Use of traditional chinese medicine compound composition in preparing drug for treating inflammations in female reproductive tract

Abstract

Disclosed in the present application is the use of a traditional Chinese medicine compound composition in preparing a drug for treating inflammations in the female reproductive tract. The traditional Chinese medicine compound composition comprises 50 parts of phellodendri amurensis cortex, 50 parts of cnidii fructus, 30 parts of sophorae flavescentis radix, 20 parts of catechu, 10 parts of dried alum, and 8 parts of coptidis rhizoma. In the use disclosed in the present application, on the basis of the syndrome differentiation in traditional Chinese medicine, the traditional Chinese medicine compound composition for external use having the effects of clearing heat, drying dampness and removing toxins is formulated. The traditional Chinese medicine compound composition not only possesses an inhibitory effect on various pathogenic microorganisms in the female reproductive tract, but also can keep the activity of probiotics in the female reproductive tract compared with antibiotics, thereby achieving a better therapeutic effect on restoring a healthy microecology of the reproductive tract.

Description

Application of a traditional Chinese medicine compound composition in preparation of a medicine for treating inflammation of female genital tract

[0001] The present application claims priority to the Chinese patent application No. 2024107215050 filed on June 04, 2024, and entitled "Application of a traditional Chinese medicine compound composition in preparation of a medicine for treating inflammation of female genital tract", the content of which is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0002] The present application relates to the technical field of pathogenic microorganisms, in particular, the present application relates to the application of a traditional Chinese medicine compound composition in preparation of a medicine for treating inflammation of female genital tract. BACKGROUND

[0003] The existing treatment for inflammation of female genital tract mainly relies on antibiotics, but the antibiotic resistance is becoming increasingly serious, and the local microenvironment is easily destroyed, the recurrence rate is high after treatment, and the long-term efficacy is not ideal. At the same time, the inflammation of female genital tract is often complicated with multiple pathogen infections, and the use of single antibiotic often has poor curative effect. The inventors realize that traditional Chinese medicine can play a unique therapeutic advantage through the effects of clearing heat, drying dampness and detoxification. SUMMARY

[0004] The present application proposes the application of a traditional Chinese medicine compound composition in preparation of a medicine for treating inflammation of female genital tract to solve the technical problems of obvious drug resistance, short-term efficacy and large side effects in the related art.

[0005] The present application provides the application of a traditional Chinese medicine compound composition in preparation of a medicine for treating inflammation of female genital tract, wherein the traditional Chinese medicine compound composition comprises 50 parts of Cortex Phellodendri, 50 parts of Fructus Cnidii, 30 parts of Sophora flavescens, 20 parts of Folium Uncariae, 10 parts of Potassium Sulphate and 8 parts of Rhizoma Coptidis.

[0006] Alternatively, the medicine comprises an extract of the traditional Chinese medicine compound composition, and the preparation of the extract comprises the following steps:

[0007] The traditional Chinese medicine compound composition is prepared;

[0008] After being fully soaked with 10-12 times of water, the medicine is decocted with strong fire to obtain a first filtrate;

[0009] The residue is repeatedly decocted to obtain a second filtrate;

[0010] The first filtrate and the second filtrate are combined to become the extract stock solution.

[0011] Further alternatively, the extract stock solution is prepared into a decoction, a concentrated solution, a gel or a dry powder.

[0012] Further, the medicine is an external medicine.

[0013] Alternatively, the method of using the medicine comprises sitz bath or vaginal douche at the concentration of the extract stock solution.

[0014] Further alternatively, the method of using the medicine comprises once a day for several days.

[0015] Further, the medicine is capable of inhibiting the growth of at least one of the following female reproductive tract pathogenic microorganisms: Micrococcus flavus, Prevotella, Atopobium, Escherichia coli, Staphylococcus aureus, Candida albicans, Streptococcus group A, Streptococcus group B, Pneumococcus, and Gardnerella within 20 minutes of administration.

[0016] Further, the medicine is capable of maintaining the activity of female reproductive tract probiotics within 24 hours of administration.

[0017] Alternatively, the female reproductive tract probiotics comprise Lactobacillus crispatus, Lactobacillus gasseri, and Lactobacillus jensenii.

