CN116870053A - Medicine for improving sexual function and preparation method thereof - Google Patents
Medicine for improving sexual function and preparation method thereof Download PDFInfo
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- CN116870053A CN116870053A CN202310732234.4A CN202310732234A CN116870053A CN 116870053 A CN116870053 A CN 116870053A CN 202310732234 A CN202310732234 A CN 202310732234A CN 116870053 A CN116870053 A CN 116870053A
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/64—Orobanchaceae (Broom-rape family)
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/08—Drugs for genital or sexual disorders; Contraceptives for gonadal disorders or for enhancing fertility, e.g. inducers of ovulation or of spermatogenesis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
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- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Diabetes (AREA)
- Public Health (AREA)
- Medicinal Chemistry (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- General Chemical & Material Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Biotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Reproductive Health (AREA)
- Mycology (AREA)
- Endocrinology (AREA)
- Microbiology (AREA)
- Botany (AREA)
- Pregnancy & Childbirth (AREA)
- Gynecology & Obstetrics (AREA)
- Alternative & Traditional Medicine (AREA)
- Emergency Medicine (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Medical Informatics (AREA)
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- General Engineering & Computer Science (AREA)
- Genetics & Genomics (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The application relates to the technical field of biological fermentation, in particular to a medicine for improving sexual function and a preparation method thereof. The medicament for improving the sexual function contains cistanche deserticola fermentation products, can treat male erectile dysfunction caused by diabetes mellitus, has better effect than unfermented cistanche deserticola extract, and has application potential in improving sexual function hair.
Description
Technical Field
The application relates to the technical field of biological fermentation, in particular to a medicine for improving sexual function and a preparation method thereof.
Background
Cistanche deserticola has aliases such as golden bamboo shoots and ground essence, is a perennial plant and is mainly distributed in the northwest desert area of China. It is warm in nature, sweet and salty. The stems of cistanche deserticola were first recorded in Shennong's traditional Chinese medicine. Cistanche has long been used as a traditional herbal medicine for treating kidney deficiency, impotence, senile constipation, soreness and weakness of waist and knees and blood deficiency, and pharmacological studies have shown that extracts of cistanche have various biological functions including regulating immune activity, enhancing sexual function, promoting hair growth and other benefits of antioxidant, liver protecting, anti-osteoporosis activity and anti-tumor. In addition, in vitro studies have shown that cistanche extract can protect dopamine-like neurons from induced oxidative damage and significantly increase the levels of nerve growth factor and brain-derived neurotrophic factor. Cistanche deserticola also has a certain anti-aging effect. Research shows that aging can be classified into the characteristics of reduced physiological functions, reduced memory, reduced immune functions and the like. The cistanche deserticola feed is fed to the drosophila melanogaster, so that the resistance of the drosophila melanogaster to oxidative stress is improved, and the life of the fed drosophila melanogaster is prolonged to a certain extent.
There are various ways of fermenting, but the most effective method is to ferment with microorganisms. As the microorganism can produce various enzymes to catalyze the traditional Chinese medicine, the active ingredients, the application mode and the medicine property of the traditional Chinese medicine can be changed to a certain extent. The fermentation is carried out by utilizing different strains, the produced results are different, and the proper strains can be beneficial to enhancing the treatment effect of the medicine and reducing the toxic and side effects, provide a new direction for producing new medicine sources and further develop the dosage form of the traditional Chinese medicine preparation, and can be used in clinic with better safety.
Microbial fermentation of traditional Chinese medicine is usually a natural fermentation mode by mixing multiple strains. The traditional fermentation method has a certain limit, the traditional process does not have advanced processing technology, and the quality and the drug effect of the drug can be greatly different due to a little improper operation.
Disclosure of Invention
The application provides a medicine for improving sexual function, which has controllable production conditions, good product stability and application potential in the aspect of improving sexual function.
The application is realized by the following modes:
in a first aspect, the application provides a medicament for improving sexual function, wherein the medicament for improving sexual function is a medicament for treating male erectile dysfunction caused by diabetes, the medicament for improving sexual function contains a cistanche fermentation product, the cistanche fermentation product is obtained by fermenting cistanche with lactobacillus rhamnosus Y5, and the lactobacillus rhamnosus Y5 has a preservation number of CGMCC No.24886.
