Vaginal mucosa repairing composition containing rose component and application

Abstract

According to the vaginal mucosa repairing composition containing the rose component and the application, the vaginal mucosa repairing composition achieves multiple effects of inhibiting pathogens, relieving inflammation and repairing mucosa through the synergistic effect of the LGG fermentation product and the rose component. According to the in-vitro verification method based on the vaginal mucosa epithelial cell 3D model, an in-vivo multi-layer mucosa structure and a complex microenvironment are accurately simulated, the barrier function, inflammation regulation and control and cell proliferation capacity of the vaginal mucosa repair composition are accurately evaluated, and the mucosa barrier function and multi-cell interaction are reflected.

Description

Technical Field

[0001] The invention belongs to the technical field of female reproductive health care, and relates to a vaginal mucosa repairing composition containing a rose component and an application thereof. Background Art

[0002] Vaginal itching and mucosal damage are common gynecological problems in women, often caused by factors such as bacterial or fungal vaginitis, hormonal changes, poor hygiene, or chemical irritation. Clinical manifestations include vaginal mucosal congestion, edema, and unbearable itching. In severe cases, it can lead to mucosal damage, pain, and even affect quality of life.

[0003] Currently, the main treatments for vaginal itching and mucosal damage include antibiotics, hormonal drugs, and traditional Chinese medicine. Antibiotics and hormonal drugs can quickly control infection, but long-term use can easily lead to drug resistance and secondary imbalance of the microbiome. Traditional Chinese medicine has anti-inflammatory and antibacterial potential, but its mechanism of action is complex and the efficacy of a single ingredient is limited. In recent years, probiotics have garnered attention for their ability to regulate microecological balance. However, when used, probiotics only play a role in regulating the vaginal flora and do not participate in the repair of mucosal damage. It is impossible to achieve the "itch relief + mucosal repair" problem using probiotics alone. Summary of the Invention

[0004] The purpose of the present invention is to provide a vaginal mucosa repairing composition containing rose ingredients and its application, so as to solve the problem that existing probiotics cannot achieve "itch relief + mucosa repair".

[0005] To achieve the above object, the present invention adopts the following technical solutions: The present application provides a vaginal mucosa repair composition containing a rose component, wherein the composition comprises 50-80% of a Lactobacillus rhamnosus fermentation product and 20-50% of a rose component in terms of mass percentage.

[0006] Lactobacillus rhamnosus fermentation product (LGG-FP) is produced by anaerobic fermentation of Lactobacillus rhamnosus (LGG). Specifically, Lactobacillus rhamnosus LGG is added to MRS medium (DeMan, Rogosa, and Sharpe medium) and anaerobically fermented at 37°C for 18-24 hours to produce a fermentation broth. After centrifugation to remove bacterial cells, the broth is filtered through a 0.22μm pore size filter membrane to obtain a supernatant. This supernatant is then concentrated or freeze-dried to obtain the Lactobacillus rhamnosus fermentation product. When the Lactobacillus rhamnosus fermentation product is a concentrated solution, its solids content is 10-15%.

[0007] The active ingredients contained in the fermentation products of Lactobacillus rhamnosus mainly include exopolysaccharides (EPS) with a molecular weight of 10-500kDa, short-chain fatty acids (SCFAs) such as lactic acid, acetic acid, propionic acid, gamma-aminobutyric acid (GABA) and bacteriocins such as LGG-1.

[0008] Rose ingredients include one or more of rose water extract, rose essential oil or rose polysaccharides. Its main active ingredients are polyphenols such as gallic acid and quercetin, flavonoids such as apigenin and luteolin, volatile oils such as citronellol and phenylethanol, which have anti-inflammatory, antioxidant and epithelial cell proliferation-promoting effects.

[0009] The rose component in this application is preferably a rose water extract, wherein the rose water extract has a solid content of 5-10%. The preparation method of the rose water extract comprises adding cleaned rose petals to distilled water, heating and refluxing at 80°C for 2 hours, and filtering and concentrating the extract to a solid content of 5-10%.

