Application of lactobacillus salivarius in preparation of product for improving male hormone secretion of old people
By using Lactobacillus salivarius PC114 strain to regulate the steroid hormone biosynthesis pathway, the problems of large side effects, short-term, unstable and high cost of treating the reduction of elderly androgens in the prior art were solved, and safe, effective and economical androgen secretion enhancement effect was achieved.
Patent Information
- Application Number
- CN202510035743.0
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-01-09
- Publication Date
- 2025-04-15
AI Technical Summary
The prior art methods used to treat the reduction of androgen in the elderly have problems of large side effects, short-term effects, unstable and high cost, and there is a lack of effective lactic acid bacteria resources to improve the secretion of androgen in the elderly.
The bacterial agent was prepared by Lactobacillus salivarius PC114 strain with deposit number CGMCC No. 23518, which was prepared by fermentation culture and mixed with skim milk, which significantly regulated the steroid hormone biosynthesis pathway and promoted the production of pregnenolone, 11β-hydroxyandrothenone and 19-oxoandrost-4-en-3,17-dione.
It significantly improves the secretion of androgens in elderly people, regulates the biosynthesis pathway of steroid hormones, alleviates health problems caused by androgen reduction, and provides a safe, effective and economical treatment plan.
Smart Images
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Abstract
Description
Technical Field
[0001] This specification relates to the field of microbial technology, and in particular to the use of Lactobacillus salivarius in the preparation of a product for enhancing male hormone secretion in the elderly. Background Art
[0002] Androgens are important hormones in the male reproductive system and are essential for maintaining libido, sexual function, the development of male characteristics (such as head and body hair), muscle mass, bone density, and general health. As men age, their androgen levels may gradually decline, a condition known as "generalized androgen deprivation," which can lead to a range of sexual health problems that not only affect quality of life but can also cause physical and mental health challenges.
[0003] There are many clinical methods for treating androgen deficiency. One of the most common treatments is androgen replacement therapy, which is usually used in the form of skin gel, oral medication, injection or implantable hormone preparations. However, this treatment is not suitable for everyone and may have the following risks: (1) large side effects, which can easily cause gastrointestinal discomfort and loss of appetite; (2) short-lasting effects. Usually, after taking the medicine, it is necessary to continue taking it for several courses to maintain the therapeutic effect, and the effect is less sustainable; (3) unstable effects. The treatment mechanism is not clear enough, so the treatment effect often varies from person to person and is not stable; (4) high cost: long-term use of drugs will bring great financial pressure to patients.
[0004] Lactic acid bacteria are a special type of microbial flora found in the human body. They typically possess a variety of beneficial functions, such as promoting nutrient absorption, synthesizing essential vitamins, and regulating immunity. With the rapid advancement of microbial research, while we have a deep understanding of the connection between microbial flora and specific human diseases, there are currently few clinically available lactic acid bacteria for enhancing androgen secretion in the elderly. Therefore, the search for lactic acid bacteria that can promote androgen secretion in the elderly, thereby alleviating a series of health problems caused by androgen decline, has become an important breakthrough in this field. Summary of the Invention
[0005] In order to solve the above technical problems, the present application provides an application of Lactobacillus salivarius in preparing a product for enhancing male hormone secretion in the elderly, wherein the Lactobacillus salivarius is the Lactobacillus salivarius PC114 strain with a preservation number of CGMCC No. 23518.
