TWI666316B - Lactic acid bacteria composition capable of preventing and treating bacterial vaginitis - Google Patents

Abstract

A composition for preventing or treating bacterial vaginosis and regulating vaginal immunity, which has an effective amount and is selected from the group consisting of: Lactobacillus rhamnosus GMNL-680, Lactobacillus plantarum GMNL- The group consisting of 682; the composition of the present invention is used for preventing and treating bacterial vaginosis by inhibiting the growth of pathogenic bacteria and regulating the expression of immunoregulatory factors, and is present in a pharmaceutical composition, a food, or a combination thereof.

Description

Lactic acid bacteria composition capable of preventing and treating bacterial vaginitis

The present invention relates to the use of lactic acid bacteria compositions for the prevention and treatment of bacterial vaginosis and for the regulation of vaginal immune function.

Bacterial vaginosis (BV) is the most common vaginitis. The main cause of illness is the increased pH of the vagina, which leads to an increase in the complexity of the alkaline environment and the increase in non-single pathogen infections (bad bacteria), accompanied by resident lactic acid bacteria ( The number of probiotics is reduced. Common pathogens include Gardner, Escherichia coli, Candida albicans, Candida tropicalis, A. vaginalis, Prevo, Proteus, etc. (Mastromarino P. Et al., New microbiologica , 2013, 36, 229-238).

Symptoms of vaginal discomfort, odor, abnormal secretions, etc., there are two kinds of diagnosis of bacterial vaginitis: (1) Amsel criteria, four of the clinical symptoms meet the following three for diagnosis: vagina The secretion is watery gray and adheres to the vaginal wall; the vaginal pH is greater than 4.5; Whiff test: the secretion is exposed to 10% KOH and emits ammonia odor (fishy smell); under the microscope, a large amount of bacteria adsorption can be seen on the smear sample. In the vaginal epithelial cells, the cells are presented with clue cells in a granular or spotted appearance; (2) according to the Nugent score of the vaginal gram stain, when scoring 7 or a score of 4 to 7 points and the clue cells are visible in the specimen, which is bacterial vaginitis.

In the treatment of bacterial vaginosis, the most commonly used antibiotic drugs for oral or topical use, such as Metronidazole, however, the use of antibiotics will not only kill the pathogens causing BV, but also cause normal vaginal The death of the flora, and the growth of probiotics in the body tends to be slower than that of the pathogens. Therefore, the drug treatment often causes the patient's immunity to decrease, causing the patient's vaginal bacteria to change, thereby repeating the infection and even causing the pathogen to develop resistance. Treatment barriers, in addition, there is a potential risk of side effects. Although the treatment of imidide has a clinically significant effect, it still has a recurrence rate of up to 50%, and the aforementioned disadvantages of using antibiotics are still unavoidable, which is a thorny problem that needs to be overcome in today's treatment, so no side effects are sought. , drug resistance problems, alternative drugs with high efficacy and avoiding recurrence, or treatment options, are important issues related to recent research on bacterial vaginitis (Hongying T. et al. Archives of Gynecology and Obstetrics , 2017, 295, 1131- 1339).

In addition, in the treatment of BV indicators, in addition to the changes in the aforementioned pathogens and probiotics, because the symptoms of inflammation in the affected part involve the activation of immune response, the molecular expression related to immune regulation is also a reference basis, in which IL -10 and IL-12 are immunoregulatory factors that are produced in large quantities when invaded by pathogens, which synergistically promote the host's immune defense and regulate body fluid balance (Ma X. et al., F1000 Research, 2015 , 4 (F1000 Faculty) Rev), 4, 1465), and in addition to IL-10 and IL-12, Dmitry V. et al. in 1996 also elaborated on the role of IL-1β and IL-6 in bacterial infections. Both are pro-inflammatory cytokines, IL-1β promotes phagocytosis of the host's immune cells, and IL-6 assists the neutrophil to clear bacteria in a non-phagocytic manner and regulate the development of inflammatory reactions (Dmitry V.et Al. Int . J. of Antimicrobial Agents , 1996 , 1, p33-40), after administration, the decrease in the amount of expression may represent a mitigation of the inflammatory response in the affected area, implying an improvement in inflammation through the above-mentioned cellular immune regulation. Factor performance change Shape, can be used as indicators of the efficacy of treatment of bacterial infections by reference.