[0018] Further, the medicine improves at least one of the following clinical symptoms: pruritus of the vulva, odor of leucorrhea, excessive amount of leucorrhea, dyspareunia, lower abdominal pain, frequent urination, urgency, pain, anal distension, abnormal vaginal discharge, vaginal inflammation, and vaginal flora imbalance. The technical solutions provided in the embodiments of the present application have the following beneficial technical effects:

[0019] (1) The application, in combination with the method of TCM syndrome differentiation, prepares an external TCM compound composition with the effects of clearing heat, drying dampness, and detoxifying. The TCM compound composition has inhibitory effects on a variety of female reproductive tract pathogenic microorganisms, and can maintain the activity of female reproductive tract probiotics, and has a good curative effect on restoring the healthy microecology of the reproductive tract, as compared with antibiotics.

[0020] (2) The application, in which the TCM compound composition has weaker inhibitory effects on female reproductive tract probiotics than antibiotics with comparable effects, can better protect the balance of female reproductive tract flora, reduce the side effects of the medicine, and reduce the occurrence of repeated attacks of the disease.

[0021] Additional aspects and advantages of the application will be set forth in part in the description which follows, and in part will become apparent to those skilled in the art upon examination of the following and / or can be learned by practice of the application. BRIEF DESCRIPTION OF DRAWINGS

[0022] The above and / or additional aspects and advantages of the application will become apparent and be readily understood by considering the following detailed description, from which the above-mentioned and other aspects and advantages of the application will become apparent, and which, taken in conjunction with the accompanying drawings, illustrate the embodiments of the application.

[0023] Figure 1 is the inhibitory effect of different concentrations of the traditional Chinese medicine compound composition of the present application on Micrococcus flavus, Prevotella and Atopobium;

[0024] Figure 2 is the inhibitory effect of the traditional Chinese medicine compound composition of the present application on Candida albicans;

[0025] Figure 3 is the inhibitory effect of the traditional Chinese medicine compound composition of the present application on Gardnerella;

[0026] Figure 4 is the inhibitory effect of the traditional Chinese medicine compound composition of the present application on Megasphaera;

[0027] Figure 5 is the inhibitory effect of the traditional Chinese medicine compound composition of the present application on Group A Streptococcus;

[0028] Figure 6 is the inhibitory effect of the traditional Chinese medicine compound composition of the present application on Group B Streptococcus;

[0029] Figure 7 is the inhibitory effect of the traditional Chinese medicine compound composition of the present application on Lactobacillus crispatus, a vaginal probiotic. DETAILED DESCRIPTION

[0030] The embodiments of the present application will be described below in conjunction with the accompanying drawings. It should be understood that the embodiments described below in conjunction with the accompanying drawings are exemplary descriptions of the technical solutions of the present application, and do not limit the technical solutions of the present application.

[0031] It can be understood by those skilled in the art that, unless specifically stated, "the" and "that" used herein can also include plural forms. It should be further understood that the phrase "comprising" used in the specification of the present application means that the features, integers, steps, operations, etc. are present, but does not exclude other features, information, data, steps, operations, elements, components, and / or combinations thereof supported by the present art. The term "and / or" used herein means at least one of the items defined by the term, for example, "A and / or B" can be implemented as "A", or as "B", or as "A and B".

[0032] To make the purpose, technical solutions and advantages of the present application clearer, the embodiments of the present application will be described in further detail below in conjunction with the accompanying drawings.

[0033] Reproductive tract inflammation is a common disease that plagues women of childbearing age, including various disease types such as bacterial vaginosis (BV), vulvovaginal candidiasis (VVC), aerobic vaginitis (AV) and mycoplasma infection, mainly with pruritus of vulva, abnormal vaginal discharge and other clinical manifestations. Reproductive tract inflammation is associated with many complications, including pelvic inflammatory disease, infertility, premature delivery, premature rupture of membranes and other adverse pregnancy outcomes, which seriously affect the reproductive health and quality of life of women of childbearing age.