Preferably, the drug for improving sexual function is a drug for treating male erectile dysfunction caused by diabetes by improving internal pressure of corpora cavernosa.
The medicine, preferably cistanche deserticola ferment, is obtained by inoculating lactobacillus rhamnosus Y5 into a fermentation culture solution containing cistanche deserticola for culture to obtain a fermentation solution, and extracting the fermentation solution by heating and refluxing.
The medicine preferably contains 1-2wt% of glucose and 10-15wt% of cistanche deserticola in fermentation culture solution.
The medicament, preferably the medicament for improving sexual function, comprises a pharmaceutically acceptable carrier.
The medicine is preferably cultured by a shaking table, and is fermented for 6d at 35 ℃, and the rotating speed of the shaking table is 160r/min.
The medicine is preferably specifically operated by inoculating seed liquid into fermentation culture liquid, fermenting for 6d at 35 ℃ by a shaking table, and obtaining fermentation liquid at the rotating speed of 160 r/min; the volume ratio of the seed liquid to the fermentation culture liquid is 1:50, and the seed liquid contains 10 8 cfu/mL of Lactobacillus rhamnosus.
In a second aspect, the application provides an application of cistanche deserticola in preparing a medicament and/or a medicament intermediate for treating male erectile dysfunction caused by diabetes, wherein cistanche deserticola is fermented by lactobacillus rhamnosus Y5 to obtain a cistanche deserticola fermentation product, the cistanche deserticola fermentation product is used for preparing the medicament and/or the medicament intermediate for treating male erectile dysfunction caused by diabetes, and the lactobacillus rhamnosus Y5 is stored in a CGMCC No.24886.
Preferably, the medicament and/or the medicament intermediate is a medicament and/or medicament intermediate for treating male erectile dysfunction caused by diabetes mellitus by improving internal pressure of the corpora cavernosa.
In a third aspect, the application provides an application of lactobacillus rhamnosus Y5 in preparing a medicament and/or a medicament intermediate for treating male erectile dysfunction caused by diabetes, wherein the lactobacillus rhamnosus Y5 has a preservation number of CGMCC No.24886, cistanche fermentation is obtained by fermenting cistanche with lactobacillus rhamnosus Y5, and the cistanche fermentation is used for preparing the medicament and/or the medicament intermediate for treating male erectile dysfunction caused by diabetes.
The application has the beneficial effects that:
the cistanche deserticola fermented product obtained by the modern fermentation process has controllable production conditions and good product stability, can obviously improve the internal pressure ICP of the corpora cavernosa of the diabetic rat, has better effect of improving sexual function than the unfermented cistanche deserticola extract, and has application potential in preparing medicines for improving sexual function.
Information on preservation of strains
Preservation time: 2022, 5 and 12 days,
preservation unit: the China general microbiological culture Collection center,
preservation number: CGMCC No.24886,
deposit unit address: beijing city, chaoyang area, north Chenxi Lu 1 hospital the institute of microbiology of the national academy of sciences 3,
classification naming: lactobacillus rhamnosus Lactobacillus rhamnosus.
Drawings
Figure 1 shows the cell viability under the action of the fermented cistanche extract at different concentrations,
fig. 2 is a graph showing changes in intracavernosal pressure in rats treated differently for diabetic erectile dysfunction.
Detailed Description
A medicament for improving sexual function is a medicament for treating male erectile dysfunction caused by diabetes, and comprises cistanche deserticola fermentation product, wherein the cistanche deserticola fermentation product is obtained by fermenting cistanche deserticola with lactobacillus rhamnosus Y5, and the lactobacillus rhamnosus Y5 has a preservation number of CGMCC No.24886.
The cistanche powder or the cistanche block can be used, and in order to make the fermentation effect better, the cistanche powder is preferably used, and the washed and dried cistanche is ground into powder by a mechanical grinder.
The medicine for improving sexual function treats male erectile dysfunction caused by diabetes by improving internal pressure of corpora cavernosa.
The cistanche fermentation product is obtained by inoculating lactobacillus rhamnosus Y5 into a fermentation culture solution containing cistanche for culture to obtain a fermentation solution, and extracting the fermentation solution by heating and refluxing. The extraction temperature by heating and refluxing is 100 ℃.
