Application of L-beta-galactoglucan in anti-vaginitis aspect

Abstract

The invention provides a novel application of L-beta-galactoglucan, and particularly provides an application of the L-beta-galactoglucan in the aspect of resisting vaginitis.

Description

TECHNICAL FIELD

[0001] The present application relates to a new use of L-beta-galactosan in health care and medicine, and in particular to the application of L-beta-galactosan in the treatment of vaginitis. BACKGROUND

[0002] Vaginitis is an inflammation of the mucosa and submucosal connective tissue of the vagina, and is one of the most common diseases in gynecology. Its core feature is that the balance of the normal microecological environment in the vagina is broken. Under normal circumstances, there are a variety of microorganisms in the vagina, mainly lactobacilli, which are in harmony with each other and maintain a healthy acidic environment in the vagina (pH value is usually 3.8-4.4). When this balance is disrupted for various reasons, pathogenic bacteria or opportunistic pathogens (such as Candida) will overproliferate, or exogenous pathogens will invade, thereby triggering an inflammatory response and a series of symptoms.

[0003] Vaginitis is mainly classified according to the pathogenic cause, and the most common types include: (1) Bacterial vaginosis, which is not an "inflammation" caused by a single bacterium, but a dysbiosis of the vaginal flora. The number of lactobacilli is greatly reduced, while other anaerobes (such as Gardnerella, mobiluncus) overproliferate.

[0004] (2) Vulvovaginal candidiasis, an infection caused by the overgrowth of fungi (mainly Candida albicans).

[0005] (3) Trichomonas vaginitis, a sexually transmitted infection caused by the protozoan Trichomonas vaginalis.

[0006] (4) Atrophic vaginitis, a non-infectious inflammation. It is more common in postmenopausal or low estrogen women, due to the decline of ovarian function and the decrease of estrogen levels, leading to vaginal wall atrophy, thinning of the mucosa, and decreased local resistance.

[0007] (5) Other types: including vaginitis caused by specific pathogens such as gonococci, mycoplasma, and chlamydia, as well as non-infectious vulvovaginal inflammation caused by allergies or irritants (such as soap, condoms, and washing liquids).

[0008] WO2025002125A1 discloses an L-beta-galactosan produced by Agrobacterium and its preparation method and application. The repeating unit structure of the galactosan is as follows: .

[0009] The activity of the L-beta-galactosan in the treatment of vaginitis has not been disclosed. SUMMARY

[0010] The present application aims to study the activity of L-beta-galactosan in resisting vaginitis.

[0011] Specifically, one aspect of the present application is to provide the use of L-beta-galactosan or crude polysaccharide containing L-beta-galactosan in the preparation of a product resisting Candida.

[0012] In some embodiments of the present application, the Candida is selected from Candida albicans.

[0013] In some embodiments of the present application, the product is a product treating the vaginal mucosa damage caused by Candida.

[0014] In some embodiments of the present application, the product can improve the abnormal vaginal pH caused by Candida.

[0015] In some embodiments of the present application, the product is a product improving the microecological balance in the vagina.

[0016] In some embodiments of the present application, the product inhibits the appearance of Proteobacteria in the vaginal environment, and induces the appearance of Gemmatimonadetes, Chloroflexi or / and Acidobacteria.

[0017] In some embodiments of the present application, the product is a product treating the inflammation of the vagina caused by Candida.

[0018] In some embodiments of the present application, the product is a product reducing the inflammatory cell infiltration in the inflammation of the vagina, and inhibiting the abnormal increase of inflammatory factors.

[0019] In some embodiments of the present application, the inflammatory factors are selected from IL-1β or / and IL-6.