[0034] The female reproductive tract microecology is centered on the vaginal microbiome (VMB), and the interaction between bacterial genera and the maintenance of a certain balance are important prerequisites for female reproductive health. In a broad sense, the connotation of reproductive tract microecological balance is the balance of synergy, antagonism and restraint between different bacterial groups in the reproductive tract. Currently, it is believed that the typical normal reproductive tract bacterial community is dominated by Lactobacillus, which plays an important role in maintaining the host's reproductive health. In addition to Lactobacillus, Streptococcus, Megasphaera, Gardneralla, Escherichia coli, Prevotella and other bacteria, as well as viruses, mycoplasma, chlamydia and Candida albicans, together constitute the reproductive tract microbiome. BV is usually defined as a multi-bacterial state, characterized by the enrichment of gram-negative anaerobes associated with increased risk of disease. VVC is mainly caused by Candida albicans infection and is the second largest reproductive tract inflammation after BV. The characteristics of AV are the decrease in the number of Lactobacillus and the increase in the number of aerobic pathogens. Other sexually transmitted diseases (STDs) also show an imbalance in the reproductive tract microecology, such as the close association between the transmission of chlamydia and the reproductive tract of inert Lactobacillus. Although different types of changes in the reproductive tract microbiota are found in different populations with reproductive tract inflammation, this change is often manifested in microcosm as the increase or depletion of certain specific bacteria, but in macrocosm, its essence can be summarized as 'balance broken', therefore, the treatment principles for different reproductive tract inflammations are the same, i.e. to restore the balance of reproductive tract microecology.

[0035] The current treatment of female genital tract inflammation is still the first choice of antibiotics. Antibiotic resistance. For the treatment of BV, the guidelines recommend metronidazole as the first choice of drug, and consider that metronidazole can inhibit the growth of anaerobic bacteria and has little effect on lactobacillus, which is a relatively ideal treatment drug. However, in the presence of "ideal drug" treatment, the recurrence rate of BV 1 month after treatment can be as high as 20%, 3 months after treatment can be as high as 40%, and 12 months after treatment can be as high as 60%. Fluconazole is the main drug for the treatment of VVC, which can effectively control the infection during oral maintenance therapy, but only about half of the patients can achieve long-term remission after drug withdrawal, and the recurrence rate is as high as 21% ~ 57.1%. At the same time, the incidence of AV is increasing, which has become a global health problem. Reports show that its incidence varies from 5% to 80% due to differences in demographic factors such as race, geographical location and socioeconomic status. The high incidence of AV worldwide is closely related to the emergence of antibiotic resistance, and this situation highlights the urgency of seeking new treatment strategies for AV. Therefore, for the treatment of genital tract inflammation, it is urgent to develop long-term effective, safe and comprehensive treatment methods.

[0036] Oral and topical use of antibiotics is still the main means of treating genital tract inflammation. However, antibiotic therapy has the following disadvantages: (1) antibiotic resistance: bacteria can develop resistance to antibiotics through different ways, such as changing structure, avoiding binding with antibacterial drugs; producing various enzymes to destroy the structure of antibacterial drugs; building their own defense system, closing the binding channel of antibacterial drugs or excluding antibacterial drugs that have entered the bacterial body outside the body, etc. (2) can destroy the beneficial bacteria in the genital tract: the drug action of antibiotics usually targets the specific biological function of a class of microorganisms, so the use of antibiotics not only has the effect of killing pathogenic microorganisms, but also can affect the growth of some beneficial bacteria, and even cause local microenvironment imbalance, easy to cause inflammation for a long time or repeated attacks. (3) Genital tract inflammation is often easy to combine with multiple pathogenic microorganism infections, and one antibiotic only targets one disease, with narrow applicability.

[0037] Traditional Chinese medicine classifies genital tract inflammation as "leukorrhea disease", and believes that "if the body is affected by external factors and the internal organs are damaged by emotions, it will develop into a disease, causing the belt vein to relax and unable to control the various veins, thus there are things in the yin, and it is called leukorrhea." Therefore, "leukorrhea is wet", which contains both deficiency and excess: on the one hand, it is affected by dampness, and contains dampness and heat or heat and toxin, on the other hand, it is caused by dysfunction of internal organs, which can be initially corresponding to pathogen infection and immune dysfunction. The six evils are the main pathogenic factors of exogenous diseases and important conditions for the occurrence of diseases. From the perspective of modern etiology, biological pathogenic factors such as bacteria and viruses belong to the category of six evils. Therefore, affected by dampness, containing dampness and heat or heat and toxin, wet, hot and toxic are the causes of leukorrhea disease. Based on the theory of etiology combined with the view of ecology, female genital tract inflammation and external dampness, internal dampness and heat, and dampness and heat are the same, and wet, hot and toxic are the causes of genital tract inflammation. Clearing heat, drying dampness and detoxifying are the basic methods for treating genital tract inflammation. Traditional Chinese medicine external treatment belongs to the treatment of "leukorrhea disease" and has strong disinfection and insecticidal effects, is convenient to carry and store, and has broad market application prospects.