[0006] The beneficial effects of this application include but are not limited to:
[0007] The present invention is the first to use Lactobacillus salivarius strain PC114 (CGMCC NO.23518) to enhance male hormone secretion in the elderly. It is the first to use Lactobacillus salivarius strain PC114 (CGMCC NO.23518) to prepare a bacterial agent. It reveals that Lactobacillus salivarius strain PC114 (CGMCC NO.23518) has a new use in promoting the levels of pregnenolone, 11β-hydroxyandrostenedione, and 19-oxoandrost-4-ene-3,17-dione, and significantly regulating the body's steroid hormone biosynthesis pathway. Lactobacillus salivarius strain PC114 (CGMCC NO.23518) has broad application prospects in products that alleviate a series of health problems caused by decreased androgen levels. BRIEF DESCRIPTION OF THE DRAWINGS
[0008] Figure 1 Shown are the major regulated pathways intervened by Lactobacillus salivarius strain PC114 (CGMCC NO. 23518);
[0009] Figure 2 Shown is the effect of Lactobacillus salivarius strain PC114 (CGMCC NO.23518) intervention on pregnenolone content;
[0010] Figure 3 Shown is the effect of Lactobacillus salivarius strain PC114 (CGMCC NO.23518) intervention on 11β-hydroxyandrostenedione content;
[0011] Figure 4 Shown is the effect of intervention with Lactobacillus salivarius strain PC114 (CGMCC NO.23518) on the content of 19-oxoandrost-4-ene-3,17-dione. DETAILED DESCRIPTION
[0012] As used in this specification and claims, unless the context clearly indicates otherwise, the words "a," "an," "an," and / or "the" do not refer to the singular but also include the plural. Generally speaking, the terms "comprises" and "include" only indicate the inclusion of the steps and elements specifically identified, and these steps and elements do not constitute an exclusive list. A method or apparatus may also include other steps or elements.
[0013] The present invention discovered for the first time that the Lactobacillus salivarius PC114 strain has the effect of enhancing the secretion of male hormones in the elderly, can regulate the steroid biosynthesis signaling pathway, and promote the increase of the content of pregnenolone, 11β-hydroxyandrostenedione and 19-oxoandrost-4-ene-3,17-dione, and can be used in products that enhance the secretion of male hormones in the elderly.
[0014] The present application first provides an application of Lactobacillus salivarius in preparing a product for enhancing male hormone secretion in the elderly. The Lactobacillus salivarius is the Lactobacillus salivarius PC114 strain with a preservation number of CGMCC No. 23518.
[0015] In certain embodiments of the present invention, the product for enhancing androgen secretion in the elderly has one or more of the following functions:
[0016] 1) increasing the levels of androgens and their derivatives, preferably increasing the levels of pregnenolone, 11β-hydroxyandrostenedione and 19-oxoandrost-4-ene-3,17-dione;
[0017] 2) Regulate steroid biosynthesis signaling pathways.
[0018] In a preferred embodiment, the products include but are not limited to health products, food, medicines, etc.
[0019] In a preferred embodiment, the dosage form of the product includes injection, suspension, powder, tablet or granule.
[0020] In certain embodiments of the present invention, the product is a drug, which further comprises a pharmaceutically acceptable carrier or excipient.
[0021] "Pharmaceutically acceptable" means that the drugs do not produce adverse, allergic or other untoward reactions when properly administered to animals or humans.
[0022] "Pharmaceutically acceptable carriers or excipients" should be compatible with the active ingredient, that is, they can be mixed with it without significantly reducing the efficacy of the drug under normal circumstances. Specific examples of substances that can serve as pharmaceutically acceptable carriers or excipients include sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium methylcellulose, ethylcellulose, and methylcellulose; tragacanth powder; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil, and cocoa butter; polyols such as propylene glycol, glycerol, sorbitol, mannitol, and polyethylene glycol; alginic acid; emulsifiers such as Tween; wetting agents such as sodium lauryl sulfate; colorants; flavorings; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline solution; and phosphate buffer, etc. These materials are used as needed to aid in the stability of the formulation or to help increase the activity or its bioavailability or to produce an acceptable taste or flavor in the case of oral administration.
[0023] In some embodiments, the drug of the present application can be administered in a monotherapy (e.g., without concomitant administration of any additional therapeutic agent, or without concomitant administration of any additional treatment and against the same disease to be treated or prevented by the drug of the present application). In some embodiments, the drug of the present application can also be administered in combination with one or more other therapeutic agents or administered simultaneously.
[0024] In a preferred embodiment, the product may contain a Lactobacillus salivarius bacterial agent. Preferably, the bacterial agent includes the strain Lactobacillus salivarius PC114 with a deposit number of CGMCC No. 23518.