[0020] In some embodiments of the present application, the preparation method of the L-beta-galactosan comprises the following steps: (1) inoculating Agrobacterium FN01 in a culture solution to perform fermentation to obtain a fermentation liquor; (2) alcohol precipitation of the fermentation liquor, drying to obtain a crude polysaccharide product, dissolving the crude polysaccharide product, adding alkali and filter aid to perform filtration, alcohol precipitation and drying of the filtrate to obtain a pure polysaccharide product; or performing enzymolysis of the fermentation liquor with glucanase, and then performing filtration, ultrafiltration concentration and spray drying to obtain a pure polysaccharide product.

[0021] In some embodiments of the present application, the preparation method of the crude polysaccharide containing L-beta-galactosan comprises the following steps: (1) inoculating Agrobacterium FN01 in a culture solution to perform fermentation to obtain a fermentation liquor; (2) alcohol precipitation of the fermentation liquor, drying to obtain a crude polysaccharide product.

[0022] In another aspect of the present application, there is provided use of L-beta-galactosan or crude polysaccharide containing L-beta-galactosan in the preparation of a medicament for treating vaginal candidiasis.

[0023] In another aspect of the present application, there is provided use of L-beta-galactosan or crude polysaccharide containing L-beta-galactosan in the preparation of a product for improving the microecological balance in the vagina.

[0024] The present application has found that, in a vaginitis model, administration of L-beta-galactosan can effectively reduce the content of inflammatory factors IL-6 and IL-1beta, eliminate Candida albicans in the vagina, and improve vaginal mucosal damage, and can regulate the diversity of vaginal microbial flora and the abundance of pathogenic bacteria, and maintain the microecological balance. BRIEF DESCRIPTION OF DRAWINGS

[0025] Figure 1 Vulva observation pictures.

[0026] Figure 2 Colony counting results A: bacterial concentration; B: colony clearance rate; C: colony counting plate picture; ns: compared with the blank group; ns: compared with the model group; n=6.

[0027] Figure 3 ELISA detection results of rat vaginal lavage fluid A: IL-1beta; B: IL-6; ns: compared with the blank group, p * <0.05; compared with the model group, p # <0.05, p ## <0.01. n=6.

[0028] Figure 4 Pathological score results of rat vagina A: inflammatory cell infiltration score; B: epithelial damage score; ns: compared with the blank group, p * <0.05; compared with the model group, p ## <0.01. n=6.

[0029] Figure 5 Alpha diversity index of rat vaginal microorganisms, wherein Blank: blank group; Model: model group; High: high concentration of Ru Rukuo group; ns: compared with the blank group, p * <0.05, compared with the model group, ns.

[0030] Figure 6 Alpha diversity index of vaginal microorganisms in rats, wherein Blank: blank group; Model: model group; High: high concentration group of Fururukoka.

[0031] Figure 7 Analysis of rat vaginal flora at the door level, wherein yellow is the main affected bacteria door in the model group, and blue is the newly introduced bacteria door in the high concentration group of Fururukoka. Blank: blank group; Model: model group; High: high concentration group of Fururukoka.

[0032] Figure 8 Analysis of rat vaginal flora at the genus level DETAILED DESCRIPTION

[0033] The L-beta-galactosan used in the present application is provided by Chengdu Saideke Biological Technology Co., Ltd. (trade name: Fururukoka) and is prepared according to Example 1 in WO2025002125A1. The repeating unit of L-beta-galactosan is shown in structural formula I:

[0034] Formula I It can also be as follows: .

