CN112206259B - Traditional Chinese medicine composition for treating vaginitis, traditional Chinese medicine lotion and application - Google Patents

Traditional Chinese medicine composition for treating vaginitis, traditional Chinese medicine lotion and application Download PDF

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CN112206259B
CN112206259B CN202011196040.XA CN202011196040A CN112206259B CN 112206259 B CN112206259 B CN 112206259B CN 202011196040 A CN202011196040 A CN 202011196040A CN 112206259 B CN112206259 B CN 112206259B
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梁引库
徐仲阳
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Shaanxi University of Technology
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Abstract

The invention belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition for treating vaginitis, a traditional Chinese medicine lotion and application. The invention provides a traditional Chinese medicine composition for treating vaginitis, which comprises a dandelion ethyl acetate extract, a gallnut aqueous extract, a sophora flavescens ethyl acetate extract and a coptis chinensis ethanol extract. The traditional Chinese medicine composition provided by the invention can effectively achieve the technical effect of in vitro bacteriostasis. The traditional Chinese medicine composition provided by the invention has the technical effects of inhibiting gardnerella, staphylococcus aureus and candida albicans, and also has the effects of relieving itching and diminishing swelling, and has very high safety, so that adverse reactions of patients after use can be effectively avoided; and the antipruritic effect is obviously better than that of dexamethasone.

