A probiotic active antibacterial tablet for preventing and treating female reproductive system infection and preparation method thereof

Abstract

The present invention provides a probiotic active antibacterial tablet for preventing and treating female reproductive system infection and a preparation method thereof. The present invention is a probiotic active antibacterial tablet for preventing and treating female reproductive system infection, comprising the following raw materials in parts by weight: 1.8-2.5 parts of Lactobacillus reuteri, 1.0-1.2 parts of Lactobacillus rhamnosus, 1.2-1.7 parts of Lactobacillus crispatus, 0.5-0.8 parts of Lactobacillus gasseri, 1.0-1.5 parts of stachyose, 0.8-1.2 parts of inulin, and 2.5-3 parts of plant extract, wherein the plant extract is prepared from the following raw materials: Bletilla striata, Chinese tallow tree leaf, Gentiana scabra, and grapefruit seed. The probiotic active antibacterial tablet prepared by the present invention has a significant inhibitory effect on IGFBP7 protein, can effectively prevent and treat female reproductive system infection, and has good tolerance and safety.

Description

Technical Field

[0001] The present invention relates to the field of antibacterial preparations, and in particular to a probiotic active antibacterial tablet for preventing and treating female reproductive system infections and a preparation method thereof. Background Art

[0002] Female reproductive tract infections are common, including bacterial vaginosis, candidal vaginosis, and Trichomonas vaginosis. These infections can cause symptoms such as vaginitis, vaginal itching, and increased vaginal discharge, severely impacting women's quality of life and health. Conventional treatments include antibiotics and antifungal drugs, but long-term use can be harmful due to the high risk of developing drug resistance and dependence.

[0003] As a safe and effective treatment, probiotics can regulate the reproductive system's microbiome and maintain physiological balance, making them a promising new option for treating female reproductive tract infections. However, the current market offers a wide variety of probiotic products with varying quality and clinical efficacy. Therefore, developing a safe and effective probiotic active antibacterial tablet tailored to the specific characteristics of female reproductive tract infections is crucial for addressing the challenges of traditional treatments. Summary of the Invention

[0004] In view of this, the present invention provides a probiotic active antibacterial tablet for preventing and treating female reproductive system infections and a preparation method thereof.

[0005] The technical solution of the present invention is achieved as follows:

[0006] A probiotic active antibacterial tablet for preventing and treating female reproductive system infections comprises the following raw materials in parts by weight: 1.8-2.5 parts of Lactobacillus reuteri, 1.0-1.2 parts of Lactobacillus rhamnosus, 1.2-1.7 parts of Lactobacillus crispatus, 0.5-0.8 parts of Lactobacillus gasseri, 1.0-1.5 parts of stachyose, 0.8-1.2 parts of inulin, and 2.5-3 parts of a plant extract, wherein the plant extract is prepared from the following raw materials: bletilla striata, Chinese tallow tree leaves, gentian, and grapefruit seeds.

[0007] Preferably, the plant extract is prepared from the following raw materials in parts by weight: 10-20 parts of Bletilla striata, 3-5 parts of Chinese tallow tree leaves, 8-12 parts of Gentiana scabra, and 20-25 parts of grapefruit seeds.

[0008] Preferably, the method for preparing the plant extract comprises the following steps:

[0009] (1) According to the weight ratio, the Chinese medicinal materials of Bletilla striata, Chinese tallow tree leaf, and Gentiana scabra were respectively taken, crushed into powder, extracted with subcritical water at 138-140°C for 20-30 minutes, with a material-liquid ratio of g / ml of 1:15-20, and the Chinese medicinal extract was collected;

[0010] (2) Grapefruit seeds were added with 90%-95% v / v ethanol for reflux extraction according to weight ratio, with a solid-liquid ratio of g / ml of 1:10-15, an extraction temperature of 80-90°C, and an extraction time of 2-3h. The grapefruit seed extract was collected and concentrated under reduced pressure to obtain a grapefruit seed extract;

[0011] (3) The Chinese medicine extract and the grapefruit seed extract are mixed evenly, and freeze-dried to obtain the target plant extract.