[0035] The auxiliary materials of the present application are the general term of all additional materials except the main components, and the auxiliary materials should have the following properties: (1) no toxic effect on human body, few side effects; (2) stable chemical properties, not easy to be affected by temperature, pH, storage time, etc.; (3) no compatibility contraindication with the main drug, and no influence on the efficacy and quality inspection of the main drug; (4) no mutual action with the packaging materials. The auxiliary materials in the present application include but are not limited to fillers (diluents), lubricants (glidants or anti-adhesion agents), dispersants, wetting agents, binders, adjusting agents, solubilizers, antioxidants, bacteriostatic agents, emulsifiers, disintegrants, etc. The binders include syrup, gum arabic, gelatin, sorbitol, tragacanth gum, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethyl cellulose, ethyl cellulose or hydroxypropyl methyl cellulose, etc.), gelatin syrup, sugar syrup, starch paste or polyvinylpyrrolidone, etc.; the fillers include lactose, sugar powder, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salt (such as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; the lubricants include micro-powder silica gel, magnesium stearate, talc, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol, etc.; the disintegrants include starch and its derivatives (such as sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, etc.), polyvinylpyrrolidone or microcrystalline cellulose, etc.; the wetting agents include sodium dodecyl sulfate, water or alcohol, etc.; the antioxidants include sodium sulfite, sodium bisulfite, sodium metabisulfite, dibutyl phenyl acid, etc.; the bacteriostatic agents include 0.5% phenol, 0.3% methyl phenol, 0.5% trichlorobutanol, etc.; the adjusting agents include hydrochloric acid, citric acid, potassium (sodium) hydroxide, sodium citrate and buffer (including sodium dihydrogen phosphate and disodium hydrogen phosphate), etc.; the emulsifiers include polysorbate-80, sorbic acid sorbitan, pluronic F-68, lecithin, soybean phospholipid, etc.; the solubilizers include tween-80, bile, glycerol, etc.

[0036] Oral solid products, which include capsules, tablets, pills, powders and granules in their use forms. In these solid products, the active compound is mixed with at least one conventional inert excipient (or carrier), such as sodium citrate or dicalcium phosphate, or with the following ingredients: (a) fillers or extenders, e.g., starches, lactose, sucrose, glucose, mannitol, and silicic acid; (b) binders, e.g., hydroxymethylcellulose, alginates, gelatin, polyvinylpyrrolidone, sucrose, and acacia; (c) humectants, e.g., glycerol; (d) disintegrating agents, e.g., agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain complex silicates, and sodium carbonate; (e) solution retarders, e.g., paraffin; (f) absorption accelerators, e.g., quaternary ammonium compounds; (g) wetting agents, e.g., cetyl alcohol and glycerol monostearate; (h) adsorbents, e.g., kaolin and bentonite; and (i) lubricants, e.g., talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, or mixtures thereof. In the capsules, tablets and pills, a buffering agent can also be included.

[0037] Vaginal administration forms include, but are not limited to, suppositories, vaginal tablets / effervescent tablets, soft capsules, creams, gels, lotions, etc. Among them, suppositories can melt at body temperature to release drugs, have smooth effect and are widely used. Vaginal tablets / effervescent tablets disintegrate and release in the moist environment of the vagina, sometimes producing bubbles to promote distribution. Soft capsules have a gelatin shell, containing oily or paste-like drugs, which cover the mucosa after melting. Creams and gels are semisolid and can be injected into the vagina or applied to the vulva. Lotions are liquids used for cleaning, wet compressing or sitz bath, and have the functions of auxiliary cleaning and relieving discomfort.

[0038] Common excipients in external administration forms include, but are not limited to: (1) penetration enhancers, such as amines, oleic acid, propylene glycol, laurocapram, menthol, isopropyl myristate; (2) pH adjusters, such as lactic acid, citric acid, sodium hydroxide, boric acid, sodium phosphate / dibasic sodium phosphate, triethanolamine; (3) effervescent agents, sodium bicarbonate + organic acid (such as citric acid, tartaric acid, fumaric acid); (4) preservatives, parabens (nipagin), sodium benzoate, potassium sorbate, benzalkonium chloride, chlorobutanol; (5) thickening / suspending agents, such as magnesium aluminum silicate, xanthan gum, hydroxypropyl methyl cellulose, carbomer, sodium alginate; (6) lubricants, such as magnesium stearate, silicon dioxide, microcrystalline cellulose, talc, polyethylene glycol (PEG); (7) antioxidants, such as tocopherol; etc.