[0038] The purpose of the present application is to provide the application of a traditional Chinese medicine compound composition with the effect of clearing heat, drying dampness and detoxifying in the preparation of a drug for treating female genital tract inflammation.

[0039] (1) Traditional Chinese medicine compound composition and its preparation process

[0040] The present application provides a traditional Chinese medicine compound composition for treating genital tract inflammation, which is prepared from the following components by weight: 50 parts of Huang Bai, 50 parts of Shechuangzi, 30 parts of Kuqian, 20 parts of Echa, and 10 parts of Huanglian. The specific preparation steps include:

[0041] ① According to the original prescription ratio of 50:50:30:20:10, the medicinal materials are weighed, and 10 times (volume ratio) of pure water (1.5 L) is soaked for 1 h;

[0042] ② Boil for 1 h;

[0043] ③ Filter with gauze and collect the first filtrate;

[0044] ④ Add 1.2 L of pure water again and re-boil for 1 h;

[0045] ⑤ Filter with gauze, collect the second filtrate, and combine the two filtrates to become the original liquid extract;

[0046] ⑥ After slightly concentrating with a decoction pot, use a rotary evaporator to concentrate to 0.4 L for storage;

[0047] In use, the above 0.4L concentrated solution is diluted with warm water to 1.5L for use, so that the use concentration is restored to the concentration of the extract stock solution. Further, when the extract stock solution is made into semi-finished or finished products for sale, the extract stock solution can be packaged as a decoction, or the extract stock solution is prepared into a concentrated solution, or further a coagulant is added to the concentrated solution to make an external use gel product, or the concentrated solution is further concentrated and dried to make a dry powder. The foregoing concentrated solution, gel or dry powder all need to be restored to the concentration of the extract stock solution for use.

[0048] Further, the amount of water added during the two decoctions in the preparation of the extract stock solution can be controlled within the range of 10-12 times by volume, and the concentration of the extract stock solution obtained after decoction can fluctuate within a certain range, which can be adjusted to the appropriate concentration by later concentration or dilution.

[0049] (2) In vitro antibacterial experiment

[0050] The test bacteria (Candida albicans, Streptococcus group A, Streptococcus group B, Micrococcus, Gardnerella, Atopobium, Prevotella, Staphylococcus aureus, Escherichia coli, Lactobacillus crispatus) were washed with PBS to prepare a bacterial suspension (the recovery concentration was about 1-9 x 10^4 CFU / mL). Prepare traditional Chinese medicine and control sample solution, divided into three groups: traditional Chinese medicine compound composition (set multiple concentrations), antibiotic 1 mg / mL (use different antibiotics), culture medium control. Take the above bacterial suspension, respectively, add 100 uL of bacterial solution to the traditional Chinese medicine compound composition, antibiotic and culture medium sample solution, mix evenly, after 20 min and 24 h, take 0.5 mL sample solution into a test tube containing 5 mL PBS, mix thoroughly. Take 0.5 mL and place it in two plates, use 15 mL of nutrient agar medium or sabouraud agar medium cooled to 40-45℃ for pouring, rotate the plate to make it evenly distributed. After the agar solidifies, turn the plate over, incubate at 35℃±2℃ for 48 h or 72 h, count the number of colonies. The culture medium is used as a negative control, and the antibiotic is used as a positive control to evaluate the inhibition rate of the traditional Chinese medicine compound composition on different bacteria.

[0051] ①Effect of traditional Chinese medicine compound composition on Micrococcus (M), Prevotella (P) and Atopobium (A)

[0052] Referring to FIG. 1, Compound 1 is a traditional Chinese medicine compound composition containing crude drug concentration of 31.7 mg / mL; Compound 2 is a traditional Chinese medicine compound composition containing crude drug concentration of 21.12 mg / mL. Evaluate the number of colonies after different traditional Chinese medicine compound compositions with crude drug concentration intervention for 12 h (A), 18 h (B) and 24 h (C). The higher the number of colonies, the more the number of surviving bacteria, and the weaker the antibacterial effect. The results show that the traditional Chinese medicine compound composition significantly inhibits Micrococcus (M), Prevotella (P) and Atopobium (A).

