121 degrees Celsius; the sterilization holding time can be 10~20 minutes, specifically 15 minutes.

[0033] In this invention, the mass ratio of the fermentation substrate to the lactic acid bacteria can be 100:5~8, specifically 100:6 or 100:7.

[0034] In this invention, the lactic acid bacteria can be *Lactobacillus rhamnosus* and *Bifidobacterium longum*; the preservation number of *Lactobacillus rhamnosus* is ATCC7596; the preservation number of *Bifidobacterium longum* is CICC 6188; the mass ratio of *Lactobacillus rhamnosus* to *Bifidobacterium longum* can be 10:8~12, specifically 10:9, 10:10, or 10:11. Both *Lactobacillus rhamnosus* and *Bifidobacterium longum* used in this invention are commercially available products, obtained through commercial purchase. Specifically, they were purchased from the American Center for Type Culture Collection, and *Bifidobacterium longum* was purchased from the China Industrial Microbiological Culture Collection Center.

[0035] In this invention, the lactic acid bacteria can be pre-cultured before use; the pre-culture may include the following steps: inoculating the lactic acid bacteria into MRS medium and activating them in an anaerobic environment at 37 degrees Celsius for 18 hours, and then transferring them to a 10L fermenter for expansion culture at 37 degrees Celsius for 24 hours.

[0036] In this invention, the lactic acid bacteria can be used in the form of a lactic acid bacteria liquid; the viable count of Bifidobacterium longum in the lactic acid bacteria liquid can be 1×10⁻⁶. 9 ~1.5×10 9 The viable count of Lactobacillus rhamnosus can be 1×10⁻⁶ CFU / mL. 9 ~1.5×10 9 CFU / mL.

[0037] In this invention, the fermentation temperature can be 37 degrees Celsius, and the heat preservation time can be 60 to 90 hours, specifically 66 hours, 72 hours, 78 hours or 84 hours.

[0038] In this invention, the freeze-drying process may further include pre-freezing; the pre-freezing temperature may be -40 to -30 degrees Celsius, and the holding time may be 1 to 3 hours.

[0039] In this invention, the freeze-drying temperature can be -30 to -20 degrees Celsius, and the holding time can be 12 to 18 hours.

[0040] This invention involves drying the kidneys using firewood and roof tiles, then grinding them into powder to obtain kidney powder. The method of drying using the roof tiles is not specifically required; conventional methods in the art are sufficient.

[0041] After obtaining the fermented powder and kidney powder, the present invention mixes the fermented powder and kidney powder to obtain the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney. In the present invention, the mixing can be by grinding.

[0042] The present invention also provides the application of the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney as described in the above-described scheme, or the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney obtained by the preparation method described in the above-described scheme, in the preparation of kidney-tonifying drugs.

[0043] In this invention, the dosage form of the kidney-tonifying drug may include pills, elixirs, tablets, or capsules.

[0044] To further illustrate the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings and embodiments.

[0045] In a specific embodiment of the present invention, the pre-culture steps of lactic acid bacteria are as follows: Lactobacillus rhamnosus ATCC7596 and Bifidobacterium longum CICC 6188 are inoculated into MRS medium and activated under anaerobic conditions at 37 degrees Celsius for 18 hours. Then, they are transferred to a 10L fermenter for expansion culture at 37 degrees Celsius for 24 hours to obtain a lactic acid bacteria culture, wherein the viable count of Bifidobacterium longum is 1.3 × 10⁻⁶. 9 The viable count of *Lactobacillus rhamnosus* was 1.2 × 10⁻⁶ CFU / mL. 9 CFU / mL.

[0046] Example 1: This embodiment prepares a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney, with the following components: 30g mulberry extract, 25g desert cistanche, 20g cynomorium, 20g dog kidney, and 5g astragalus. The mulberry extract is prepared as follows: Mulberry, ethanol, deionized water, and acetic acid are mixed at a mass ratio of 100:25 and 100:210, and extracted in a 50°C water bath for 1.5 hours under acidic conditions (pH 5.8). The mixture is then filtered to obtain the mulberry extract. The desert cistanche is *Cistanche deserticola* from Alashan, Inner Mongolia; the cynomorium is *Cistanche cynomorium* from Alashan, Inner Mongolia; and the astragalus is *Astragalus membranaceus* from Hunyuan, Shanxi.