[0039] In some embodiments of the present invention, the content of L-β-galactoglucan, or crude polysaccharide containing L-β-galactoglucan, in the product is 0.1% or more, calculated as L-β-galactoglucan as shown in Formula I, including but not limited to 0.1%, 0.2%, 0.3%, 0.4%, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1%, 2%, 3%, 4%, 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, etc.

[0040] In some embodiments of the present invention, the content of L-β-galactoglucan represented by Formula I in the product is above 0.01%.

[0041] In some embodiments of the present invention, the content of L-β-galactoglucan represented by Formula I in the product is 0.1% or more.

[0042] The present invention also provides a method for treating vaginitis, which involves applying an effective amount of L-β-galactoglucan or crude polysaccharide containing L-β-galactoglucan into the vagina.

[0043] The present invention also provides a method for adjuvant treatment of vaginitis, which involves applying an effective amount of L-β-galactoglucan or crude polysaccharide containing L-β-galactoglucan to the vulva.

[0044] The effective dose, calculated based on L-β-galactoglucan, refers to the dose required to produce a clear and expected therapeutic effect when using L-β-galactoglucan or crude polysaccharides containing L-β-galactoglucan. It is not a fixed value, but rather a statistical range related to the probability of response in a population.

[0045] Treatment refers to interventions performed on individuals with diseases or health problems in order to cure the disease, control the condition, relieve symptoms, restore function, prevent the disease or complications, and / or improve the quality of life. Example 1

[0046] 1. Animal grouping Thirty-six SPF-grade female SD rats were acclimatized for 7 days and then randomly divided into a control group (n=6) and a model group (n=30). After successful modeling, the model groups were randomly divided into a model group, a low-concentration fumarole group, a medium-concentration fumarole group, a high-concentration fumarole group, and a clotrimazole group, with n=6.

[0047] 2. Preparation of bacterial culture 2.1 Strain Culture Candida albicans was activated on YPD medium for 24 h, and 1 mL of the bacterial suspension was added to RPMI-1640 medium and cultured for 24 h until it reached the mycelial stage.

[0048] 2.2 Preparation of bacterial culture Centrifuge the bacterial culture, collect the precipitate, resuspend it in PBS, determine the concentration using the McFarland turbidimetric method, and adjust the bacterial concentration to 1.5 × 10⁻⁶ using sterile PBS. 8 CFU / mL, for later use.

[0049] 3. Model preparation and drug intervention Animals in the model group were subcutaneously injected with 0.1 mL of estradiol benzoate injection solution starting 6 days before inoculation, once every 2 days for a total of 3 injections, with day 1 being the day of inoculation. 1.5 × 10 8 CFU / mL Candida albicans bacterial suspension (hyphae stage) was used to instill 50 μL of the bacterial suspension into the vagina of rats in the model group after rinsing the vagina three times with physiological saline. The suspension was left in place for 1-2 minutes to prevent leakage. The control group received an equal volume of sterile physiological saline instilled into the vagina. On day 4, if vulvar redness and swelling and increased secretions were observed in the rats, vaginal secretions were collected for microscopic examination. The presence of fungal spores or hyphae under the microscope confirmed successful model establishment.

[0050] After successful model establishment, rats in the low-concentration, medium-concentration, and high-concentration groups of fumarole were administered fumarole vaginally at doses of 2.10 mg / kg, 4.20 mg / kg, and 8.40 mg / kg, respectively, with a dosage of 0.32 mL / kg. The positive control group was administered clotrimazole vaginally at a dose of 15.75 mg / kg, while the blank control and model groups were given an equal volume of purified water. Rats in each group were inverted for 5 minutes after administration to ensure the drug did not leak from the vaginal opening. The day of the first administration was designated as day 0, and administration continued from day 0 to day 6, once daily for 7 consecutive days.