In recent years, the literature pointed out that the use of probiotics (lactic acid bacteria) can help BV patients restore the normal bacteriological balance of the vagina after receiving antibiotics. In a paper published in 2015, Jean et al. proposed a new generation of drugs for the treatment of vaginitis, Tinidazole, which was administered alone or in combination with Lactobacillus (Lactobacillus) in probiotics. ( Lactobacillus reuteri ) RC-14 and Lactobacillus rhamnosus GR-1 were administered to patients. Lactobacillus iners were detected in the vagina of the patient through 16s rRNA in a combination of probiotics. As well as a significant increase in Lactobacillus crispatus , this combination of antibiotics and probiotics helps to restore the balance of the patient's vaginal flora and enhance the chances of avoiding recurrence (Jean M. et al., Microbial Ecology in Health & Disease , 2015;14 ;26:27799); Also in 2015, Piotr B et al. also pointed out that the antibiotic drug Metronidazole was combined with prOVag® (containing Lactobacillus gasseri 57C, Lactobacillus fermentum ) containing three probiotic formulas. 57A and Lactobacillus plantarum 57B) can effectively prolong the recurrence time of patients with bacterial vaginitis, reduce the pH of the vagina, and The number of Lactobacillus vaginalis after treatment with conventional antibiotics.

In view of the aforementioned probiotics combined with antibiotic therapy can improve the chances of avoiding recurrence, antibiotic administration may still cause side effects, drug resistance, and high recurrence rate. Therefore, some studies have tried not to use antibiotics, but to test probiotics alone. Whether the combination of bacteria can treat bacterial vaginitis, the related patents are briefly described below.

Taiwan Patent No. TW I542353 discloses a food composition and/or pharmaceutical composition for combating vaginitis, comprising: at least one strain of lactic acid bacteria having gastric acid, bile salt and mold inhibitor Clotrimazole tolerance, It is selected from Lactobacillus acidophilus F-1 strain, Lactobacillus rhamnosus CT-53 strain or a combination thereof to inhibit tumor necrosis factor-α (TNF-α) and promote cell interleukin The expression of IL-10 (IL-10), which inhibits the symptoms of vaginal mucosal cells, can be administered orally or directly in the form of the affected area. However, it was initially tested in the direction of immunoregulation, and further strains that survived the digestive tract were screened through bile salts, acid resistance and mildew inhibitors. However, the root cause of vaginitis is mainly the increase of pathogenic bacteria and the reduction of probiotics. This invention is only to prove its regulation of immunoregulatory factors and the relief of symptoms, but it does not directly prove its influence on its vaginal flora. There is also no assessment of the human effect applied to the affected part of the patient, and it is even less likely to prove the possibility of overcoming the high recurrence of vaginitis.

Chinese Patent No. CN105263504A discloses a bacterial composition for preventing and/or curing bacterial vaginosis, vaginitis and vaginal infections associated with Escherichia coli pathogens and/or Gardner, the bacterial mixture comprising plants Lactobacillus strain LMGP-21021 (LP01) or Lactobacillus plantarum strain LMG P-21020 (LP02) and bacterial strain Lactobacillus fermentum DSM26955 (LF15) or bacterial strain Lactobacillus fermentum DSM26956 (LF16), the composition inhibits Escherichia coli And the growth of Gardnerella vaginalis and the release of the immunoregulatory factors IL-4 and IL-10, which are prepared together with the tara gum and then administered to the patient as a vaginal tablet. Clinically, the aforementioned tara gum acts as a physical barrier to harmful microorganisms. At 24 hours and 48 hours after administration, it showed the highest inhibitory activity against Gardnerella vaginalis and effectively blocked the acute infection caused by the bacterium. The Lactobacillus composition of the present invention is administered into a vaginal tablet in a mixed tara gum manner, although the Nugent score of the patient can be effectively reduced in human clinical tests. However, an important feature of the invention is that the tara gum can act on the affected part to form The physical barrier to pathogens to avoid acute infections. Therefore, the dosage form needs to be a vaginal tablet or any route that can be directly applied to the affected part. However, since the affected part of the vaginitis patient is more concealed, the vaginal piece is not an instant convenient route of administration. Moreover, because the parts are relatively fragile and inadvertently used, it will cause second-degree injury to the patient. Therefore, how to treat the vaginitis in a manner that is not limited by the dosage form and is more convenient and reduces the probability of recurrence remains a problem to be solved.