[0053] Effect of Chinese medicine compound composition on E. coli and S. aureus

[0054] Referring to Table 1 and Table 2, the compound concentration 1 is the Chinese medicine compound composition containing the crude drug concentration of 100 mg / mL; the compound concentration 2 is the Chinese medicine compound composition containing the crude drug concentration of 200 mg / mL; the compound concentration 3 is the Chinese medicine compound composition containing the crude drug concentration of 400 mg / mL; the values in the table are the colony numbers (number), and the higher the value, the higher the survival number of the bacteria, and the weaker the inhibition effect. The antibacterial effect is evaluated after the Chinese medicine compound composition is intervened for 2 min, 5 min, 10 min and 20 min. It is found that the Chinese medicine compound composition with different concentrations has a certain inhibitory effect on E. coli and S. aureus after a short time of intervention.

[0055] Table 1 Inhibitory effect of different concentrations of Chinese medicine compound composition on E. coli in a short time

[0056] Table 2 Inhibitory effect of different concentrations of Chinese medicine compound composition on S. aureus in a short time

[0057] Effect of Chinese medicine compound composition on C. albicans

[0058] Referring to FIG. 2, the Chinese medicine compound composition contains the crude drug concentration of 112 mg / mL, and the control antibiotic is fluconazole 0.6 mg / mL; the horizontal coordinate of the column chart is the colony number (number), and the higher the value, the higher the survival number of the bacteria, and the weaker the inhibition effect. Compared with the negative control group (culture medium), the Chinese medicine compound composition can significantly inhibit the growth of C. albicans after 20 min and 24 h of intervention, and the inhibitory effect of the Chinese medicine compound composition on C. albicans in a short time is comparable to that of the antibiotic.

[0059] Effect of Chinese medicine compound composition on G. vallis (G.V)

[0060] Referring to FIG. 3, the inhibitory effect of the Chinese medicine compound composition is evaluated compared with the positive control (antibiotic) and the blank control (culture medium) after 20 min (A) and 24 h (B) of intervention. The Chinese medicine compound composition contains the crude drug concentration of 112 mg / mL, and the control antibiotic is metronidazole 14 mg / mL; the horizontal coordinate of the column chart is the colony number (number), and the higher the value, the higher the survival number of the bacteria, and the weaker the inhibition effect. The inhibitory effect of the Chinese medicine compound composition on G. vallis after 20 min of intervention is significantly due to the antibiotic, and the inhibitory effect of the Chinese medicine compound composition after 24 h of intervention is comparable to that of the antibiotic, which significantly inhibits the growth of G. vallis.

[0061] Effect of Chinese medicine compound composition on M. cerebralis

[0062] Reference to Figure 4, evaluate the inhibitory effect of the intervention 20 min (A) and 24 h (B) of the traditional Chinese medicine compound composition compared with the positive control (antibiotics), blank control (culture medium). Traditional Chinese medicine compound composition contains crude drug concentration of 112 mg / mL, the control antibiotic is metronidazole 14 mg / mL; the horizontal coordinate of the column chart is the number of colonies (number), the higher the value, the higher the number of surviving bacteria, the weaker the inhibition. After 20 min of intervention, the inhibitory effect of the traditional Chinese medicine compound composition on the macrococcus was slightly better than that of the antibiotic, and after 24 h of intervention, the inhibitory effect of the traditional Chinese medicine compound composition was equivalent to that of the antibiotic, which significantly inhibited the growth of the macrococcus.

[0063] (6) Effect of traditional Chinese medicine compound composition on group A streptococcus

[0064] Reference to Figure 5, evaluate the inhibitory effect of the intervention 20 min (A) and 24 h (B) of the traditional Chinese medicine compound composition compared with the positive control (antibiotics), blank control (culture medium). Traditional Chinese medicine compound composition contains crude drug concentration of 112 mg / mL, the control antibiotic is metronidazole 14 mg / mL; the horizontal coordinate of the column chart is the number of colonies (number), the higher the value, the higher the number of surviving bacteria, the weaker the inhibition. After 20 min of intervention, the inhibitory effect of the traditional Chinese medicine compound composition on the group A streptococcus was slightly better than that of the antibiotic, and after 24 h of intervention, the traditional Chinese medicine compound composition could significantly inhibit the growth of the group A streptococcus.