[0025] In a specific embodiment, the preparation method of the bacterial agent may include: 1) inoculating Lactobacillus salivarius strain PC114 into a liquid culture medium for fermentation and culture; 2) separating and collecting the bacterial cells cultured in step 1) and collecting the bacterial cells by centrifugation; 3) mixing the bacterial cells with skim milk and freeze-drying the mixture.
[0026] Furthermore, the inoculation amount of the Lactobacillus salivarius PC114 is 1.25×10 7 ~1×10 8 CFU / mL.
[0027] Furthermore, the liquid culture medium may be MRS liquid culture medium.
[0028] Preferably, in step 1), the fermentation process is static culture, the fermentation temperature is 25-45° C., and the fermentation time is 12 to 36 hours.
[0029] More preferably, the temperature can be selected from 25°C, 27°C, 29°C, 31°C, 33°C, 35°C, 37°C, 39°C, 41°C, 43°C, 45°C, etc., preferably 37°C; the time can be selected from 12h, 14h, 16h, 18h, 20h, 22h, 24h, 26h, 28h, 30h, 32h, 34h, 36h, etc., preferably 24h.
[0030] Preferably, in step 2), the centrifugal speed is 4000-12000 g, and the time is 8-12 min.
[0031] More preferably, the rotation speed can be selected from 4000g, 4500g, 5000g, 5500g, 6000g, 6500g, 7000g, 7500g, 8000g, 8500g, 9000g, 9500g, 10000g, 10500g, 11000g, 11500g, 12000g, etc., preferably 10000g; the time can be selected from 8min, 8.5min, 9min, 9.5min, 10min, 10.5min, 11min, 11.5min, 12min, etc., preferably 10min.
[0032] Preferably, the mass percentage of skim milk powder in the skim milk is 6-12%, for example, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, 10%, 10.5%, 11%, 11.5%, 12%, etc., preferably 10%.
[0033] In a preferred embodiment, the viable cell count of the Lactobacillus salivarius PC114 strain is not less than 1×10 9 CFU / g, for example 1×10 9 CFU / g, 2×10 9 CFU / g, 3×10 9 CFU / g, 4×10 9 CFU / g, 5×10 9 CFU / g, 6×10 9 CFU / g, 7×10 9 CFU / g, 8×10 9 CFU / g, 9×10 9 CFU / g, 1×10 10 CFU / g, 2×10 10 CFU / g, 5×10 10 CFU / g, 8×10 10 CFU / g, 1×10 11 CFU / g, 5×10 11 CFU / g, 1×10 12 CFU / g, 1×10 13 CFU / g, etc. Other specific point values within the above numerical range can be selected and will not be described here one by one.
[0034] The deposit information of the strains in this application is as follows:
[0035] Strain name: Lactobacillus salivarius;
[0036] The accession number is: CGMCC NO.23518;
[0037] Deposit date: September 29, 2021;
[0038] Name of depository institution: General Microbiology Center of China Culture Collection Administration of Microorganisms;
[0039] Abbreviation of depository unit: CGMCC;
[0040] Address of the depository: No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.
[0041] The experimental methods in the following examples, unless otherwise specified, are all conventional methods. The experimental materials used in the following examples, unless otherwise specified, were purchased from conventional biochemical reagent companies. The quantitative experiments in the following examples were all repeated three times, and the results were averaged.
[0042] Example 1
[0043] Preparation of seeds (fermentation bacteria)
[0044] A single colony of the strain was picked with an inoculating loop and placed in 10 mL of MRS liquid medium (purchased from Merck Co., Germany). The colony was evenly dispersed in the liquid medium using a vortex shaker and incubated anaerobically at 37°C for 48 h. The culture was taken out and inoculated into MRS liquid medium at a 2% (v / v) inoculum size. After anaerobically incubated at 37°C for 24 h, the culture was centrifuged at 15,000 rpm for 10 min, and the supernatant was discarded. The bacteria were washed twice with sterile distilled water and then suspended in sterile distilled water of the original culture volume to obtain seeds for fermentation. The bacterial concentration of the seed liquid was 2.5×10 9 CFU / mL.