[0012] Preferably, the probiotic active antibacterial tablet comprises the following raw materials in parts by weight: 2 parts of Lactobacillus reuteri, 1 part of Lactobacillus rhamnosus, 1.5 parts of Lactobacillus crispatus, 0.5 parts of Lactobacillus gasseri, 1.2 parts of stachyose, 1 part of inulin, and 1.5 parts of plant extract.

[0013] Preferably, the viable count of Lactobacillus reuteri is 3 to 4 billion CFU / g, the viable count of Lactobacillus rhamnosus is 8 to 9 billion CFU / g, the viable count of Lactobacillus crispatus is 10 to 20 billion CFU / g, and the viable count of Lactobacillus gasseri is 5 to 6 billion CFU / g.

[0014] The preparation method of the probiotic active antibacterial tablet of the present invention comprises the following steps:

[0015] Step 1: Evenly mix Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus crispatus, and Lactobacillus gasseri to obtain mixed probiotics;

[0016] Step 2: Mix stachyose, inulin and purified water at a material-liquid ratio of 1:8-10 g / ml, and stir evenly to obtain a mixed solution;

[0017] Step 3: Mix the mixed probiotics and plant extracts, add the mixed solution to the mixture in step 2, and vacuum homogenize;

[0018] Step 4: vacuum-dry the homogenized material obtained in step 3, and press the tablets to obtain probiotic active antibacterial tablets.

[0019] Preferably, in step 2, the stirring speed is 800-1200 rpm, the stirring temperature is 40-50° C., and the stirring time is 10-20 min.

[0020] Preferably, in step 3, the vacuum homogenization conditions are: vacuuming to -0.05 to -0.03 MPa, homogenization temperature of 35-40° C., rotation speed of 1000-2000 r / min, and homogenization time of 8-12 min.

[0021] Compared with the prior art, the present invention has the following beneficial effects:

[0022] (1) The probiotic active antibacterial tablets prepared by the present invention have a significant inhibitory effect on IGFBP7 protein. Among them, the present invention adopts a certain proportion of Lactobacillus gasseri and Lactobacillus reuteri, Lactobacillus rhamnosus, and Lactobacillus crispatus to effectively enhance the inhibitory effect on IGFBP7 protein of Candida albicans vaginitis. The present invention adopts a certain proportion of grapefruit seeds, white ginseng, Chinese tallow leaves, and gentian, and combines composite bacteria to further improve the inhibitory effect on IGFBP7 protein of Candida albicans vaginitis. The present invention adopts sucrose with inulin to better exert the efficacy of composite bacteria and plant extracts, and effectively promotes the improvement of the inhibitory level of IGFBP7 protein of Candida albicans vaginitis.

[0023] (2) The probiotic active antibacterial tablets prepared by the present invention have good antibacterial effects on Candida albicans and Staphylococcus aureus.

[0024] (3) The probiotic active antibacterial tablets of the present invention can stimulate the immune system cell response in the body, strengthen the reproductive system, maintain an acidic pH value in the reproductive tract, effectively inhibit the growth of harmful bacteria, and reduce the risk of infection. The probiotic active antibacterial tablets of the present invention can effectively prevent and treat female reproductive system infections and have good tolerability and safety. DETAILED DESCRIPTION

[0025] In order to better understand the technical content of the present invention, specific examples are provided below to further illustrate the present invention.

[0026] Unless otherwise specified, the experimental methods used in the examples of the present invention are all conventional methods.

[0027] Unless otherwise specified, the materials, reagents, etc. used in the examples of the present invention can be obtained from commercial sources.

[0028] The Lactobacillus gasseri, Lactobacillus reuteri, Lactobacillus rhamnosus and Lactobacillus crispatus used in the examples of the present invention are all commercially available bacterial agents (bacterial powders).