In view of this, the inventor of this case has a deep understanding of the shortcomings and shortcomings of the previous case. He is thinking and improving and innovating. After years of painstaking research, he finally succeeded in developing this piece of Lactobacillus rhamnosus GMNL-680, plant. A group of lactic acid bacteria consisting of Lactobacillus plantarum GMNL-682 to prevent and treat bacterial vaginosis and to regulate immunity.

references

1. Paola Mastromarino et al., Bacterial vaginosis: a review on clinical trials with probiotics. New Microbiologica , 36, 229-238, 2013

2. Hongying Tan et al., Effects of metronidazole combined probiotics over metronidazole alone for the treatment of bacterial vaginosis: a meta-analysis of randomized clinical trials. Archives of Gynecology and Obstetrics , 10.1007/s00404-017-4366-0, 2017

3. Ma X, Yan W, Zheng H et al, Regulation of IL-10 and IL-12 production and function in macrophages and dendritic cells. F1000Research , 4(F1000 Faculty Rev), 1465, 2015

4. Dmitry V. Pechkovsky, Michael P. Potapnevb, Oksana M. Zalutskaya, Different patterns of cytokine regulation of phagocytosis and bacterial killing by human neutrophils. International Journal of Antimicrobial Agents , 1, p33-40, 1996.

5. Jean M. Macklaim et al., Changes in vaginal microbiota following antimicrobial and probiotic therapy, Microbial Ecology in Health & Disease , 14;26:27799, 2015,

6. Piotr B. Heczko et al., Supplementation of standard antibiotic therapy with oral probiotics for bacterial vaginosis and aerobic vaginitis: a randomised, double-blind, placebo controlled trial, BMC Women's Health , 3;15:115, 2015

The present invention utilizes a lactic acid bacteria composition for the prevention and treatment of bacterial vaginosis.

For the purpose of the foregoing invention, the present invention is a lactic acid bacteria composition comprising Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum GMNL-682, wherein the composition has prophylactic or therapeutic bacterial properties. Vaginitis and the effect of regulating immunity.

For the purpose of the foregoing invention, the aforementioned Lactobacillus rhamnosus GMNL-680 is deposited at the Taiwan Food Industry Research Institute under the number BCRC 910775, the China Center for Type Culture Collection (CCTCC), and the registration number is CCTCC M 2017766; Lactobacillus plantarum GMNL-682, deposited at the Taiwan Food Industry Research Institute, numbered BCRC 910776, and the China Center for Type Culture Collection (CCTCC), with accession number CCTCC M 2017767.

For the purpose of the foregoing invention, the aforementioned composition is a pharmaceutical composition, a nutritional supplement, a health food or a combination thereof.

For the purposes of the foregoing invention, the composition may further comprise a pharmaceutically acceptable carrier.

For the purposes of the foregoing invention, the aforementioned compositions are administered in the form of solutions, suspensions, emulsions, powders, lozenges, pills, syrups, troches, tablets, chewing gums, thick pastes, and capsules.

For the purpose of the foregoing invention, the composition may further comprise an edible material comprising water, fluid dairy, milk, condensed milk, yogurt, yogurt, Frozen Yogurt, Lactobacillus fermented beverage, milk powder, ice cream, cheese, cheese, soy milk, fermented soy milk, vegetable juice, juice, sports drink, dessert, jelly, candy, baby food, health food, animal feed, Chinese herbal medicine, dietary supplement Product.

For the purpose of the foregoing invention, the present invention provides a use of a lactic acid bacteria composition for preparing a food or a medicament for preventing or treating bacterial vaginosis and regulating vaginal immunity, wherein the lactic acid bacteria composition comprises Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum GMNL-682.

For the purpose of the foregoing invention, the aforementioned use for the prevention and treatment of bacterial vaginosis is through the growth of a pathogen which inhibits bacterial vaginitis.