The fermentation culture solution contains 1-2wt% glucose and 10-15wt% cistanche deserticola.
The medicine for improving sexual function contains a pharmaceutically acceptable carrier.
The culture is shake culture, fermentation is carried out for 6d at 35 ℃, and the rotation speed of the shake culture is 160r/min.
Inoculating the seed liquid into a fermentation culture solution, and fermenting for 6d at 35 ℃ by a shaking table, wherein the rotation speed of the shaking table is 160r/min to obtain a fermentation solution; the volume ratio of the seed liquid to the fermentation culture liquid is 1:50, and the seed liquid contains 10 8 cfu/mL of Lactobacillus rhamnosus.
An application of cistanche in preparing the medicines and/or intermediates for treating the erectile dysfunction caused by diabetes is disclosed, which features that the lactobacillus rhamnosus Y5 is used to ferment to obtain the fermented cistanche, which is used to prepare the medicines and/or intermediates for treating the erectile dysfunction caused by diabetes, and the lactobacillus rhamnosus Y5 has the preservation number of CGMCC No.24886.
In a third aspect, the application provides an application of lactobacillus rhamnosus Y5 in preparing a medicament and/or a medicament intermediate for treating male erectile dysfunction caused by diabetes, wherein the lactobacillus rhamnosus Y5 has a preservation number of CGMCC No.24886, cistanche fermentation is obtained by fermenting cistanche with lactobacillus rhamnosus Y5, and the cistanche fermentation is used for preparing the medicament and/or the medicament intermediate for treating male erectile dysfunction caused by diabetes.
The lactobacillus rhamnosus Y5 used in the following examples is obtained by screening from the purchased lactobacillus rhamnosus powder by the applicant. Lactobacillus rhamnosus Y5 has been deposited in the China general microbiological culture Collection center, accession number: CGMCC No.24886. Lactobacillus plantarum L1 and Lactobacillus casei L3 are commercially available Lactobacillus plantarum and Lactobacillus casei, and because a plurality of strains of Lactobacillus plantarum and Lactobacillus casei are used in the experimental process, they are designated as Lactobacillus plantarum L1 and Lactobacillus casei L3 for the sake of convenience of distinction. All strains used in the experimental procedure, for the sake of brevity, are not all those mentioned in this embodiment, and only the 3 strains having the best effect among them are selected.
Example 1
The cistanche fermentation product is prepared according to the following steps:
(1) Each liter of seed culture medium comprises 5g of yeast extract, 10g of tryptone, 10g of sodium chloride and 1000mL of distilled water, lactobacillus rhamnosus Y5 is inoculated into the seed culture medium, and shake culture is carried out at 37 ℃ and 160r/min to obtain lactobacillus rhamnosus Y5 with the content of 10 8 cfu/mL of seed solution;
(2) Each liter of fermentation culture solution comprises 10g of glucose, 100g of cistanche deserticola and 1000mL of water, the fermentation culture solution is sterilized for 20min at 121 ℃ in a high-pressure steam sterilization pot before use, 150mL of fermentation culture solution is placed in a 500mL shaking bottle, 3mL of seed solution is inoculated, the fermentation is carried out for 6d at 35 ℃ of a shaking table, and the rotation speed of the shaking table is 160r/min, so that fermentation solution is obtained;
(3) Reflux fermenting the fermentation liquor for 1h at 100deg.C, filtering and centrifuging to obtain herba cistanches fermented product, named as Y5 herba cistanches fermented product (FCD).
Example 2
In comparison with example 1, lactobacillus rhamnosus Y5 was replaced with lactobacillus plantarum L1, and the rest of the procedure was the same as in example 1, to obtain a cistanche deserticola ferment, designated as L1 cistanche deserticola Ferment (FCD).
Example 3
In comparison with example 1, lactobacillus rhamnosus Y5 was replaced with lactobacillus casei L3, and the rest of the procedure was the same as in example 1, to obtain a cistanche fermentation product, designated as L3 cistanche fermentation product (FCD).
Example 4
In comparison with example 1, 3mL of the seed solution was replaced with 3mL of sterile water, and the above steps (2) - (3) were repeated to obtain cistanche extract (CD).