[0051] 4 Evaluation Indicators 4.1 Observation of general clinical characteristics During the experiment, the general clinical symptoms (diet, mental state, gait, hair, etc.) of the experimental animals were observed once a day, and the vagina was photographed on days 1, 3, 5, and 7 after administration.

[0052] 4.2 Weight monitoring Weigh yourself once a week and record your weight.

[0053] 4.3 Colony Count On day 7 after drug administration, the vagina was irrigated five times with 500 μL of sterile saline. 50 μL of the irrigation fluid was taken, diluted 10-fold, and 50 μL of the diluted solution was spread on YPD agar medium. This process was repeated three times, and the mixture was incubated at 37°C for 48 h. Colony counts were performed, and the colony clearance rate was calculated using the following formula: Colony clearance rate (%) = (mean bacterial concentration in the model group - mean bacterial concentration in the drug group) / mean bacterial concentration in the model group × 100 4.4 ELISA Detection On day 7 after administration, 150 μL of lavage fluid was collected, and the levels of inflammatory factors IL-1β and IL-6 in the vaginal lavage fluid were measured according to the IL-1β and IL-6 kit instructions.

[0054] 4.5 Pathological Examination On day 7 after drug administration, a portion of vaginal tissue was collected, fixed with 10% neutral formaldehyde, dehydrated, cleared, embedded in paraffin, sectioned, dewaxed with xylene, dehydrated with graded ethanol, stained with hematoxylin, differentiated with hydrochloric acid and alcohol, stained with eosin, dehydrated with graded ethanol, cleared with a clearing agent, and mounted with neutral resin. Images of the sections were acquired using a Pannoramic 250 digital section scanner, and the tissue lesions were observed under a light microscope. The severity of the lesions was evaluated using a scoring system. The scoring criteria are shown in Table 1.

[0055] Table 1 Scoring Criteria

[0056] 4.6 Microbial community sequencing On day 7 after drug administration, vaginal secretions from the vaginal opening of the blank group, model group, and high-concentration Furuike group were collected using sterile cotton swabs. Total DNA was extracted and quantified. The target fragment was amplified by PCR, and the amplified products were purified and recovered by magnetic beads. The PCR amplified products were quantitatively analyzed by fluorescence. Sequencing libraries were prepared using the TruSeq Nano DNA LT Library Prep Kit and sequenced using a MiSeq sequencer. The observed species, Chao1, Shannon, Simpson, and Pielou's sevenness indices of vaginal flora were compared among the groups to assess the richness and diversity of the flora. The relative abundance at the phylum and genus levels among the groups was compared, and principal coordinate analysis was performed.

[0057] 5. Statistical Analysis All experimental data were entered and analyzed using statistical software (e.g., Excel, SPSS, etc.). All measurement data were expressed as mean ± standard deviation. This indicates that homogeneity of variance was tested among the dose groups and the control group for the measurement data. When the variances are homogeneous ( p>(0.05), the Dunnett's & LSD method in one-way ANOVA was used to compare differences between groups. When variances are unequal ( P For values ​​≤0.05, the Mann-Whitney U test (MW test) within the Kruskal-Wallis H rank-sum test (KW method) was used to compare differences between groups. For ordinal data, nonparametric tests were performed separately for different genders. p <0.05, p <0.01, p <0.001 indicates that the difference is statistically significant.

[0058] 6. Experimental Results 6.1 Observation results of general clinical characteristics like Figure 1 As shown, during the experiment, the control group rats were in normal condition. After inoculation with Candida albicans, the model group showed vulvar redness and swelling and increased secretions. On days 1, 3, and 5 of administration, the redness and swelling of the low, medium, and high concentration groups of Furuike and the clotrimazole group gradually subsided, and the secretions decreased or disappeared. Among them, the medium and high concentration groups of Furuike showed more significant improvement in the symptoms of redness and swelling and increased secretions than the low concentration group of Furuike. On day 7 of administration, there were no obvious symptoms of redness and swelling and increased secretions in the vagina of the low, medium, and high concentration groups of Furuike and the clotrimazole group, while the model group still had symptoms of redness and swelling and increased secretions.