For the purpose of the foregoing invention, the pathogen of the aforementioned bacterial vaginitis comprises Escherichia coli or Gardnerella vaginalis .

For the purposes of the foregoing invention, the aforementioned use of modulating vaginal immunity is by modulating the performance of immunomodulatory factors.

For the purposes of the foregoing invention, the aforementioned immunomodulatory factors refer to IL-1β and IL-6.

The present invention demonstrates that the composition of the Lactobacillus rhamnosus GMNL-680 and the Lactobacillus plantarum GMNL-682 and the two strains contained in the composition can be inhibited by the inhibition zone test. The pathogen grows and has an additive effect; and has better efficacy than the published Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14 compositions.

Another embodiment of the present invention also describes that the mouse spleen cells and Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum GMNL-682 are individually or co-cultured to enhance the immunomodulatory factor IL- 10, and IL-12, to improve the efficacy of vaginal immunity, and thereby inhibit pathogens, and the lactic acid bacteria composition can stimulate higher levels of immunomodulatory factors than the published lactic acid strains GR-1 and RC-14 , has better efficacy.

In another embodiment of the present invention, it has been confirmed that after taking GMNL-680 and GMNL-682 for one month, the pathogen of Gardnerella vaginalis is significantly reduced from the human vaginal sample, and it is also indicated that GMNL-680 and 682 are taken. It does improve the ability of vaginal flora, can effectively inhibit pathogens, and reduce the expression of pro-inflammatory cytokines, reduce the incidence of local immunity, and indeed improve the symptoms of patients with bacterial vaginitis.

The composition of the invention is safe and has no side effects, and can be applied to slow down the symptoms of bacterial vaginitis, and has no common antibiotics, and can prevent or improve the effect of bacterial vaginitis by inhibiting vaginal pathogens and improving host immunity.

Figure 1A shows the bacteriostatic ability of Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum GMNL-682 against Escherichia coli ; Figure 1B is Lactobacillus rhamnosus GMNL-680, and Lactobacillus plantarum GMNL-682 against Gardnerella vaginalis ; Figure 2 is Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum ( Lactobacillus plantarum ) GMNL-682 combination with the bacteriostatic ability of the disclosed Lactobacillus strain composition (L. rhamnosus GR-1 + L. reuteri RC-14); Figure 3A is Lactobacillus rhamnosus ( Lactobacillus rhamnosus ) GMNL-680 and Lactobacillus plantarum GMNL-682 enhance the expression of mouse immunoregulatory factor IL-10; Figure 3B is Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum) GMNL-682 to enhance the performance of mice immunomodulatory factor of IL-12; FIG. 4A based Lactobacillus rhamnosus (Lactobacillus rhamnosus) GMNL-680 and Lactobacillus (Lactobacillus plant Arum ) GMNL-682 combination with the disclosed Lactobacillus strain composition (L. rhamnosus GR-1 + L. reuteri RC-14) to enhance the expression of mouse immunoregulatory factor IL-10; Figure 4B Lactobacillus rhamnosus GMNL-680 combined with Lactobacillus plantarum GMNL-682 and the disclosed Lactobacillus strain composition (L. rhamnosus GR-1 + L. reuteri RC- 14) Comparison of the expression of mouse immunoregulatory factor IL-12.

The present invention is exemplified by the following examples, but the present invention is not limited by the following examples. The materials used in the present invention are commercially available materials which are readily available, and the Lactobacillus rhamnosus GMNL-680 (hereinafter referred to as GMNL-680) used in the present invention is deposited in Taiwan. Institute of Food Industry, number BCRC 910775, China Type Culture Collection (CCTCC), registration number CCTCC M 2017766; Lactobacillus plantarum GMNL-682 [hereinafter referred to as GMNL-682], deposited in the Taiwan food industry Institute, number BCRC 910776, China Type Culture Collection (CCTCC), registration number CCTCC M 2017767.

Example 1, GMNL-680 and GMNL-682 inhibit Escherichia coli and Gardella

The strains used in this case, GMNL-680 and GMNL-682, are lactic acid bacteria colonies that have been screened and separated by sputum samples from healthy female adults in Taiwan, and identified by Lactobacillus rhamnosus and plant milk. Lactobacillus plantarum .