[0065] (7) Effect of traditional Chinese medicine compound composition on group B streptococcus

[0066] Reference to Figure 6, evaluate the inhibitory effect of the intervention 20 min (A) and 24 h (B) of the traditional Chinese medicine compound composition compared with the positive control (antibiotics), blank control (culture medium). Traditional Chinese medicine compound composition contains crude drug concentration of 112 mg / mL, the control antibiotic is metronidazole 14 mg / mL; the horizontal coordinate of the column chart is the number of colonies (number), the higher the value, the higher the number of surviving bacteria, the weaker the inhibition. After 20 min of intervention, the inhibitory effect of the traditional Chinese medicine compound composition on the group B streptococcus was slightly better than that of the antibiotic, and after 24 h of intervention, the inhibitory effect of the traditional Chinese medicine compound composition was equivalent to that of the antibiotic, which significantly inhibited the growth of the group B streptococcus.

[0067] (8) Effect of traditional Chinese medicine compound composition on Lactobacillus crispatus (L.C)

[0068] Referring to FIG. 7, the bacteriostatic effect of the traditional Chinese medicine compound composition compared with the positive control (antibiotic) and the blank control (culture medium) was evaluated after 20 min (A) and 24 h (B) of intervention. The traditional Chinese medicine compound composition contained 112 mg / mL of crude drug concentration, and the control antibiotic was metronidazole 14 mg / mL; the horizontal coordinate of the column chart was the number of colonies (number), and the higher the value, the higher the number of surviving bacteria, and the weaker the inhibition. After 20 min of intervention, the traditional Chinese medicine compound composition and the antibiotic had no obvious inhibitory effect on the vaginal probiotic Lactobacillus crispatus, and after 24 h of intervention, the antibiotic significantly inhibited the growth of L. crispatus, and the traditional Chinese medicine compound composition slightly inhibited L. crispatus, but the inhibitory effect was significantly worse than that of the antibiotic, indicating that the traditional Chinese medicine compound composition was worse than the antibiotic in terms of the destruction of vaginal probiotics.

[0069] (9) Effect of the traditional Chinese medicine compound composition on L. gasseri (L.G) and L. jensenii (L.J)

[0070] Referring to Tables 3 and 4, the vaginal probiotics L. gasseri and L. jensenii were verified according to the experimental method described above, and the bacteriostatic effect of the traditional Chinese medicine compound composition compared with the positive control (antibiotic) and the blank control (culture medium) was evaluated after 20 min and 24 h of intervention. The traditional Chinese medicine compound composition contained 112 mg / mL of crude drug concentration, and the control antibiotic was metronidazole 14 mg / mL, and the results showed that after 20 min of intervention, the traditional Chinese medicine compound composition and the antibiotic had no obvious inhibitory effect on the vaginal probiotics L. gasseri and L. jensenii, and after 24 h of intervention, the antibiotic significantly inhibited the growth of the two probiotics, and the traditional Chinese medicine compound composition inhibited L. gasseri and L. jensenii, but was significantly worse than the antibiotic, indicating that the traditional Chinese medicine compound composition was significantly worse than the antibiotic in terms of the destruction of the two vaginal probiotics, and the results were consistent with the trend of L. crispatus.

[0071] Table 3 Inhibitory effect of the traditional Chinese medicine compound composition on the vaginal probiotic L. gasseri

[0072] Table 4 Inhibitory effect of the traditional Chinese medicine compound composition on the vaginal probiotic L. jensenii

[0073] (3) Safety experiment

[0074] H8 cells and VK2 / E6E7 cells in logarithmic growth phase were taken and inoculated in 96-well plates at 2x10^4 cells per well after digestion. When the cells were adherent and grew to 80%, 5uL of traditional Chinese medicine compound composition, 5uL of metronidazole solution, 5uL of fluconazole solution, and 5uL of clarithromycin solution were added, and controls were set up, with 3 replicate wells in each group. They were cultured in a 37°C CO2 incubator, and 10% CCK8 solution was added after 12h, 18h, and 24h, respectively. After 4h of reaction in the incubator, the absorbance of each well was detected at A450nm wavelength. Traditional Chinese medicine and different antibiotics had no significant effect on the growth of H8 cells and VK2 / E6E7 cells.