[0029] Preparation Example 1

[0030] The plant extract is prepared from the following raw materials in parts by weight: 15 parts of Bletilla striata, 4 parts of Chinese tallow tree leaves, 10 parts of Gentiana scabra, and 23 parts of grapefruit seeds. The preparation method of the plant extract comprises the following steps:

[0031] (1) According to the weight ratio, the Chinese medicinal materials of Bletilla striata, Chinese tallow tree leaf, and Gentiana scabra were respectively taken and ground into powder, and extracted with subcritical water at 138-140°C for 25 min, with a material-liquid ratio of g / ml of 1:18, and the Chinese medicinal extract was collected;

[0032] (2) Grapefruit seeds were added with 95% v / v ethanol for reflux extraction according to weight ratio, with a solid-liquid ratio of g / ml of 1:12, an extraction temperature of 85°C ± 5°C, and an extraction time of 2.5 h. The grapefruit seed extract was collected and concentrated under reduced pressure to obtain a grapefruit seed extract;

[0033] (3) The Chinese medicine extract and the grapefruit seed extract are mixed evenly, and freeze-dried to obtain the target plant extract.

[0034] Example 1 - Preparation of probiotic active antibacterial tablets

[0035] (1) Formula

[0036] The raw materials for the probiotic active antibacterial tablets are calculated by weight as follows: 2.0 parts of Lactobacillus reuteri, 1.0 part of Lactobacillus rhamnosus, 1.5 parts of Lactobacillus crispatus, 0.8 part of Lactobacillus gasseri, 1.2 parts of stachyose, 1.0 part of inulin, and 2.5 parts of the plant extract of Preparation Example 1. The viable cell count of Lactobacillus reuteri is approximately 3.5 billion CFU / g, the viable cell count of Lactobacillus rhamnosus is 8.5 billion CFU / g, the viable cell count of Lactobacillus crispatus is 15 billion CFU / g, and the viable cell count of Lactobacillus gasseri is 5.5 billion CFU / g.

[0037] (2) Process

[0038] Step 1: Evenly mix Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus crispatus, and Lactobacillus gasseri to obtain mixed probiotics;

[0039] Step 2: Mix stachyose, inulin and purified water at a material-liquid ratio of 1:9 g / ml, stir at a stirring speed of 1000 rpm, a stirring temperature of 45°C ± 5°C, and a stirring time of 15 minutes to obtain a mixed solution;

[0040] Step 3: Mix the mixed probiotics and plant extracts, add the mixed solution to the mixed solution in step 2, and vacuum homogenize to -0.04 MPa. The homogenization temperature is 38°C ± 2°C, the speed is 1500 r / min, and the homogenization time is 10 min.

[0041] Step 4: vacuum-dry the homogenized material obtained in step 3, and press the tablets to obtain probiotic active antibacterial tablets.

[0042] Example 2-Preparation of probiotic active antibacterial tablets

[0043] (1) Formula

[0044] The raw materials for the probiotic active antibacterial tablets are calculated by weight as follows: 2.5 parts of Lactobacillus reuteri, 1.0 part of Lactobacillus rhamnosus, 1.3 parts of Lactobacillus crispatus, 0.5 part of Lactobacillus gasseri, 1.0 part of stachyose, 1.0 part of inulin, and 2.9 parts of the plant extract of Preparation Example 1. The viable cell count of Lactobacillus reuteri is approximately 3.5 billion CFU / g, the viable cell count of Lactobacillus rhamnosus is 8.5 billion CFU / g, the viable cell count of Lactobacillus crispatus is 15 billion CFU / g, and the viable cell count of Lactobacillus gasseri is 5.5 billion CFU / g.

[0045] (2) Process

[0046] Step 1: Evenly mix Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus crispatus, and Lactobacillus gasseri to obtain mixed probiotics;

[0047] Step 2: Mix stachyose, inulin and purified water at a material-liquid ratio of 1:9 g / ml, stir at a stirring speed of 1000 rpm, a stirring temperature of 45°C ± 5°C, and a stirring time of 15 minutes to obtain a mixed solution;

[0048] Step 3: Mix the mixed probiotics and plant extracts, add the mixed solution to the mixed solution in step 2, and vacuum homogenize to -0.04 MPa. The homogenization temperature is 38°C ± 2°C, the speed is 1500 r / min, and the homogenization time is 10 min.