[0047] This embodiment describes the preparation of a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys. The specific steps are as follows: (1) Before use, Cistanche deserticola, Cynomorium songaricum, and Astragalus membranaceus were ground into powder. Then, mulberry extract, Cistanche deserticola, Cynomorium songaricum, Astragalus membranaceus, and sterile water were mixed, with a mass ratio of Cynomorium songaricum to sterile water of 1:22. The mixture was sterilized at 121 degrees Celsius for 15 minutes to obtain the fermentation substrate. Then, the obtained fermentation substrate was mixed with lactic acid bacteria (Lactobacillus rhamnosus ATCC7596 and Bifidobacterium longum CICC 6188 in a mass ratio of 10:11, with a mass ratio of fermentation substrate to lactic acid bacteria of 100:7. The mixture was fermented at 37 degrees Celsius for 72 hours, pre-frozen at -35 degrees Celsius for 2 hours, and freeze-dried at -25 degrees Celsius for 15 hours to obtain the fermentation powder.

[0048] (2) The kidneys are dried by roasting them over firewood and tiles in the traditional way and then ground into powder to obtain kidney powder.

[0049] (3) Grind and mix the fermented powder and kidney powder to obtain a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney.

[0050] Example 2: This embodiment prepares a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney, with the following components: 30g mulberry extract, 25g desert cistanche, 20g cynomorium, 20g dog kidney, and 5g astragalus. The mulberry extract is prepared as follows: Mulberry, ethanol, deionized water, and acetic acid are mixed at a mass ratio of 100:26 and 100:220, and extracted in a 56°C water bath for 1.5 hours under acidic conditions (pH 5.8). The mixture is then filtered to obtain the mulberry extract. The desert cistanche is *Cistanche deserticola* from Alashan, Inner Mongolia; the cynomorium is *Cistanche cynomorium* from Alashan, Inner Mongolia; and the astragalus is *Astragalus membranaceus* from Hunyuan, Shanxi.

[0051] This embodiment describes the preparation of a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys. The specific steps are as follows: (1) Before use, Cistanche deserticola, Cynomorium songaricum, and Astragalus membranaceus were ground into powder. Then, mulberry extract, Cistanche deserticola, Cynomorium songaricum, Astragalus membranaceus, and sterile water were mixed, with a mass ratio of Cynomorium songaricum to sterile water of 1:24. The mixture was sterilized at 121 degrees Celsius for 15 minutes to obtain a fermentation substrate. Then, the obtained fermentation substrate was mixed with lactic acid bacteria (Lactobacillus rhamnosus ATCC7596 and Bifidobacterium longum CICC 6188 in a mass ratio of 10:12, with a mass ratio of fermentation substrate to lactic acid bacteria of 100:5. The mixture was fermented at 37 degrees Celsius for 72 hours, pre-frozen at -35 degrees Celsius for 2 hours, and freeze-dried at -25 degrees Celsius for 15 hours to obtain fermentation powder.

[0052] (2) The kidneys are dried by roasting them over firewood and tiles in the traditional way and then ground into powder to obtain kidney powder.

[0053] (3) Grind and mix the fermented powder and kidney powder to obtain a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney.

[0054] Example 3: This embodiment prepares a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney, with the following components: 30g mulberry extract, 25g desert cistanche, 20g cynomorium, 20g sheep kidney, and 5g astragalus. The mulberry extract is prepared as follows: Mulberry, ethanol, deionized water, and acetic acid are mixed at a mass ratio of 100:24 (mulberry to ethanol) and 100:210 (mulberry to water). The mixture is then extracted in an acidic environment (pH 5.8) at 45°C for 1.8 hours, filtered, to obtain the mulberry extract. The desert cistanche is *Cistanche deserticola* from Alashan, Inner Mongolia; the cynomorium is *Cistanche cynomorium* from Alashan, Inner Mongolia; and the astragalus is *Astragalus membranaceus* from Hunyuan, Shanxi.