[0059] The above results suggest that low, medium, and high concentrations of phenobarbital can all improve the symptoms of redness, swelling, and increased secretions in rats with vaginal mucosal injury.

[0060] 6.2 Weight monitoring results During the experiment, the weight of rats in the blank group increased steadily, while the weight of rats in the model group, low-, medium-, and high-concentration ferrous sulfate groups, and clotrimazole group decreased to some extent after modeling, with significantly lower weights on days 4, 8, and 11 compared to the blank group. p <0.05); After administration, the body weight of the low, medium, and high concentration groups of Furuike and the clotrimazole group showed an increasing trend, with the body weight on day 11 (day 7 of administration) being higher than that of the model group, with a significant difference in the high concentration group of Furuike. p <0.05). The above results suggest that low, medium, and high concentrations of phenobarbital can all reverse the trend of weight loss in rats with vaginal mucosal injury.

[0061] 6.3 Colony Count Results The concentration of Candida albicans in the vaginal irrigation fluid of the model group rats was significantly higher than that in the control group. Compared with the model group, the concentrations of Candida albicans in the low, medium, and high concentrations of fluocinolone acetonide and the clotrimazole group were significantly reduced. Among them, the medium and high concentrations of fluocinolone acetonide and the clotrimazole group showed the best antibacterial effects, with colony clearance rates exceeding 70%. See details below. Figure 2 .

[0062] The results above suggest that low, medium, and high concentrations of fluocinolone can clear vaginal Candida albicans in rats with vaginal mucosal injury, with medium and high concentrations showing better clearance effects.

[0063] 6.4 ELISA test results ELISA results showed that, compared with the blank group, the levels of IL-1β and IL-6 in the model group showed an increasing trend, with IL-1β showing a significant difference. p <0.05); Compared with the model group, the IL-1β and IL-6 levels in the low, medium, and high concentration groups of fumarole and the clotrimazole group showed a decreasing trend, with the IL-1β levels in the low, high concentration groups of fumarole and the clotrimazole group being significantly lower ( p <0.05 or p <0.01), the IL-6 content in the FUruike medium concentration group was significantly reduced ( p <0.05). See details below. Figure 3 .

[0064] The results above suggest that low, medium, and high concentrations of phenobarbital can effectively reduce the levels of IL-1β and IL-6 in rats with vaginal mucosal injury and improve inflammation.

[0065] 6.5 Pathological examination results Pathological examination results showed that the vaginal mucosal epithelium of the control group rats was normal. In the other groups, the most prominent lesion in the vagina was squamous epithelial mucosal metaplasia. A large number of squamous epithelium were replaced by mucosal cells, resulting in vaginal epithelial mucosification. Some mucosal cells degenerated and died, appearing as vacuoles. The clotrimazole group was the most severe, followed by the model group and the low-concentration fluocinolone group. The high-concentration fluocinolone group was the next most severe, and the medium-concentration fluocinolone group had the mildest degeneration. Some groups also had a certain amount of epithelial cell necrosis and shedding. At the same time, inflammatory cell infiltration, mainly polymorphonuclear leukocytes, could be observed in the mucosal epithelium, with some spreading into the lamina propria. The inflammatory response in the control group was slightly milder, while the inflammatory response in the model group and the low-concentration fluocinolone group was slightly more severe. The inflammatory response in the medium-concentration fluocinolone group, the high-concentration fluocinolone group, and the clotrimazole group was lower than that in the model group and the low-concentration fluocinolone group.