[0049] Step 4: vacuum-dry the homogenized material obtained in step 3, and press the tablets to obtain probiotic active antibacterial tablets.

[0050] Example 3 - Preparation of probiotic active antibacterial tablets

[0051] (1) Formula

[0052] The raw materials for the probiotic active antibacterial tablets are calculated by weight as follows: 1.8 parts of Lactobacillus reuteri, 1.2 parts of Lactobacillus rhamnosus, 1.7 parts of Lactobacillus crispatus, 0.6 parts of Lactobacillus gasseri, 1.5 parts of stachyose, 1.0 part of inulin, and 2.2 parts of the plant extract of Preparation Example 1. The viable cell count of Lactobacillus reuteri is approximately 3.5 billion CFU / g, the viable cell count of Lactobacillus rhamnosus is 8.5 billion CFU / g, the viable cell count of Lactobacillus crispatus is 15 billion CFU / g, and the viable cell count of Lactobacillus gasseri is 5.5 billion CFU / g.

[0053] (2) Process

[0054] Step 1: Evenly mix Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus crispatus, and Lactobacillus gasseri to obtain a mixed probiotic;

[0055] Step 2: Mix stachyose, inulin and purified water at a material-liquid ratio of 1:9 g / ml, stir at a stirring speed of 1000 rpm, a stirring temperature of 45°C ± 5°C, and a stirring time of 15 minutes to obtain a mixed solution;

[0056] Step 3: Mix the mixed probiotics and plant extracts, add the mixed solution to the mixed solution in step 2, and vacuum homogenize to -0.04 MPa. The homogenization temperature is 38°C ± 2°C, the speed is 1500 r / min, and the homogenization time is 10 min.

[0057] Step 4: vacuum-dry the homogenized material obtained in step 3, and press the tablets to obtain probiotic active antibacterial tablets.

[0058] Comparative Example 1

[0059] The main difference from Example 1 is that Lactobacillus gasseri is not contained, but the total amount of bacterial powder added remains unchanged.

[0060] (1) Formula

[0061] The raw materials of the probiotic active antibacterial tablets are calculated by weight: 2.0 parts of Lactobacillus reuteri, 1.5 parts of Lactobacillus rhamnosus, 1.8 parts of Lactobacillus crispatus, 1.2 parts of stachyose, 1.0 part of inulin, and 2.5 parts of plant extracts.

[0062] (2) Process

[0063] The same as Example 1.

[0064] Comparative Example 2

[0065] The main difference from Example 1 is that the amount of each bacterial powder added is different from that in Example 1.

[0066] (1) Formula

[0067] The raw materials of the probiotic active antibacterial tablets are calculated by weight: 1.4 parts of Lactobacillus reuteri, 1.3 parts of Lactobacillus rhamnosus, 1.3 parts of Lactobacillus crispatus, 1.3 parts of Lactobacillus gasseri, 1.2 parts of stachyose, 1.0 part of inulin, and 2.5 parts of plant extracts.

[0068] (2) Process

[0069] The same as Example 1.

[0070] Comparative Example 3

[0071] The main difference from Example 1 is that the plant extract does not contain Bletilla striata and grapefruit seeds, but the amount of plant extract added remains unchanged.

[0072] (1) Formula

[0073] The raw materials of the probiotic active antibacterial tablets are calculated by weight: 2.0 parts of Lactobacillus reuteri, 1.0 part of Lactobacillus rhamnosus, 1.5 parts of Lactobacillus crispatus, 0.8 part of Lactobacillus gasseri, 1.2 parts of stachyose, 1.0 part of inulin, and 2.5 parts of plant extracts.

[0074] (2) Process

[0075] The same as Example 1.

[0076] Comparative Example 4

[0077] The main difference from Example 1 is that stachyose is replaced by sucrose.

[0078] (1) Formula

[0079] The raw materials of the probiotic active antibacterial tablets are calculated by weight: 2.0 parts of Lactobacillus reuteri, 1.0 part of Lactobacillus rhamnosus, 1.5 parts of Lactobacillus crispatus, 0.8 part of Lactobacillus gasseri, 1.2 parts of sucrose, 1.0 part of inulin, and 2.5 parts of plant extract.