[0055] This embodiment describes the preparation of a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys. The specific steps are as follows: (1) Before use, Cistanche deserticola, Cynomorium songaricum, and Astragalus membranaceus were ground into powder. Then, mulberry extract, Cistanche deserticola, Cynomorium songaricum, Astragalus membranaceus, and sterile water were mixed, with a mass ratio of Cynomorium songaricum to sterile water of 1:22. The mixture was sterilized at 121 degrees Celsius for 15 minutes to obtain a fermentation substrate. Then, the obtained fermentation substrate was mixed with lactic acid bacteria (Lactobacillus rhamnosus ATCC7596 and Bifidobacterium longum CICC 6188 in a mass ratio of 10:10, with a mass ratio of fermentation substrate to lactic acid bacteria of 100:5. The mixture was fermented at 37 degrees Celsius for 78 hours, pre-frozen at -35 degrees Celsius for 2 hours, and freeze-dried at -25 degrees Celsius for 15 hours to obtain fermentation powder.

[0056] (2) The kidneys are dried by roasting them over firewood and tiles in the traditional way and then ground into powder to obtain kidney powder.

[0057] (3) Grind and mix the fermented powder and kidney powder to obtain a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney.

[0058] Example 4: This embodiment prepares a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys, with the following components: 30g mulberry extract, 25g desert cistanche, 20g cynomorium, 20g sheep kidney, and 5g astragalus. The mulberry extract is prepared as follows: Mulberry, ethanol, deionized water, and acetic acid are mixed at a mass ratio of 100:25 and 100:230, and extracted in a 52°C water bath for 1.5 hours under acidic conditions (pH 5.7). The mixture is then filtered to obtain the mulberry extract. The desert cistanche is *Cistanche deserticola* from Alashan, Inner Mongolia; the cynomorium is *Cistanche cynomorium* from Alashan, Inner Mongolia; and the astragalus is *Astragalus membranaceus* from Hunyuan, Shanxi.

[0059] This embodiment describes the preparation of a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys. The specific steps are as follows: (1) Before use, Cistanche deserticola, Cynomorium songaricum, and Astragalus membranaceus were ground into powder. Then, mulberry extract, Cistanche deserticola, Cynomorium songaricum, Astragalus membranaceus, and sterile water were mixed, with a mass ratio of Cynomorium songaricum to sterile water of 1:24. The mixture was sterilized at 121 degrees Celsius for 15 minutes to obtain a fermentation substrate. Then, the obtained fermentation substrate was mixed with lactic acid bacteria (Lactobacillus rhamnosus ATCC7596 and Bifidobacterium longum CICC 6188 in a mass ratio of 10:12, with a mass ratio of fermentation substrate to lactic acid bacteria of 100:6. The mixture was fermented at 37 degrees Celsius for 78 hours, pre-frozen at -35 degrees Celsius for 2 hours, and freeze-dried at -25 degrees Celsius for 15 hours to obtain fermentation powder.

[0060] (2) The kidneys are dried by roasting them over firewood and tiles in the traditional way and then ground into powder to obtain kidney powder.

[0061] (3) Grind and mix the fermented powder and kidney powder to obtain a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney.

[0062] Example 5: This embodiment prepares a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys, with the following components: 30g mulberry extract, 25g desert cistanche, 20g cynomorium, 20g sheep kidney, and 5g astragalus. The mulberry extract is prepared as follows: Mulberry, ethanol, deionized water, and acetic acid are mixed at a mass ratio of 100:25 (mulberry to ethanol) and 100:225 (mulberry to water). The mixture is then extracted in an acidic environment (pH 5.8) at 51°C for 1.6 hours, filtered, to obtain the mulberry extract. The desert cistanche is from Alashan, Inner Mongolia; the cynomorium is from Alashan, Inner Mongolia; and the astragalus is from Hunyuan, Shanxi.