[0010] In this application, the vaginal mucosa repair composition synergistically exerts the dual effects of "itch relief and repair" through the fermentation product of Lactobacillus rhamnosus and rose ingredients. The synergistic mechanism is: (1) Inhibit the adhesion and proliferation of pathogens: The extracellular polysaccharides in LGG-FP can form a biofilm on the surface of the vaginal mucosa, competitively inhibiting the adhesion sites of pathogens such as Escherichia coli and Candida albicans, thereby inhibiting the reproduction and growth of pathogens such as Escherichia coli and Candida albicans; rose polyphenols can downregulate the expression of pathogen virulence genes such as ompA and cdx, and enhance the microecological regulation ability of LGG-FP.

[0011] (2) Relieve inflammatory response: SCFAs in LGG-FP can reduce the local pH value in the vagina and maintain it at 4.5-5.5, inhibiting the release of inflammatory factors such as IL-6 and TNF-α; rose flavonoids such as luteolin can inhibit the NF-κB signaling pathway and reduce the secretion of itching mediators such as histamine and 5-HT.

[0012] (3) Promote mucosal repair: GABA in LGG-FP can activate the PI3K / Akt pathway of vaginal epithelial cells and promote cell proliferation; rose polysaccharides can upregulate the expression of tight junction proteins such as ZO-1 and Occludin and mucus secretion proteins such as MUC1, thereby repairing the damaged mucosal barrier.

[0013] More preferably, the composition comprises 60-70% of Lactobacillus rhamnosus fermentation product and 30-40% of rose component in terms of mass percentage.

[0014] The vaginal mucosal repair composition provided herein can be used to prepare a medicament for treating vaginal mucosal repair. The medicament can be formulated as a topical gel, suppository, or lotion. When formulating the vaginal mucosal repair composition into a medicament, the ratio of carriers such as carbomer, glycerin, and deionized water needs to be adjusted according to the formulation to ultimately achieve a dosage form suitable for vaginal administration.

[0015] In addition, the vaginal mucosal repair composition provided in the present application can also be used to evaluate in vitro efficacy based on a 3D model of vaginal mucosal epithelial cells.

[0016] Specifically, the method for evaluating in vitro efficacy based on the vaginal mucosal epithelial cell 3D model includes: S01: Human vaginal epithelial cells were seeded onto Matrigel or acellular vaginal matrix scaffolds, added to DMEM / F12 culture medium containing 10% fetal bovine serum, and cultured in a 37°C, 5% CO2 incubator for 7-10 days to form a 3D model of vaginal mucosal epithelial cells. The Matrigel contains basement membrane components such as laminin and type IV collagen. S02: setting up a negative control group, a positive control group, an experimental group, a fermentation product group alone, and a rose component group alone, wherein the negative control group includes a culture medium, the positive control group includes 1 μg / mL lipopolysaccharide, the experimental group includes the above-mentioned vaginal mucosa repair composition, the fermentation product group alone includes a Lactobacillus rhamnosus fermentation product with a solid content of 10%, and the rose component group alone includes a rose component with a solid content of 10%; S03: adding the negative control group, positive control group, experimental group, fermentation product single group, and rose component single group to the vaginal mucosal epithelial cell 3D model, respectively, to detect transepithelial electrical resistance, inflammatory factor secretion, and histological structure; S04: When the transepithelial electrical resistance of the experimental group is ≥ that of the negative control group, and the inflammatory factor secretion content is ≤ that of the negative control group, the vaginal mucosa repair composition has mucosa repair and antipruritic effects.

[0017] The present invention has the following beneficial effects: (1) The vaginal mucosal repair composition in this application achieves the multiple effects of "inhibiting pathogens, relieving inflammation, and repairing mucosa" through the synergistic effect of LGG fermentation products and rose ingredients.

[0018] (2) This application is based on an in vitro verification method of a 3D model of vaginal mucosal epithelial cells, which accurately simulates the multi-layered mucosal structure and complex microenvironment in vivo, accurately evaluates the barrier function, inflammation regulation and cell proliferation ability of the vaginal mucosal repair composition, and reflects the mucosal barrier function and multicellular interactions. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is a graph showing the effect of the vaginal mucosal repair composition prepared in Example 1 of the present application on the TEER value of the LPS injury model, *p<0.05; Figure 2 This is a diagram showing the inhibitory effect of the vaginal mucosa repair composition prepared in Example 1 of the present application on IL-6 secretion; Figure 3 This is a diagram showing the inhibitory effect of the vaginal mucosa repair composition prepared in Example 1 of the present application on TNF-α secretion; Figure 4 This is the HE staining image of the vaginal mucosa repair composition prepared in Example 1 of the present application on the LPS injury model. DETAILED DESCRIPTION

[0020] The technical solution of the present invention is further explained and illustrated by means of specific embodiments below.