[0080] (2) Process

[0081] The same as Example 1.

[0082] Test Example 1 Antibacterial Test

[0083] Experimental groups 1-7: corresponding to the use of the probiotic active antibacterial tablets of Examples 1-3 and Comparative Examples 1-4, respectively.

[0084] Set up blank control group (CK): normal saline.

[0085] Test method: Staphylococcus aureus was inoculated on a slant Sabouraud medium, and Candida albicans was inoculated on a slant Sabouraud medium, and they were cultured at 37 ° C for 48 hours, and the bacterial solution concentration was cultured to 300 million CFU / mL, and the bacterial solution was prepared for use. 100 μL of bacterial solution was inoculated onto the corresponding agar plate, and then 1 mg of the probiotic active antibacterial tablets (crushed to powder) of Examples 1-3 and Comparative Examples 1-4 were added and evenly distributed. An equal amount of normal saline was added to the blank control group. Place in a 37 ° C constant temperature incubator, culture for 24 hours, and observe the results. The diameter of the inhibition zone was measured by the cross-cross method, repeated 3 times, and the average value was taken.

[0086] Table 1 Antibacterial test results

[0087]

[0088] The results in Table 1 show that the probiotic active antibacterial tablets prepared in Examples 1-3 have good antibacterial effects on Candida albicans and Staphylococcus aureus.

[0089] Test Example 2 - Mouse Test

[0090] 100 female SPF Kunming mice were selected, 10 of which were randomly selected as blank control group, and 90 mice were used for modeling. The modeling method was to subcutaneously inject estradiol benzoate injection 0.1 ml / d to put the mice in pseudoestrus state, and 2.5×10 6 cfu / mL bacterial suspension 20 μL (i.e., the inoculum size of Candida albicans spores is 5×10 4 CFU) were instilled into the vaginas of mice. Model identification was performed on mice 2-5 days after inoculation with Candida albicans spores: 5 mice were randomly selected twice for vulvar observation, microscopic examination, and sacrificed for vaginal pathological tissue sections. The results showed vulvovaginal swelling and purulent secretions. Microscopic examination revealed numerous spores and hyphae. Vaginal pathological tissue sections showed congestion and redness, confirming successful modeling. After successful modeling, the mice were randomly divided into the following groups: a model control group and study groups 1-7 (using the probiotic active antibacterial tablets of Examples 1-3 and Comparative Examples 1-4, respectively), with 10 mice in each group.

[0091] Dosing: Study groups 1-7 were administered intravaginally with a 1 mg powdered probiotic tablet once daily for 7 consecutive days. The blank and model control groups were not administered intravaginally but received an equal volume of normal saline. The day after 7 days of dosing, the mice were sacrificed by cervical dislocation, and vaginal mucosa samples were collected for Western blot analysis of IGFBP7 protein levels in each group. The effects of the probiotic tablet on IGFBP7 expression in mice with Candida albicans vaginitis were evaluated.

[0092] Table 2 Effect of probiotic active antibacterial tablets on IGFBP7 protein content

[0093]

[0094] The results in Table 1 show that the probiotic active antibacterial tablets prepared in Examples 1-3 have a significant inhibitory effect on IGFBP7 protein.

[0095] The inhibitory effect of Comparative Example 1-2 was significantly reduced compared with Example 1. The present invention uses a certain ratio of Lactobacillus gasseri to Lactobacillus reuteri, Lactobacillus rhamnosus and Lactobacillus crispatus to effectively enhance the inhibitory effect of IGFBP7 protein on Candida albicans vaginitis.

[0096] The plant extract of Comparative Example 3 does not contain Bletilla striata and grapefruit seeds, and its inhibitory effect is also significantly reduced. The present invention uses a certain proportion of grapefruit seeds, Bletilla striata, Chinese tallow leaves, and Gentiana scabra to further improve the inhibitory effect of IGFBP7 protein on Candida albicans vaginitis.