[0063] This embodiment describes the preparation of a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidneys. The specific steps are as follows: (1) Before use, Cistanche deserticola, Cynomorium songaricum, and Astragalus membranaceus were ground into powder. Then, mulberry extract, Cistanche deserticola, Cynomorium songaricum, Astragalus membranaceus, and sterile water were mixed, with a mass ratio of Cynomorium songaricum to sterile water of 1:25. The mixture was sterilized at 121 degrees Celsius for 15 minutes to obtain a fermentation substrate. Then, the obtained fermentation substrate was mixed with lactic acid bacteria (Lactobacillus rhamnosus ATCC7596 and Bifidobacterium longum CICC 6188 in a mass ratio of 10:9, with a mass ratio of fermentation substrate to lactic acid bacteria of 100:6. The mixture was fermented at 37 degrees Celsius for 78 hours, pre-frozen at -35 degrees Celsius for 2 hours, and freeze-dried at -25 degrees Celsius for 15 hours to obtain fermentation powder.

[0064] (2) The kidneys are dried by roasting them over firewood and tiles in the traditional way and then ground into powder to obtain kidney powder.

[0065] (3) Grind and mix the fermented powder and kidney powder to obtain a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney.

[0066] Comparative Example 1: The preparation method of this comparative example is the same as that of Example 5, except that the mulberry extract is replaced with an equal mass of mulberry powder.

[0067] Comparative Example 2: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 35g of mulberry extract, 25g of desert cistanche, 20g of cynomorium, 20g of sheep kidney and 5g of astragalus.

[0068] Comparative Example 3: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 20g of desert cistanche, 20g of cynomorium, 20g of sheep kidney and 5g of astragalus.

[0069] Comparative Example 4: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 25g of desert cistanche, 23g of cynomorium, 20g of sheep kidney and 5g of astragalus.

[0070] Comparative Example 5: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 25g of desert cistanche, 20g of cynomorium, 15g of sheep kidney and 5g of astragalus.

[0071] Comparative Example 6: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 25g of desert cistanche, 20g of cynomorium, 20g of sheep kidney and 8g of astragalus.

[0072] Comparative Example 7: The preparation method of this comparative example is the same as that of Example 5, except that Astragalus membranaceus is omitted.

[0073] Comparative Example 8: The preparation method of this comparative example is the same as that of Example 5, except that Cynomorium songaricum is omitted.

[0074] Comparative Example 9: The preparation method of this comparative example is the same as that of Example 5, except that desert Cistanche deserticola is omitted.

[0075] Comparative Example 10: The preparation method of this comparative example is the same as that of Example 5, except that 3g of Epimedium is added.

[0076] Comparative Example 11: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 25g of desert cistanche, 20g of cynomorium, 20g of sheep kidney and 6g of astragalus.

[0077] Comparative Example 12: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 25g of desert cistanche, 20g of cynomorium, 20g of sheep kidney and 6.5g of astragalus.

[0078] Comparative Example 13: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 25g of desert cistanche, 20g of cynomorium, 20g of sheep kidney and 7g of astragalus.

[0079] Comparative Example 14: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 30g of mulberry extract, 25g of desert cistanche, 20g of cynomorium, 20g of sheep kidney and 7.5g of astragalus.

[0080] Comparative Example 15: The preparation method of this comparative example is the same as that of Example 5, except that the components are as follows: 25g of desert Cistanche deserticola, 20g of Cynomorium songaricum and 5g of Astragalus membranaceus.

[0081] Comparative Example 16: The preparation method of this comparative example is the same as that of Example 5, except that: (1) Before use, Cistanche deserticola, Cynomorium songaricum and Astragalus membranaceus were ground into powder separately. Then, mulberry extract, Cistanche deserticola, Cynomorium songaricum and Astragalus membranaceus were mixed to obtain a mixed powder.

[0082] (2) The kidneys are dried by roasting them over firewood and tiles in the traditional way and then ground into powder to obtain kidney powder.

[0083] (3) Grind and mix the mixed powder and kidney powder to obtain a compound preparation of mulberry, cistanche and cynomorium for tonifying the kidney.