First, the culture of the strain to be tested is carried out. The part of the experimental strain was inoculated with GMNL-680 and GMNL-682 to 1 ml MRS broth by a frozen tube, and statically cultured at 37 ° C for 20 hours. On the next day, 10 ul of overnight culture solution was added to 1 ml of MRS broth, the same as 37 ° C. After aerobic static incubation for 20 hours, the number of bacteria was estimated by spectrophotometer OD600nm. On the preparation of the pathogenic strain, Escherichia coli was cultured in LB medium at 37 ° C for 16-18 hours, and the bacteria were killed. Gardnerella vaginalis was anaerobic cultured in BHI medium (containing 5% defibrinated sheep blood) at 37 ° C for 16-18 hours, and then the number of bacteria was determined by spectrophotometer OD600nm, and the concentration of the bacteria was uniformly adjusted to 5x10 ⁸ cells/ Ml.

Continue to carry out the inhibition zone test, take 5x10 ⁷ CFU (colony forming unit) pathogen solution evenly applied to the solid medium, and then extrude the hole with 11mm glass tube, add 200μl test bacteria solution to each hole, and culture according to the growth rate of different pathogens. Time; after 24 hours and 48 hours of culture, the results of the inhibition zone test were observed, and the diameter of the inhibition zone was measured using an electronic vernier caliper.

The results showed that GMNL-680 and GMNL-682 have inhibitory circles for both pathogens, and the combination of the two strains has a more additive effect (Fig. 1A , Fig. 1B ), indicating that the composition can inhibit the growth of pathogenic bacteria.

In addition, the present embodiment further compares the inhibition zone with the well-known lactobacillus strain, Lactobacillus rhamnosus GR-1 and Lactobacillus reuteri RC-14, which assist in the treatment of vaginitis (Fig. 2 ), as opposed to The aforementioned well-known strain compositions, the GMNL-680 and GMNL-682 compositions of the present invention have better bacteriostatic effects.

Example 2, GMNL-680 and GMNL-682 regulate local immunovaginal immunity by boosting the immunoregulatory factors IL-10 and IL-12.

To test whether GMNL-680 and GMNL-682 can affect the performance of immunoregulatory factors, this example was co-cultured with mouse spleen cells and assayed for immunomodulatory factors (IL-10 and IL-12) in the supernatant. The content changes. First, the culture of the strain was carried out. The preparation of the experimental strain was the same as in Example 1. First, GMNL-680 and GMNL-682 were inoculated by a freezing tube to 1 ml of MRS broth, and the cells were statically cultured at 37 ° C for 20 hours. On the next day, 10 μl of overnight culture was taken. The bacterial solution was transferred to 1 ml of MRS broth, and the same as that at 37 ° C for 20 hours in aerobic static culture, the number of bacteria was estimated at OD600nm, and the MRS culture medium was replaced with RPMI1640 medium containing 10% fetal bovine serum.

On the other hand, the isolation of mouse spleen cells was purchased from the National Experimental Animal Center for 6-week-old BALB/c mother rats. After one week of feeding in the Jingyue biotechnology animal room, the mice were sacrificed with CO ₂ and the spleen was taken out. After grinding, it was suspended in RPMI1640 medium containing 10% fetal bovine serum to prepare a cell suspension.

The suspended mouse spleen cells were seeded into 96-well plates at a cell volume of ⁴ × ^{10 5} per well, and 4× ^{10 6} CFU or 2×10 ⁷ CFU (Multiplicity of infection, MOI=10 or 50) experimental strain was added at 37 ° C. After aerobic static culture for 48 hours, the supernatant was co-cultured, and the content was analyzed by Mouse IL-10, IL-12 ELISA kit (purchased from BD Biosciences).

The results showed that the co-culture of mouse spleen cells with GMNL-680 or GMNL-682 (MOI=10) significantly improved the anti-inflammatory IL-10 and IL-12 expression (Fig. 3A , 3B ); in addition, GMNL-680 The composition with GMNL-682 stimulates mouse spleen cells to produce higher amounts of IL-10 and IL-12 than the well-known RC-14 and GR-1 compositions described above.