[0097] In the comparative example 4, stachyose was not used, but sucrose was added, and the inhibitory effect was also reduced. The present invention uses sucrose with inulin to better exert the efficacy of the composite bacteria and plant extracts, effectively promoting the improvement of the inhibition level of IGFBP7 protein in Candida albicans vaginitis.

[0098] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent replacements, improvements, etc. made within the principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A probiotic active antibacterial tablet for preventing and treating female reproductive system infections, characterized in that: The invention is composed of the following raw materials in parts by weight: 1.8-2.5 parts of Lactobacillus reuteri, 1.0-1.2 parts of Lactobacillus rhamnosus, 1.2-1.7 parts of Lactobacillus crispatus, 0.5-0.8 parts of Lactobacillus gasseri, 1.0-1.5 parts of stachyose, 0.8-1.2 parts of inulin, and 2.5-3 parts of plant extracts. The plant extracts are prepared from the following raw materials in parts by weight: 10-20 parts of Bletilla striata, 3-5 parts of Sapium sebiferum leaves, 8-12 parts of Gentiana scabra, and 20-25 parts of grapefruit seeds.

2. The probiotic active antibacterial tablet according to claim 1, characterized in that The preparation method of the plant extract comprises the following steps: (1) Take the Chinese medicinal materials of Bletilla striata, Sapium sebiferum leaves and Gentiana scabra respectively according to the weight ratio, grind them into powder, extract them with subcritical water at 138-140℃ for 20-30min, and the material-liquid ratio g / ml is 1:15-20, and collect the Chinese medicinal extract; (2) Grapefruit seeds were added with 90%-95% v / v ethanol for reflux extraction according to weight ratio, with a solid-liquid ratio of g / ml of 1:10-15, an extraction temperature of 80-90°C, and an extraction time of 2-3h. The grapefruit seed extract was collected and concentrated under reduced pressure to obtain grapefruit seed extract; (3) The Chinese herbal medicine extract and grapefruit seed extract are mixed evenly, and freeze-dried to obtain the target plant extract.

3. The probiotic active antibacterial tablet according to claim 1 or 2, characterized in that The invention is composed of the following raw materials in parts by weight: 2 parts of Lactobacillus reuteri, 1 part of Lactobacillus rhamnosus, 1.5 parts of Lactobacillus crispatus, 0.5 part of Lactobacillus gasseri, 1.2 parts of stachyose, 1 part of inulin and 1.5 parts of plant extract.

4. The probiotic active antibacterial tablet according to claim 1, characterized in that The viable bacteria count of the Lactobacillus reuteri is 3 to 4 billion CFU / g, the viable bacteria count of the Lactobacillus rhamnosus is 8 to 9 billion CFU / g, the viable bacteria count of the Lactobacillus crispatus is 10 to 20 billion CFU / g, and the viable bacteria count of the Lactobacillus gasseri is 5 to 6 billion CFU / g.

5. The method for preparing the probiotic active antibacterial tablet according to any one of claims 1 to 4, characterized in that: The following steps are involved: Step 1: Evenly mix Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus crispatus, and Lactobacillus gasseri to obtain mixed probiotics; Step 2: Mix stachyose, inulin and purified water at a material-liquid ratio of 1:8-10 g / ml, and stir evenly to obtain a mixed solution; Step 3: Mix the mixed probiotics and plant extracts, add the mixed solution to the mixture in step 2, and vacuum homogenize; Step 4: vacuum-dry the homogenized material obtained in step 3, and press the tablets to obtain probiotic active antibacterial tablets.

6. The method for preparing the probiotic active antibacterial tablet according to claim 5, characterized in that: In step 2, the stirring speed is 800-1200 rpm, the stirring temperature is 40-50°C, and the stirring time is 10-20 min.

7. The method for preparing the probiotic active antibacterial tablet according to claim 5, characterized in that: In step 3, the vacuum homogenization conditions are: vacuuming to -0.05 to -0.03 MPa, homogenization temperature of 35-40° C., rotation speed of 1000-2000 r / min, and homogenization time of 8-12 min.

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