[0075] H8 cells and VK2 / E6E7 cells in logarithmic growth phase were taken and inoculated in 6-well plates at 2x10^5 cells per well after digestion. When the cells were adherent and grew to 80%, drug intervention was performed, and they were divided into five groups: ① traditional Chinese medicine compound composition, ② metronidazole group, ③ fluconazole group, ④ clarithromycin group, and ⑤ control group, with 3 replicate wells in each group. After 24h of culture, the cells were trypsinized, dropped on cover slips, fixed with methanol, and subjected to HE staining to observe cell morphology. Meanwhile, the whole protein of each well after 24h of culture was extracted for routine WB detection, with Caspase3 and Caspase8 proteins as detection indexes and GAPDH as an internal reference. Traditional Chinese medicine and different antibiotics had no significant promoting effect on the apoptosis of H8 cells and VK2 / E6E7 cells.

[0076] (4) Clinical research

[0077] Further, a small-sample self-control clinical trial was conducted to determine the effect of traditional Chinese medicine compound composition on the symptoms and signs of reproductive tract infection. Participants who met the following conditions were included: ① 18-45 year-old women with symptoms of vulvar itching, discomfort, or burning, dyspareunia, and changes in color, quantity, and quality of vaginal discharge; ② with a history of sexual life, non-menopausal, and non-pregnant; ③ no sexual life or vaginal medication or irrigation within 3 days before sampling; at the same time, asymptomatic normal women were used as controls. Clinical information, symptoms and signs, and vaginal secretion samples of participants were collected for 16S rRNA sequencing, detection of various reproductive tract pathogenic microorganisms, routine detection of leukorrhea, and cytokine detection. Participants were given traditional Chinese medicine composition for sitz bath and vaginal irrigation, 1 time / d, for 7 consecutive days. After 7 days, clinical information, symptoms and signs, and vaginal secretion of participants were collected, and the detection indexes were the same as above. The improvement of symptoms, routine indexes of leukorrhea, cytokine indexes, reproductive tract pathogenic microorganisms, and reproductive tract flora composition of symptomatic women and control women before and after treatment with traditional Chinese medicine were compared.

[0078] The results refer to Tables 5-7, and it is found that the Chinese medicine compound composition significantly improves the clinical symptoms of pruritus of the vulva (effective rate 100%), odor of leucorrhea (effective rate 100%), large amount of leucorrhea (effective rate 100%), dyspareunia (effective rate 50%), and anal distending sensation (effective rate 67%), and has certain improvement effect on abnormal vaginal discharge.

[0079] Table 5 Comparison of clinical symptoms before and after treatment with the Chinese medicine compound composition

[0080] Table 6 Comparison of gynecological examination before and after treatment with the Chinese medicine compound composition

[0081] Table 7 Comparison of vaginal discharge examination before and after treatment with the Chinese medicine compound composition

[0082] In summary, the Chinese medicine compound composition of the present application has the following advantages when applied to the treatment of female reproductive tract inflammation: (1) The Chinese medicine compound composition of the present application is derived from natural herbs, has small side effects, is safer than antibiotics, and has wide applicability. (2) It can inhibit a variety of common reproductive tract pathogenic microorganisms, such as Candida albicans, Streptococcus group A, Streptococcus group B, macrococcus, Gardnerella vaginalis, Atopobium, Prevotella, etc. Compared with existing drugs, it has the characteristics of wide application and better efficacy, and has significant application value. (3) In vitro bacteriostatic experiments show that the Chinese medicine compound composition of the present application has less inhibitory effect on vaginal probiotic Lactobacillus crispatus than antibiotics, which is conducive to the recovery and maintenance of the healthy environment of the female reproductive tract dominated by Lactobacillus crispatus.

[0083] In summary, the present application provides a Chinese medicine compound composition for treating female reproductive tract inflammation, wherein the Chinese medicine compound composition comprises 50 parts of Cortex Phellodendri, 50 parts of Cnidium, 30 parts of Sophora, 20 parts of Folium Uncariae, 10 parts of Potassium Sulphate, and 8 parts of Rhizoma Coptidis. The medicine comprises an extract of the Chinese medicine compound composition, and the preparation of the extract comprises the following steps:

[0084] The Chinese medicine compound composition is prepared, and after being soaked with 10-12 times the volume of water, it is boiled with a strong fire to obtain a first filtrate. The residue is repeatedly boiled to obtain a second filtrate. The first filtrate and the second filtrate are combined to obtain the extract stock solution.