[0045] Example 2
[0046] Preparation of microbial agents
[0047] Preparation method of skim milk: mix 10% by mass of skim milk powder with distilled water, fully dissolve it, sterilize it at 125°C for 5 minutes, and cool it to room temperature to obtain skim milk of the required concentration.
[0048] Preparation method of microbial agent:
[0049] (1) The fermentation strain seed liquid of Lactobacillus salivarius PC114 (CGMCC No. 23518) was aseptically inoculated into MRS liquid medium at an inoculum rate of 2% (v / v, the volume percentage of the seed liquid to the fermentation liquid, the same below) and incubated at 37°C for 24 h to obtain a fermentation liquid;
[0050] (2) The fermentation broth obtained in step (1) was centrifuged at 10,000 g for 10 min, the supernatant was discarded, and the precipitate (bacteria) was washed twice with sterile distilled water;
[0051] (3) Resuspend the bacterial cells obtained in step (2) with skim milk (the mass percentage of the vegetation in the material preparation is 10%) of the original culture volume.
[0052] Example 3
[0053] Promoting effect of Lactobacillus salivarius PC114 on steroid hormone synthesis in aged mice receiving fecal microbiota transplantation
[0054] Animal experiment: 16 GF-grade adult C57BL / 6 mice (Jiangsu Jicui Yaokang Biotechnology Co., Ltd.) were purchased and randomly divided into 4 groups of 4 mice each. The feces of 5 6-week-old and 20-month-old male C57BL / 6 mice were mixed evenly to prepare fecal microbiota transplantation solutions for young and old mice. The feces were gavaged once a day for 3 times, including 2 groups gavaged with young mouse fecal microbiota transplantation solutions and 2 groups gavaged with old mouse fecal microbiota transplantation solutions. One group was gavaged with PC114 (viable bacteria count of 1×10 9 CFU / mL) 0.2mL for two weeks, and another group was gavaged with skim milk as a control.
[0055] The experimental animal information and gavage method are shown in Table 1.
[0056] Table 1
[0057]
[0058] (1) Impact on metabolic pathways
[0059] The KEGG metabolic pathway association analysis was performed on the fecal metabolome samples of the aged mice fecal microbiota transplantation group and the aged mice fecal microbiota transplantation + PC114 intervention group using the MetaboAnalyst platform (version 6.0). The main regulated metabolic pathways in the non-targeted metabolome results were as follows: Figure 1 As shown in the data, the main metabolic pathway regulated during PC114 intervention was the biosynthesis pathway of steroid hormones. The experimental results showed that Lactobacillus salivarius PC114 could significantly regulate the steroid hormone biosynthesis process of germ-free mice transplanted with fecal bacteria from old mice.
[0060] (2) Effects on the content of specific steroid hormones and their derivatives
[0061] By analyzing the significantly regulated steroid hormones and their derivatives in non-targeted metabolomics, such as Figure 2As shown in the study, pregnenolone levels in the young mice group receiving fecal microbiota transplantation were significantly higher than those in the old mice group. After 14 days of intervention with Lactobacillus salivarius PC114, pregnenolone levels in the old mice group were significantly increased, approaching or even exceeding those in the young mice group. Pregnenolone is a precursor to androgen synthesis and, as a source of testosterone, increases testosterone production.
[0062] In addition, if Figure 3 As shown in the results, after 14 days of intervention with Lactobacillus salivarius PC114, the content of 11β-hydroxyandrostenedione in the fecal microbiota transplantation group of elderly mice was significantly increased. This substance is a steroid specific to the adrenal gland with androgenic activity. Under the necessary enzymatic mechanism, it will promote the formation of new androgen C11-oxoandrogen.
[0063] Figure 4 It shows that 14 days of PC114 intervention can significantly increase the content of 19-oxoandrost-4-ene-3,17-dione, bringing it close to the level of the young mouse transplant group. According to the annotation information of the HMDB database, this substance belongs to androgens and their derivatives, which are beneficial to the development of male characteristics, such as promoting the growth of hair and body hair.