[0084] Test Example 1: The efficacy of the mulberry, cistanche, and cynomorium compound preparations for tonifying the kidneys in Examples 1-5 and Comparative Examples 1-16 was tested. The testing method was as follows: Four-week-old SD rats, half male and half female, were randomly divided into a model control group and a model group. The model group was administered hydrocortisone solution (50 mg / kg) by gavage once daily, with free access to water and food, for 10 consecutive days until modeling was successful. The model control group was given an equal volume of physiological saline daily for 10 consecutive days. Thirty rats were randomly selected from the model control group and 660 rats from the model group. The model group rats were randomly divided into 22 subgroups, which were administered the mulberry, cistanche, and cynomorium compound preparations for tonifying the kidneys in Examples 1-5 and Comparative Examples 1-16, and physiological saline (blank control group), respectively, twice daily (morning and evening), 300 mg / kg each time, for 30 consecutive days. The model control group rats received no treatment and were fed normally. The physiological state of the rats was observed (constant food intake and body weight mean that the change rate of each is no greater than 3.5%), and the changes in serum testosterone levels were recorded. The test results are shown in Tables 1-2 and 2. Figure 1 As shown.

[0085] Table 1. Efficacy of Examples 1-5 and Comparative Examples 1-16:

[0086] Table 2. Testosterone levels in Examples 1-5 and Comparative Examples 1-16:

[0087] In Table 2, 'a' indicates that compared with the model control group, P < 0.05.

[0088] According to Tables 1-2 and Figure 1It can be seen that the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney provided by this invention can effectively alleviate the symptoms of kidney deficiency in rats and promote the increase of testosterone levels. As shown in Comparative Examples 1-16, the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney provided by this invention, through the combination of five medicinal materials and strict control of the dosage, achieves a good warming and tonifying effect on the kidney. The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney provided by this invention relies on both the combination and dosage of medicinal materials, and the combined fermentation of *Lactobacillus rhamnosus* ATCC7596 and *Bifidobacterium longum* CICC6188; both are indispensable. The purpose of this invention, employing fermentation, is to efficiently extract the active ingredients from a combination of medicinal herbs, ultimately yielding a mulberry, cistanche, and cynomorium compound preparation for kidney tonification. However, during the aforementioned rat experiments, it was found that the drug combination protected by this invention exhibited superior performance compared to other herbal combinations. This may be because new compounds were generated during fermentation, synergistically interacting with the herbal combination for a better effect; or fermentation may have resulted in a higher absolute amount of active ingredients in the herbal combination compared to unfermented herbs, making it equivalent to a higher dosage of traditional herbal formulas. Furthermore, the varying degrees of fermentation among different herbs may have altered the proportions of the active ingredients actually exerting their effects, potentially causing adverse reactions to the microbial community in the combination, thus preventing other herbal combinations with adjusted dosages from showing better efficacy. The specific mechanism is still under further investigation.

[0089] Test Example 2: Forty-eight patients with similar degrees of kidney deficiency and poor spleen and stomach function were recruited. They were randomly divided into 16 groups and clinically administered the mulberry, cistanche, and cynomorium compound preparations used for tonifying the kidneys in Examples 1-5 and Comparative Examples 1-10 and Comparative Example 16, respectively. The dosage was 20 grams twice daily, morning and evening, taken with 60°C warm water, for 30 consecutive days. The results are shown in Table 3. Further testing was conducted on Example 5 with 85 participants, and the results are shown in Table 4.

[0090] Table 3 Test results of Examples 1-5, Comparative Examples 1-10, and Comparative Example 16:

[0091] Table 4 Test results of Example 5:

[0092] In Table 4, if the tongue or pulse condition improves and appetite, abdominal distension, or indigestion does not worsen (improves or remains unchanged), it is recorded as a significant improvement; if the tongue or pulse condition improves and appetite, abdominal distension, or indigestion worsens, or if the tongue or pulse condition remains unchanged and appetite, abdominal distension, or indigestion improves, it is recorded as a slight improvement; if the tongue or pulse condition remains unchanged and appetite, abdominal distension, or indigestion remains unchanged, it is recorded as no worsening; if the tongue or pulse condition remains unchanged and appetite, abdominal distension, or indigestion worsens, or if the tongue or pulse condition worsens but appetite, abdominal distension, or indigestion does not worsen, it is recorded as a worsening; if the tongue or pulse condition worsens and appetite, abdominal distension, or indigestion worsens, it is recorded as a significant worsening.