[0066] Pathological scoring results showed that, compared with the control group, the inflammatory cell infiltration score and epithelial damage score of the model group were significantly increased. p <0.05); Compared with the model group, there were no significant changes in the inflammatory cell infiltration score and epithelial damage score in the low-concentration group of fumarole, while the inflammatory cell infiltration score was somewhat reduced in the high-concentration group of fumarole and the clotrimazole group. The epithelial damage score was significantly reduced in the medium- and high-concentration groups of fumarole, with a significant difference in the medium-concentration group of fumarole. p<0.01), the epithelial damage score in the clotrimazole group was elevated to some extent, which may be related to drug irritation. See details below. Figure 4 .

[0067] The above results suggest that medium and high concentrations of fluocinolone acetonide can effectively reduce vaginal tissue damage in a rat model of vaginal mucosal injury.

[0068] 6.6 Results of microbial community sequencing 6.6.1 Alpha Diversity Analysis like Figure 6 As shown, all Alpha diversity indices (Observed species, Chao1, Shannon, Simpson, Pielou's evenness) in the model group were significantly increased, with Observed species and Chao1 showing significant differences. p <0.05). In contrast, all indices in the high-concentration group of Furuike showed a decreasing trend, with the Chao1 and Observed species indices showing the most significant decreasing trend.

[0069] The above results suggest that the vaginal flora of rats with vaginal mucosal injury model is in a typical state of "pathological high diversity". High concentration of fluocinolone can effectively inhibit the excessive growth of pathogenic bacteria and opportunistic pathogens, and promote the return of the microbial community from a chaotic state to a healthy low diversity state, suggesting that high concentration of fluocinolone has the potential to regulate the vaginal microecological balance.

[0070] 6.6.2 Principal Coordinate Analysis (PCoA) PCoA analysis results showed a significant difference in distribution between the blank group and the model group. p <0.01 indicates that the vaginal microbial community structure in the model group has changed significantly, deviating from the normal state; the distribution location of the high-concentration Furuike group is significantly far away from that in the model group ( p <0.05), with no significant difference in distance from the control group, indicating that the community structure of the high-concentration furuticosa group was significantly restored. See details below. Figure 6 .

[0071] The above results suggest that Furuike can regulate the vaginal microbial community structure in a rat model of vaginal mucosal injury.

[0072] 6.6.3 Phyllostachyal and Genus-Level Analysis Phylum-level analysis showed that the blank group mainly consisted of Proteobacteria, Actinobacteria, and Firmicutes; the model group showed a significant increase in the relative abundance of Proteobacteria, Firmicutes, and Bacteroidetes, and a significant decrease in the relative abundance of Actinobacteria; the high-concentration group of Furuike significantly reduced the relative abundance of Proteobacteria, and also induced Gemmatimonadetes, Chloroflexi, and Acidobacteria. See details below. Figure 7 .

[0073] Genus-level analysis showed that, compared with the control group, the relative abundance of *Streptococcus* was significantly reduced in the model group, while the relative abundance of *Citrobacter* and *Aerococcus* was significantly increased. High-concentration phenylephrine in the model group improved this phenomenon. See details below. Figure 8 .

[0074] The above results suggest that the rat model of vaginal mucosal injury exhibits a significant imbalance in vaginal microecology. Specifically, at the phylum level, the relative abundance of Proteobacteria (a phylum closely related to inflammation, rich in Gram-negative bacteria that release lipopolysaccharide LPS, a key substance that strongly triggers the release of pro-inflammatory factors such as IL-1β and IL-6) was significantly increased, which is highly consistent with the results of ELISA detection of soaring levels of pro-inflammatory factors in vaginal douches. At the same time, the relative abundance of Firmicutes (containing beneficial bacteria such as Lactobacillus and opportunistic pathogens such as Staphylococcus) and Bacteroidetes (opportunistic pathogens) also increased significantly, while the relative abundance of Actinobacteria (a beneficial flora that helps stabilize a healthy vagina, whose decrease in abundance is often accompanied by a decrease in lactic acid production and an increase in pH) decreased significantly. This change in the microbiota structure directly led to an increase in vaginal pH and a loss of the microecological barrier function.