Polysaccharide and phenethyl alcohol glycoside content determination
Detecting the content of polysaccharide and total phenethyl alcohol glycoside in the cistanche fermentation product or cistanche extract prepared in the above examples 1-4 by ultraviolet spectrophotometry.
The polysaccharide content of the cistanche fermentation product or the cistanche extract in examples 1-4 was measured at a wavelength of 260nm, and the phenylethanoid glycoside content of the cistanche fermentation product or the cistanche extract in examples 1-4 was measured at a wavelength of 350 nm. The results are shown in Table 1 below.
TABLE 1 polysaccharide and phenethyl alcohol glycoside content of cistanche fermentation or cistanche extract of examples 1-4
| Product(s) | Polysaccharide | Total phenethyl alcohol glycoside | |
| Example 1 | Y5 cistanche salsa fermentation extract (FCD) | 12.4％ | 3.0％ |
| Example 2 | L1 cistanche deserticola fermentation extract (FCD) | 11.4％ | 2.6％ |
| Example 3 | L3 cistanche deserticola fermentation extract (FCD) | 9.4％ | 2.3％ |
| Example 4 | Cistanche extract (NCD) | 9.8％ | 2.4％ |
As can be seen from the above table, the polysaccharides and phenethyl alcohol glycosides in the fermented extracts (FCD) of Y5 herba cistanches and L1 herba cistanches are obviously improved compared with those in the fermented extracts (NCD), and the polysaccharides and phenethyl alcohol glycosides in the fermented extracts (FCD) of Y5 herba cistanches are obviously improved compared with those in the fermented extracts (FCD) of L1 herba cistanches.
Cell and animal test
1. Y5 cistanche deserticola fermentation extract (FCD) to H 2 O 2 Protection of induced RSC-96 cells from apoptosis
Mo Xibao (RSC-96) was applied in accordance with 3X 10 5 Density of individual/mL (100. Mu.L/well) was seeded into 96-well plates, 37C 5% CO 2 Cells were cultured in a constant temperature incubator for 24 hours. Setting blank control group, control group and experimental group (fermented product with different concentrations), pretreating cells with extract with different concentrations for 6 hr, adding 50 μm H into the control group and experimental group 2 O 2 Cells were stimulated for 12 hours. Subsequently, 10. Mu.L of MTT solution was added to each well, incubated at 37℃for 4 hours, the medium was removed, 150. Mu.L of LDMSO was added to dissolve formaldehyde crystals, and the Optical Density (OD) was measured at 490 nm.
The results are shown in FIG. 1, wherein the blank is a blank control group, the hydrogen peroxide is a control group, and 1, 10, 50 and 100 are respectively prepared from fermented extract of cistanche deserticola containing Y5FCD) 1 μg/mL, 10 μg/mL, 50 μg/mL, 100 μg/mL, data indicating that Y5 cistanche fermentation extract (FCD) vs. H 2 O 2 The induced schwann cells have certain protection effect, and the protection effect is best when the concentration is 10 ug/mL.
2. Effects of cistanche fermentation on erectile function of diabetic rats
All experimental procedures were approved by the Shandong university animal protection and use Committee.
Establishment of diabetic ED rat model:
48 male Sprague-Dawley rats of 8 weeks of age were selected. Animals were kept in a Specific Pathogen Free (SPF) environment at 22-24 ℃ for 12 hours light/12 hours dark cycles per week, routinely given water and food. To establish a diabetic rat model, 40 rats were intraperitoneally injected with streptozotocin after fasting, and the remaining 8 rats were given an equal volume of 0.1mol/L citrate phosphate buffer (ph=4.2) subcutaneously. After 72h of monitoring, rats with blood glucose levels exceeding 16.7mM were identified as diabetic.
After 12 weeks, the apomorphine apomopine test was used to identify the presence of DMED by observing penile congestion, length and circumference changes. 40 DMED rats were divided into 5 groups by APO test and treated with different treatment regimens, which were enema, and specific groupings and treatments were as follows:
DM group: n=8, blank, no treatment;
dm+lncd group: n=8, treatment with 100mg/kg cistanche extract (NCD);
dm+hncd group: n=8, treatment with 300mg/kg cistanche extract (NCD);
dm+lfcd group: n=8, treatment with 100mg/kg Y5 cistanche deserticola fermented extract (FCD);
dm+hfcd group: n=8, treatment with 300mg/kg Y5 cistanche deserticola fermented extract (FCD).