Example 3, GMNL-680 and GMNL-682 can be used to treat bacterial vaginosis to improve local discomfort

In order to observe whether the compound of the present invention is clinically effective, a human clinical experiment (clinical registration number: NCT03116789) was carried out. This example is for a 53-year-old female patient with bacterial vaginitis, taking GMNL-680 and 682 compositions (>1×10 ¹⁰ CFU/day) daily, and observing its effect on vaginitis after one month of use; For the efficacy of bacterial vaginosis, in addition to clinical Nugent score observation, vaginal samples were also collected to quantify the real-time polymerase chain reaction (Q-PCR) to detect pathogens of Gardnerella vaginalis , and immunoregulatory factors. (also known as proinflammatory cytokines) changes in IL-1β and IL-6.

The following is a method for extracting DNA and RNA from a vaginal sample and measuring the amount of expression by Q-PCR. First, the vaginal sample is used to extract DNA and RNA: (1) the part of DNA is taken, and after taking a vaginal sample with a cotton swab, Immerse in PBS and store it in frozen place. Thaw the sample when it is needed for measurement. Extract DNA with DNeasy Blood & Tissue extraction kit (Qiagen). (2) In addition, extract the RNA part by centrifuging the vaginal sample and sedimenting. After adding 1ml of TRIzol; add 0.2ml chloroform upside down 15 times, let it react at room temperature for 5 minutes, then centrifuge; obtain the supernatant, add 0.5ml isopropanol, let stand at room temperature for 10 minutes, then centrifuge; after centrifugation The supernatant was removed, the pellet was washed with 1 ml of 75% alcohol, the precipitate was dried, 20-50 μl of DEPC water was added to dissolve back, and the RNA concentration was determined; (3) RNA was reversely transcribed, and 1-5 μg of the previously extracted sample was taken. RNA supplemented with RNase free water to a total volume of 10 μl, added 10X Random primer (2 μl), 10 mM dNTP (1 μl), applied at 65 ° C for 5 minutes, then placed on ice for 2-3 minutes; then added 4 μl of 5X RT (reverse transcriptase) buffer, 1μl 0.1M DTT, 1μl RNase inhibitor (In In vitro, RNaseOUTTM), 1 μl of RT enzyme (Invitrogen, SuperScript® III), mixed for 5 minutes at room temperature, then placed at 50 ° C for 60 minutes, and at 70 ° C for 15 minutes to complete reverse transcription.

The amount of expression was determined by Q-PCR, and the previously extracted DNA or the reverse-transcribed cDNA was diluted with sterile water, and 2 μl was taken, and Q was performed with 2X Rotor-Gene SYBR Green PCR Master Mix (Qiagen, Cat. 204076). -PCR reaction; the results of the analysis are expressed by 2 ^{- △ CT} (2- ^{△ CT = 2-(CT target gene-CT GAPDH)} or 2 ^{- (CT GV-CT total bacterial)} ), △ is the relative expression amount, which is deduction The higher the value of GAPDH or total bacterial, the higher the expression, the higher the expression, the results are shown in Table 1 ; 53-year-old women with bacterial vaginitis, taking GMNL-680 and 682 compositions daily (> 1 × 10 ¹⁰ CFU / day One month later, the Nugent score for the evaluation of vaginitis decreased significantly, and the vaginal pathogen Gardnerella vaginalis expression, immunoregulatory factors (proinflammatory cytokines IL-1β and IL-6) also showed a significant decrease, indicating that the composition It can effectively inhibit vaginal pathogens and reduce the incidence of vaginal local immune response in women, thereby improving the discomfort and severity of vaginitis.

The invention proves that GMNL-680 or GMNL-682 and the combination of the two strains can inhibit the growth of vaginal pathogens, and the two strains have additive effects; in addition, the mouse spleen cells are cultured separately with GMNL-680 or GMNL-682. It can stimulate the spleen cells to secrete cytokines, enhance the expression of IL-10 and IL-12, and achieve the effect of immune regulation, and whether it inhibits pathogens or enhances the release of immune regulatory factors, compared with the published rhamnosylose Bacillus sp. GR-1 and L. reuteri RC-14 compositions have better efficacy; in addition, after a month of administration of the composition of the present invention, the Nugent score of the affected part of the vaginitis patient is significantly reduced, and the vaginal pathogen is passed through human clinical trials. The performance of Gardnerella vaginalis , the pro-inflammatory cytokines IL- 1β and IL-6 also decreased significantly, indicating that taking GMNL-680 and 682 can be used to treat bacterial vaginosis and regulate vaginal immune regulators to treat vaginitis and soothe Discomfort.