[0021] Example 1 The embodiment of the present application provides a vaginal mucosa repair composition containing a rose component, which comprises 60% of a Lactobacillus rhamnosus fermentation product and 40% of a rose flower water extract in terms of mass percentage.

[0022] The present embodiment also uses the above-mentioned vaginal mucosal repair composition containing rose ingredients to prepare a topical gel matrix. The preparation method of the topical gel matrix includes: S101: Lactobacillus rhamnosus LGG is added to MRS medium at a 10% inoculum and anaerobically fermented at 37°C for 18-24 hours to obtain a fermentation broth. The fermentation broth is centrifuged at 8000 g for 10 minutes to remove bacterial cells, and then filtered through a 0.22 μm pore size filter membrane to obtain a supernatant. The supernatant is concentrated to 1 / 3 of its original volume to obtain a Lactobacillus rhamnosus fermentation product with a solids content of 10%.

[0023] S102: The washed rose petals were mixed with deionized water in a mass-to-volume ratio of 1:10, and the mixture was heated under reflux at 80° C. for 2 h. The extract was filtered and concentrated to a rose water extract having a solid content of 8%.

[0024] S103: After uniformly mixing the fermentation product of Lactobacillus rhamnosus and the water extract of rose flower in a mass ratio of 7:3, 0.5% Carbomer 940 was added as a thickener, the mixture was made up to 100% with deionized water, and the pH was adjusted to 5.0 to obtain a topical gel matrix.

[0025] Example 2 The embodiment of the present application provides a vaginal mucosa repair composition containing a rose component, which comprises 70% of a Lactobacillus rhamnosus fermentation product and 30% of a rose flower water extract in percentage by mass.

[0026] Example 3 The embodiment of the present application provides a vaginal mucosa repair composition containing a rose component, which comprises 50% of a Lactobacillus rhamnosus fermentation product and 50% of rose essential oil by mass percentage.

[0027] Example 4 The embodiment of the present application provides a vaginal mucosa repair composition containing rose ingredients, which comprises 80% of Lactobacillus rhamnosus fermentation product and 20% of rose polysaccharide in terms of mass percentage.

[0028] Example 5 The present invention provides a method for evaluating in vitro efficacy based on a 3D model of vaginal mucosal epithelial cells, the method comprising: S501: Human vaginal epithelial cells VK2 / E6E7 were cultured at a rate of 5×10 4 Cells were seeded at a rate of 10 cells / well into Matrigel-coated Transwell chambers. DMEM / F12 culture medium supplemented with 10% fetal bovine serum was added to the chambers and cultured in a 37°C, 5% CO2 incubator for 7 days to form a 3D model of vaginal mucosal epithelial cells. HE staining revealed clear stratification of the basal cell spinous layer and the surface cell layer, indicating successful 3D model establishment.

[0029] S502: Set up a negative control group, a positive control group, an experimental group, a fermentation product single-component group, and a rose component single-component group, wherein the negative control group includes culture medium without the addition of other substances; the positive control group includes culture medium + 1 μg / mL lipopolysaccharide; the experimental group includes culture medium + vaginal mucosa repair compositions with different final concentrations, wherein the final concentrations of the vaginal mucosa repair compositions are 1%, 5%, and 10%, respectively; the fermentation product single-component group includes culture medium + Lactobacillus rhamnosus fermentation product with a solid content of 10%; the rose component single-component group includes culture medium + rose component with a solid content of 10%.

[0030] S503: Add negative control group, positive control group, experimental group, fermentation product group and rose component group to the 3D model of vaginal mucosal epithelial cells, and detect transepithelial resistance, inflammatory factor secretion, and histological structure to obtain the attached Figure 1-4 Among them, the detection indicators of transepithelial electrical resistance (TEER), inflammatory factor secretion, itch mediator content and histological structure are shown in Table 1.