[0064] In summary, Lactobacillus salivarius PC114 can significantly regulate the steroid hormone biosynthesis pathway in aged mice, increasing the levels of pregnenolone, 11β-hydroxyandrostenedione, and 19-oxoandrost-4-ene-3,17-dione, making them closer to those of young mice.
[0065] The applicant declares that the present invention illustrates the application of a Lactobacillus salivarius of the present invention in preparing a product for enhancing male hormones in the elderly through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvements to the present invention, equivalent replacements of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the scope of protection and disclosure of the present invention.
[0066] While the basic concepts have been described above, it will be apparent to those skilled in the art that the detailed disclosure is merely illustrative and does not limit this specification. Although not explicitly stated herein, various modifications, improvements, and revisions to this specification may be made by those skilled in the art. Such modifications, improvements, and revisions are suggested in this specification and remain within the spirit and scope of the exemplary embodiments of this specification.
[0067] This specification also uses specific terms to describe the embodiments of this specification. For example, "one embodiment," "an embodiment," and / or "some embodiments" refer to a feature, structure, or characteristic associated with at least one embodiment of this specification. Therefore, it should be emphasized and noted that references to "one embodiment," "an embodiment," or "an alternative embodiment" two or more times in different locations in this specification do not necessarily refer to the same embodiment. Furthermore, certain features, structures, or characteristics of one or more embodiments of this specification may be appropriately combined.
[0068] Finally, it should be understood that the embodiments described in this specification are intended only to illustrate the principles of the embodiments of this specification. Other variations may also fall within the scope of this specification. Therefore, by way of example and not limitation, alternative configurations of the embodiments of this specification may be considered consistent with the teachings of this specification. Accordingly, the embodiments of this specification are not limited to the embodiments explicitly described and illustrated in this specification.
Claims
1. An application of Lactobacillus salivarius in the preparation of a product for enhancing male hormone secretion in the elderly, characterized in that: The Lactobacillus salivarius is the Lactobacillus salivarius PC114 strain with a preservation number of CGMCC No.23518.
2. The use according to claim 1, characterized in that Products that enhance male hormone secretion in the elderly have one or more of the following functions: 1) increasing the level of androgens and their derivatives, preferably increasing the level of pregnenolone, 11β-hydroxyandrostenedione and 19-oxoandrost-4-ene-3,17-dione; 2) Regulate steroid biosynthesis signaling pathways.
3. The use according to claim 1, characterized in that The product includes one or more of health products, foods or medicines; preferably, the medicine also includes a pharmaceutically acceptable carrier or excipient.
4. The use according to claim 3, characterized in that The dosage form of the product includes one or more of injection, suspension, powder, tablet or granule.
5. The use according to claim 1, characterized in that The viable count of Lactobacillus salivarius PC114 in the product is not less than 1×10 9 CFU / g.
6. The use according to any one of claims 1 to 5, characterized in that: The product contains Lactobacillus salivarius PC114 bacterial agent.
7. The use according to claim 6, characterized in that The preparation method of the Lactobacillus salivarius PC114 bacterial agent comprises the following steps: 1) inoculating Lactobacillus salivarius strain PC114 into a liquid culture medium for fermentation and culture; 2) separating and collecting the bacterial cells cultured in step 1), and collecting the bacterial cells by centrifugation; 3) Mixing the bacterial cells obtained in step 2) with skim milk and freeze-drying the mixture.
8. The use according to claim 7, characterized in that The inoculation amount of the Lactobacillus salivarius strain PC114 was 1.25×10 7 ~1×10 8 CFU / mL.
9. The use according to claim 7, characterized in that The fermentation process is static culture, the fermentation temperature is 25-45° C., and the fermentation time is 12 to 36 hours.
10. The use according to claim 7, characterized in that The mass percentage of skim milk powder in the skim milk is 6-12%; And / or, the centrifugal speed is 4000-12000g, and the time is 8-12min.