Evaluation of erectile function:
after 8 weeks of treatment, intracavernosal pressure (ICP) values and mean systemic arterial pressure (MAP) were measured in different groups of male rats. After anesthesia (intraperitoneal, 70 mg/kg) with 2.5% sodium pentobarbital, male rats were placed face up. The left carotid artery was cannulated with a PE-50 tube and the systemic arterial blood pressure was recorded with a pressure sensor. A needle (23 gauge) filled with heparin solution (250U/mL) was connected to the sensor and inserted into the sponge to record ICP. The cavernous nerve was isolated and stimulated with a hook-shaped bipolar electrode at 5V at 25Hz for 60s with a pulse width of 5ms. Changes in ICP and MAP were recorded by BL-420V pressure sensor system and the data was visualized using software.
As shown in fig. 2, the internal penile cavernous pressure (ICP) values of dm+lfcd group and dm+hfcd group were significantly increased compared to dm+lncd group, dm+hncd group, wherein dm+hfcd group increase effect most clearly demonstrated that Y5 cistanche fermented extract (FCD) treatment could improve ICP level in diabetic rats, and Y5 cistanche fermented extract (FCD) improvement effect was significantly superior to unfermented cistanche extract (NCD).
Claims (10)
1. The medicine for improving sexual function is characterized in that the medicine for improving sexual function is used for treating male erectile dysfunction caused by diabetes, the medicine for improving sexual function contains cistanche deserticola fermentation product, the cistanche deserticola fermentation product is obtained by fermenting cistanche deserticola with lactobacillus rhamnosus Y5, and the lactobacillus rhamnosus Y5 has a preservation number of CGMCC No.24886.
2. The medicament according to claim 1, wherein the medicament for improving sexual function is a medicament for treating male erectile dysfunction caused by diabetes by improving internal pressure of corpora cavernosa.
3. The medicament according to claim 1 or 2, characterized in that the cistanche deserticola ferment is obtained by inoculating lactobacillus rhamnosus Y5 into a fermentation broth containing cistanche deserticola for culture to obtain a fermentation broth, and extracting the fermentation broth by heating and refluxing.
4. A medicament according to claim 3, characterized in that the fermentation broth contains 1-2wt% glucose and 10-15wt% cistanche deserticola.
5. The medicament according to claim 2, characterized in that the medicament for improving sexual function comprises a pharmaceutically acceptable carrier.
6. A medicament according to claim 3, characterized in that the cultivation is shaking cultivation, fermentation is carried out for 6d at 35 ℃, and the rotation speed of the shaking cultivation is 160r/min.
7. A medicament according to claim 3, characterized in that the specific operation is to inoculate seed liquid into fermentation broth, ferment for 6d at 35 ℃ with rotation speed of 160r/min, to obtain fermentation broth; the volume ratio of the seed liquid to the fermentation culture liquid is 1:50, and the seed liquid contains 10 8 cfu/mL of Lactobacillus rhamnosus Y5.
8. The application of cistanche deserticola in preparing a medicament and/or a medicament intermediate for treating male erectile dysfunction caused by diabetes is characterized in that cistanche deserticola is fermented by lactobacillus rhamnosus Y5 to obtain a cistanche deserticola fermentation product, the cistanche deserticola fermentation product is used for preparing the medicament and/or the medicament intermediate for treating male erectile dysfunction caused by diabetes, and the preservation number of lactobacillus rhamnosus Y5 is CGMCC No.24886.
9. Use according to claim 8, characterized in that the medicament and/or the pharmaceutical intermediate is a medicament and/or pharmaceutical intermediate for the treatment of diabetes induced male erectile dysfunction by improving the internal pressure of the corpora cavernosa.
10. The application of lactobacillus rhamnosus Y5 in preparing a medicament and/or a medicament intermediate for treating male erectile dysfunction caused by diabetes is characterized in that the lactobacillus rhamnosus Y5 has a preservation number of CGMCC No.24886, cistanche fermentation is performed by using lactobacillus rhamnosus Y5 to ferment cistanche, and the cistanche fermentation is used for preparing the medicament and/or the medicament intermediate for treating male erectile dysfunction caused by diabetes.
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