[0031] Table 1: Detection indicators and detection methods S04: When the transepithelial electrical resistance of the experimental group is ≥ that of the negative control group, and the inflammatory factor secretion content is ≤ that of the negative control group, the vaginal mucosa repair composition has mucosa repair and antipruritic effects.

[0032] By the attached Figure 1-3 It can be seen that compared with the negative control group, the TEER of the experimental group was significantly increased, and the secretion of inflammatory factors IL-6 and TNF-α was significantly reduced, which shows that the vaginal mucosa repair composition prepared in Example 1 has the effect of relieving itching of the mucosa. Figure 4 It can be seen that compared with the negative control, the vaginal mucosal epithelial cells of the 10% composition are more compact and the tissue structure is more complete, indicating that the composition has a mucosal repair effect. The above content shows that the vaginal mucosa repair composition prepared in the examples of the present application can significantly improve the barrier function of the vaginal mucosa, inhibit vaginal inflammatory response and promote cell proliferation, confirming that it has the dual effects of "relieving vaginal itching and repairing vaginal mucosa".

[0033] The above description is merely a preferred embodiment of the present application and is not intended to limit the present application. Various modifications and variations are possible for those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present application shall be included within the scope of protection of the present application.

Claims

1. A vaginal mucosa repairing composition containing rose ingredients, characterized in that: The composition comprises 50-80% of Lactobacillus rhamnosus fermentation product and 20-50% of rose component in terms of mass percentage.

2. The vaginal mucosa repairing composition containing rose ingredients according to claim 1, characterized in that: According to mass percentage, the composition comprises 60-70% of Lactobacillus rhamnosus fermentation product and 30-40% of rose component.

3. The vaginal mucosa repairing composition containing rose ingredients according to claim 1, characterized in that: The rose component includes one or more of rose water extract, rose essential oil or rose polysaccharide.

4. The vaginal mucosa repairing composition containing rose ingredients according to claim 3, characterized in that: The solid content of the rose water extract is 5-10%.

5. The vaginal mucosa repairing composition containing rose ingredients according to claim 1, characterized in that: The Lactobacillus rhamnosus is anaerobically fermented at 37° C. for 18-24 hours, and a Lactobacillus rhamnosus fermentation product is obtained after centrifugation, filtration, concentration or freeze-drying.

6. The vaginal mucosa repair composition according to any one of claims 1 to 5 is used for preparing a medicament for treating vaginal mucosa repair.

7. The use according to claim 6, wherein the drug is in the form of a topical gel, suppository or lotion.

8. The vaginal mucosa repair composition according to any one of claims 1 to 5 is used to evaluate in vitro efficacy based on a 3D model of vaginal mucosal epithelial cells.

9. The use according to claim 8, characterized in that Methods for evaluating in vitro efficacy based on the 3D model of vaginal epithelial cells include: Human vaginal epithelial cells were seeded on Matrigel or acellular vaginal matrix scaffolds, added to DMEM / F12 medium containing 10% fetal bovine serum, and cultured in a 37°C, 5% CO2 incubator for 7-10 days to form a 3D model of vaginal mucosal epithelial cells; A negative control group, a positive control group, an experimental group, a fermentation product group alone, and a rose component group alone were set up, wherein the negative control group included a culture medium, the positive control group included 1 μg / mL lipopolysaccharide, the experimental group included the vaginal mucosa repair composition according to any one of claims 1 to 5, the fermentation product group alone included a Lactobacillus rhamnosus fermentation product with a solid content of 10%, and the rose component group alone included a rose component with a solid content of 10%; The negative control group, positive control group, experimental group, fermentation product single group, and rose component single group were added to the vaginal mucosal epithelial cell 3D model to detect transepithelial electrical resistance, inflammatory factor secretion, and histological structure; When the transepithelial electrical resistance of the experimental group is ≥ that of the negative control group, and the inflammatory factor secretion content is ≤ that of the negative control group, the vaginal mucosa repair composition has mucosa repair and itching relief effects.

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