[0085] The leaching solution is prepared into a decoction, a concentrated solution, a gel or a dry powder. The medicine is an external medicine. The method for using the medicine comprises sitz bath, external genitalia washing or vaginal irrigation with the concentration of the leaching solution. The method for using the medicine comprises once a day, and continuous administration for several days. The medicine can inhibit the growth of at least one of the following female reproductive tract pathogenic microorganisms within 20 minutes after administration: Micrococcus flavus, Prevotella, Atopobium, Escherichia coli, Staphylococcus aureus, Candida albicans, Streptococcus group A, Streptococcus group B, Pneumococcus, Gardnerella. The medicine can maintain the activity of female reproductive tract probiotics within 24 hours after administration. The female reproductive tract probiotics comprise Lactobacillus crispatus, Lactobacillus gasseri, Lactobacillus jensenii. The medicine can improve at least one of the following clinical symptoms: vulvar itching, abnormal odor of leucorrhea, excessive leucorrhea, dyspareunia, lower abdominal pain, frequent urination, urgency, pain, anal distension, abnormal vaginal discharge, vaginal inflammation, vaginal flora imbalance.

[0086] Those skilled in the art can understand that the steps, measures and schemes in various operations, methods and processes discussed in the present application can be alternated, changed, combined or deleted. Further, other steps, measures and schemes in various operations, methods and processes discussed in the present application can also be alternated, changed, rearranged, decomposed, combined or deleted. Further, steps, measures and schemes in various operations, methods and processes in the related art can also be alternated, changed, rearranged, decomposed, combined or deleted.

[0087] The terms "first", "second" are only for the purpose of description, and cannot be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features. Therefore, the features defined with "first", "second" can explicitly or implicitly include one or more of the features. In the description of the present application, unless otherwise specified, the meaning of "a plurality of" is two or more.

[0088] The above only describes some embodiments of the present application, and it should be pointed out that, for those skilled in the art, other similar implementation manners based on the technical idea of the present application can be adopted without departing from the technical concept of the present application, and such implementation manners also belong to the protection scope of the embodiments of the present application.

Claims

1. Use of a traditional Chinese medicine composition in the preparation of a medicament for treating inflammation of the female genital tract, the traditional Chinese medicine composition comprising 50 parts of Amoracia, 50 parts of Cnidium, 30 parts of Sophora, 20 parts of Uncaria, 10 parts of Potassium Sulphate, and 8 parts of Coptis.

2. The use of claim 1, wherein the medicament comprises an extract of the traditional Chinese medicine composition, the extract being prepared by the steps of: formulating the traditional Chinese medicine composition; boiling the composition in water in a ratio of 10-12 parts of water to one part of the composition, to obtain a first filtrate; repeating the boiling of the residue to obtain a second filtrate; and combining the first and second filtrates to obtain the extract.

3. The use of claim 2, wherein the extract is prepared into a decoction, a concentrated solution, a gel or a dry powder.

4. The use of claim 2, wherein the medicament is for external use.

5. The use of claim 4, wherein the medicament is used by sitz bath, external washing or vaginal irrigation with the extract.

6. The use of claim 5, wherein the medicament is used once a day for several days.

7. The use of claim 5, wherein the medicament is capable of inhibiting the growth of at least one of the following microorganisms in the female genital tract within 20 minutes of administration: Micrococcus flavus, Prevotella, Atopobium, Escherichia coli, Staphylococcus aureus, Candida albicans, Streptococcus group A, Streptococcus group B, Pneumococcus, and Gardnerella.

8. The use of claim 5, wherein the medicament is capable of maintaining the activity of probiotics in the female genital tract within 24 hours of administration.

9. The use of claim 8, wherein the probiotics in the female genital tract comprise Lactobacillus crispatus, Lactobacillus gasseri, and Lactobacillus jensenii.

10. The use of claim 1, wherein the medicament is capable of improving at least one of the following clinical symptoms: pruritus of the vulva, abnormal odor of the leucorrhea, excessive leucorrhea, dyspareunia, lower abdominal pain, frequent urination, urgency, pain, anal distension, abnormal vaginal discharge, inflammation of the vagina, and vaginal flora imbalance. ​ ​ ​ ​

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