[0075] At the genus level, the relative abundance of *Streptococcus* (normally colonizing bacteria) decreased, while the relative abundance of *Citrobacter* (belonging to the phylum Proteobacteria, a typical opportunistic pathogen that can induce a strong inflammatory response) and *Gastrodia* (opportunistic pathogens) increased. This change is highly consistent with the pathological mechanisms in vaginal mucosal injury models, namely, disruption of the microecological barrier dominated by *Lactobacillus*, overgrowth of opportunistic pathogens, and increased susceptibility to *Candida albicans*.

[0076] The high-concentration group of Furuike effectively regulated the aforementioned bacterial flora imbalance. Its mechanism of action is manifested on two levels: First, it directly and significantly reduced the relative abundance of the core harmful phylum—Proteobacteria—reducing pro-inflammatory stimulation from the source and providing a direct bacterial explanation for the decrease in IL-1β and IL-6 levels in vaginal irrigation fluid. Second, it induced the appearance of Bacillus, Chlorella, and Acidobacteria, which were not detected in the blank and model groups. These newly introduced phyla, especially Acidobacteria, which prefer acidic environments, help maintain and consolidate the normal acidic environment of the vagina after treatment by filling ecological niches and leveraging their acid-producing / acid-resistant characteristics. They can also compete with harmful bacteria for nutrients and attachment sites, thereby indirectly optimizing the vaginal microecological structure. At the genus level, it can reverse the decrease in Streptococcus and the increase in Citrobacter and Gastrodia. These results collectively demonstrate that high concentrations of FURUKEN have the potential to repair vaginal microecological imbalance, inhibit the spread of opportunistic pathogens, and ultimately improve the stability of the vaginal flora structure in a multi-target, multi-level manner in a model of vaginal mucosal damage.

[0077] in conclusion: Under the conditions of this experiment, low (2.10 mg / kg), medium (4.20 mg / kg), and high (8.40 mg / kg) concentrations of phenobarbital (Fluorococcus) could, to some extent, reduce the levels of inflammatory factors IL-6 and IL-1β in rats with vaginal mucosal injury, effectively clear Candida albicans from the vagina of the rat model of vaginal mucosal injury, and improve vaginal mucosal injury. The mechanism may be related to the abnormal increase in vaginal microbiota diversity and pathogen abundance in the rat model of vaginal mucosal injury, suggesting that Fluorococcus has a role in regulating vaginal flora structure and maintaining microecological balance.

Claims

1. Use of L-β-galactoglucan or crude polysaccharides containing L-β-galactoglucan in the preparation of products against Candida spp.

2. The use according to claim 1, characterized in that, The Candida species mentioned are selected from Candida albicans.

3. The use according to claim 1 or 2, characterized in that, The product is for treating vaginal mucosal damage caused by Candida albicans.

4. The use according to claim 1 or 2, characterized in that, The product can improve vaginal pH abnormalities caused by Candida albicans.

5. The use according to claim 1 or 2, characterized in that, The product described is designed to improve the balance of the vaginal microecology.

6. The use according to claim 1, characterized in that, The product inhibits Proteobacteria in the vaginal environment and induces the appearance of Bacillus, Curvularia, and / or Acidobacteria.

7. The use according to claim 1, characterized in that, The product is for treating vaginal inflammation caused by Candida.

8. The use according to claim 1, characterized in that, The product described herein is designed to reduce inflammatory cell infiltration and inhibit abnormal increases in inflammatory factors during vaginal inflammation.

9. The use according to claim 8, characterized in that, The inflammatory factors are selected from IL-1β and / or IL-6.

10. Use of L-β-galactoglucan or crude polysaccharides containing L-β-galactoglucan in the preparation of medicines for treating vaginal candidiasis and / or bacterial vaginosis.

11. Use of L-β-galactoglucan or crude polysaccharides containing L-β-galactoglucan in the preparation of products that improve the vaginal microecological balance.

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