The lactic acid bacteria composition is a safe and no side effect of Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum GMNL-682 composition, which regulates the expression of immunoregulators to enhance vaginal immunity, thereby It can slow down the symptoms of bacterial vaginitis and has better curative effect than the published Lactobacillus composition, and there is no worry that the normal bacteria in the vagina will die due to the use of antibiotics, and the bacteria phase will change, and then the infection will be repeated. The potential drug resistance problem and the risk of side effects of the drug, the present invention can simultaneously prevent or improve the effect of bacterial vaginitis by inhibiting the vaginal pathogen and improving the local vaginal immunity of the host.

Therefore, this case is not only innovative in terms of technical thinking, but also has many functions that are not in the traditional methods of the past. It has fully complied with the statutory invention patent requirements of novelty and progressiveness, and applied for it according to law. You are requested to approve the patent application for this invention. The case, in order to invent, to the sense of virtue.

【Biomaterial Storage】

Domestic registration information [please note according to the registration authority, date, number order]

1. Depository institution - Food Industry Development Research Institute, storage date - April 14, 2017, registration number - BCRC 910775

2. Depository institution - Food Industry Development Research Institute, storage date - April 14, 2017, registration number - BCRC 910776

Foreign deposit information [please note according to the country, organization, date, number order]

1. Host Country - China, Hosting Organization - China Type Culture Collection (CCTCC), Hosting Date - November 3, 2017, Registration Mark - CCTCC M 2017766

2. Host Country - China, Hosting Organization - China Type Culture Collection (CCTCC), Hosting Date - November 3, 2017, Registration Mark - CCTCC M 2017767

Claims (10)

A lactic acid bacteria composition comprising Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum GMNL-682, wherein the composition has the effect of preventing or treating bacterial vaginosis and regulating vaginal immunity; The registration number of the Lactobacillus rhamnosus GMNL-680 is BCRC 910775, and the registration number of Lactobacillus plantarum GMNL-682 is BCRC 910776. The composition of claim 1 , wherein the composition is a pharmaceutical composition, a nutritional supplement, a health food, or a combination thereof. The composition of claim 2 , wherein the composition further comprises a pharmaceutically acceptable carrier. The composition of claim 2 , wherein the composition is administered in the form of a solution, a suspension, an emulsion, a powder, a lozenge, a pill, a syrup, a lozenge, a tablet, a chewing gum. , thick paste, capsules, vaginal suppositories or topical agents. The composition of claim 2 , wherein the composition further comprises an edible material comprising water, fluid dairy, milk, concentrated milk, yogurt, yogurt, frozen yogurt Lactobacillus fermented beverage, milk powder, ice cream, cheese, cheese, soy milk, fermented soy milk, vegetable juice, juice, sports drink, dessert, jelly, candy, baby food, health food, animal feed, Chinese herbal medicine or dietary supplement. A lactic acid bacteria composition for preparing a medicament for preventing or treating bacterial vaginosis and regulating vaginal immunity, wherein the lactic acid bacteria composition comprises Lactobacillus rhamnosus GMNL-680 and Lactobacillus plantarum GMNL -682; wherein the registration number of the Lactobacillus rhamnosus GMNL-680 is BCRC 910775, and the registration number of Lactobacillus plantarum GMNL-682 is BCRC 910776. The use of the prophylactic or bactericidal vaginitis refers to the inhibition of the growth of pathogenic bacteria of bacterial vaginitis, as claimed in claim 6 . The use of the bacterial vaginitis refers to Escherichia coli or Gardnerella vaginalis , as described in claim 7 . For use as described in claim 6 , the use of modulating vaginal immunity is by modulating the performance of immunomodulatory factors. The use of the immunomodulatory factor according to the invention of claim 9 is IL-1β and IL-6.