[0093] As can be seen from Tables 3 and 4, the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidneys provided by this invention can effectively reduce the degree of kidney deficiency in patients, gently nourish and tonify the kidneys, and has no obvious side effects on spleen and stomach function. It is suitable for patients with kidney deficiency and poor spleen and stomach function, and solves the problem that patients have insufficient spleen and stomach function and that the spleen and stomach function further declines during the medication process, resulting in poor kidney tonifying effect of the drug and inability to take the drug for a long time.

[0094] The embodiments of the present invention have been described above; however, these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. All other embodiments obtained by those skilled in the art based on the above embodiments of the present invention without inventive effort are within the protection scope of the present invention.

Claims

1. A compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney, characterized in that, The product comprises the following components: 30 wt% mulberry extract, 25 wt% desert cistanche, 20 wt% cynomorium, 20 wt% kidney extract, and 5 wt% astragalus. The kidneys are sheep kidneys or dog kidneys; The preparation method of the mulberry extract includes the following steps: mixing mulberry, ethanol, water and acid, extracting under acidic conditions, and then separating the solid and liquid components to obtain the mulberry extract; The preparation method of the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney includes the following steps: (1) Mulberry extract, desert cistanche, cynomorium, astragalus and sterile water are mixed and sterilized to obtain a fermentation substrate. Then the fermentation substrate is mixed with lactic acid bacteria and fermented and then freeze-dried to obtain fermentation powder. The lactic acid bacteria are Lactobacillus rhamnosus and Bifidobacterium longum; The mass ratio of Lactobacillus rhamnosus to Bifidobacterium longum is 10:8~12; (2) The kidneys are dried by baking with firewood and tiles and then ground into powder to obtain kidney powder; (3) The fermented powder and kidney powder are mixed to obtain the mulberry, cistanche and cynomorium compound preparation for tonifying the kidney; There is no requirement for the time order of steps (1) and (2).

2. The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney according to claim 1, characterized in that, The mass ratio of mulberry to ethanol is 100:23~28; The mass ratio of mulberries to water is 100:200~240.

3. The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney according to claim 1, characterized in that, The pH value of the acidic conditions is 5.5~6.

4. The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney according to claim 1, characterized in that, The extraction temperature is 45-56 degrees Celsius, and the holding time is 1.5-1.8 hours. The extraction is carried out in a water bath.

5. The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney according to claim 1, characterized in that, The Astragalus mentioned is the Astragalus membranaceus from the north.

6. A method for preparing a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney, characterized in that, The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney is the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney as described in any one of claims 1 to 5, comprising the following steps: (1) Mulberry extract, desert cistanche, cynomorium, astragalus and sterile water are mixed and sterilized to obtain a fermentation substrate. Then the fermentation substrate is mixed with lactic acid bacteria and fermented and then freeze-dried to obtain fermentation powder. The lactic acid bacteria are Lactobacillus rhamnosus and Bifidobacterium longum; The mass ratio of Lactobacillus rhamnosus to Bifidobacterium longum is 10:8~12; (2) The kidneys are dried by baking with firewood and tiles and then ground into powder to obtain kidney powder; (3) The fermented powder and kidney powder are mixed to obtain the mulberry, cistanche and cynomorium compound preparation for tonifying the kidney; There is no requirement for the time order of steps (1) and (2).

7. The application of a compound preparation of mulberry, cistanche, and cynomorium for tonifying the kidney in the preparation of kidney-tonifying drugs, characterized in that, The mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney is the mulberry, cistanche, and cynomorium compound preparation for tonifying the kidney as described in any one of claims 1 to 5.

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