Description

Traditional Chinese medicine composition for treating vaginitis, traditional Chinese medicine lotion and application
Technical Field
The invention belongs to the field of traditional Chinese medicines, and particularly relates to a traditional Chinese medicine composition for treating vaginitis, a traditional Chinese medicine lotion and application.
Background
Vaginitis is a disease caused by mixed pathogenic bacteria infection, and is mainly caused by staphylococcus aureus, candida albicans, gardnerella and trichomonas. Because pathogenic pathogens are more and mixed easily, the treatment difficulty of the vaginitis is higher and the vaginitis is easy to relapse. Clinically, antibiotics are used for treatment, but the use of antibiotics often causes the occurrence of antibiotic drug abuse. In addition, the irregular use of antibiotics can easily cause the disturbance of the internal environment of the vagina, and further cause the repeated attacks after the healing and the occurrence of drug resistance and the like.
Disclosure of Invention
In order to solve the problems, the invention provides a traditional Chinese medicine composition for treating vaginitis, a traditional Chinese medicine lotion and application. The traditional Chinese medicine composition provided by the invention has the outstanding technical effects of inhibiting bacteria, relieving itching and diminishing swelling, can effectively play a role in treating vaginitis, and has the characteristics of simple composition and low production cost.
In order to achieve the purpose, the invention provides the following technical scheme:
the invention provides a traditional Chinese medicine composition for treating vaginitis, which comprises the following components in parts by mass: 5-15 parts of dandelion ethyl acetate extract, 25-40 parts of gallnut aqueous extract, 20-45 parts of sophora flavescens ethyl acetate extract and 10-30 parts of coptis chinensis ethanol extract.
Preferably, the traditional Chinese medicine composition comprises the following components in parts by mass: 10 parts of dandelion ethyl acetate extract, 35 parts of Chinese gall water extract, 40 parts of sophora flavescens ethyl acetate extract and 15 parts of coptis chinensis alcohol extract.
Preferably, the traditional Chinese medicine composition comprises the following components in parts by mass: 15 parts of dandelion ethyl acetate extract, 30 parts of Chinese gall water extract, 30 parts of sophora flavescens ethyl acetate extract and 25 parts of coptis chinensis alcohol extract.
Preferably, the preparation method of the dandelion ethyl acetate extract comprises the following steps: extracting the dandelion dry powder by using ethyl acetate; the volume ratio of the dandelion dry powder to the ethyl acetate is 1 (15-20).
Preferably, the preparation method of the gallnut aqueous extract comprises the following steps: extracting the gallnut dry powder by adopting water; the volume ratio of the gallnut dry powder to water is 1 (15-20).
Preferably, the preparation method of the ethyl acetate sophora flavescens extract comprises the following steps: extracting the sophora flavescens dry powder by using ethyl acetate; the volume ratio of the sophora flavescens dry powder to the ethyl acetate is 1 (15-20).
Preferably, the preparation method of the coptis chinensis alcohol extract comprises the following steps: extracting the dry coptis powder by adopting absolute ethyl alcohol; the volume ratio of the coptis chinensis dry powder to the absolute ethyl alcohol is 1 (15-20).
Preferably, the dosage form of the traditional Chinese medicine composition comprises traditional Chinese medicine lotion.
The invention also provides a traditional Chinese medicine lotion based on the traditional Chinese medicine composition, and the traditional Chinese medicine lotion comprises the traditional Chinese medicine composition, dimethyl sulfoxide, benzoic acid, paraben and sterile water; the traditional Chinese medicine lotion comprises, by mass, 10-30 mg/mL of a traditional Chinese medicine composition, 6-9% of dimethyl sulfoxide, 0.20-0.35% of benzoic acid, 0.015-0.025% of nipagin ester and the balance of sterile water.
The invention provides application of the traditional Chinese medicine composition or the traditional Chinese medicine lotion in preparing a medicine for treating vaginitis.
The invention provides a traditional Chinese medicine composition for treating vaginitis, which comprises a dandelion ethyl acetate extract, a gallnut aqueous extract, a sophora flavescens ethyl acetate extract and a coptis chinensis ethanol extract. The traditional Chinese medicine composition provided by the invention can effectively achieve the technical effect of in vitro bacteriostasis. The application examples show that the traditional Chinese medicine composition provided by the invention has the technical effects of inhibiting gardnerella, staphylococcus aureus and candida albicans, and also has the effects of relieving itching and diminishing swelling, and the traditional Chinese medicine composition provided by the invention has very high safety and can effectively avoid adverse reactions of patients after use. Test results show that the itching relieving effect of the traditional Chinese medicine composition provided by the invention is obviously superior to that of dexamethasone.
Drawings
FIG. 1 is a front view of the rabbit vaginal mucosa irritation test;
FIG. 2 is a side view of the rabbit vaginal mucosa irritation test.
Detailed Description
The invention provides a traditional Chinese medicine composition for treating vaginitis, which comprises the following components in parts by mass: 5-15 parts of dandelion ethyl acetate extract, 25-40 parts of gallnut aqueous extract, 20-45 parts of sophora flavescens ethyl acetate extract and 10-30 parts of coptis chinensis ethanol extract. The traditional Chinese medicine composition provided by the invention has the effects of resisting inflammation, reducing swelling and relieving itching, has no adverse reaction, and can be safely used by patients.
The traditional Chinese medicine composition provided by the invention preferably comprises 5-15 parts by mass of dandelion ethyl acetate extract, and more preferably 10 parts or 15 parts by mass of dandelion ethyl acetate extract. In the present invention, the method for preparing the dandelion ethyl acetate extract preferably comprises: extracting the dandelion dry powder by using ethyl acetate; the volume ratio of the dandelion dry powder to the ethyl acetate is 1 (15-20); the preparation method preferably specifically comprises the following steps: drying the dandelion dry powder to constant weight in an environment of 70-80 ℃, crushing the dried dandelion, sieving by a 60-mesh sieve to obtain dandelion dry powder, mixing the dandelion dry powder with ethyl acetate according to the volume ratio of 1 (15-20), and soaking for 20 min. Then carrying out ultrasonic reflux extraction for 50-75 min at 50-70 ℃, preferably 60 ℃ and 60W, preferably 60min, filtering and collecting first filtrate and filter residue; performing ultrasonic reflux extraction on the filter residue for 50-75 min at 50-70 ℃, preferably at 60 ℃ and 60W, filtering and collecting second filtrate, and combining the first filtrate and the second filtrate to obtain combined filtrate; centrifuging the combined filtrate for 10min at 8000r/min, collecting supernatant, and drying the supernatant to obtain herba Taraxaci ethyl acetate extract; the relative density of the dandelion ethyl acetate extract at 37 ℃ is preferably 1.10-1.30 g/ml; the drying operation comprises vacuum drying at 70 ℃.
The traditional Chinese medicine composition provided by the invention comprises, by mass, preferably 25-40 parts of a gallnut aqueous extract, and more preferably 30 parts or 35 parts. In the present invention, the preparation method of the aqueous extract of gallnut preferably comprises: extracting the gallnut dry powder by adopting water; the volume ratio of the gallnut dry powder to water is 1 (15-20); the preparation method particularly preferably comprises the following steps: drying the gallnut to constant weight in an environment of 70-100 ℃, crushing the gallnut dried to constant weight, sieving with a 60-mesh sieve to obtain dry gallnut powder, mixing the dry gallnut powder with water according to a volume ratio of 1 (15-20), and further preferably 1:20, and extracting to obtain the water extract of the gallnut.
The method for extracting the mixed solution preferably comprises the following steps: and carrying out reflux extraction on the mixed solution at 85-100 ℃, preferably 100 ℃, for 70-90 min, preferably 90min, filtering and collecting the first filtrate and filter residue. And carrying out reflux extraction on the filter residue for 70-90 min, preferably 90min, at the temperature of 85-100 ℃ and preferably 100 ℃, filtering and collecting a second filtrate. Mixing the first filtrate and the second filtrate to obtain a mixed filtrate, centrifuging the mixed filtrate for 10-15 min under the condition of 7000-8000 r/min, taking supernatant, and drying the supernatant to obtain a Chinese gall water extract; the relative density of the water extract of the gallnut is preferably 1.12-1.32 g/ml at 37 ℃; the drying operation comprises vacuum drying at 70 ℃.
The traditional Chinese medicine composition provided by the invention preferably comprises 20-45 parts by mass of the ethyl acetate sophora flavescens extract, and more preferably 30 parts or 40 parts by mass of the ethyl acetate sophora flavescens extract. In the present invention, the preparation method of the ethyl acetate extract of sophora flavescens preferably comprises: extracting the sophora flavescens dry powder by using ethyl acetate; the volume ratio of the sophora flavescens dry powder to the ethyl acetate is 1 (15-20); the preparation method particularly preferably comprises the following steps: drying whole plant of radix Sophorae Flavescentis at 70 deg.C to constant weight, pulverizing dried radix Sophorae Flavescentis to constant weight, and sieving with 60 mesh sieve to obtain radix Sophorae Flavescentis dry powder. Mixing the sophora flavescens dry powder with water according to the volume ratio of 1 (15-20), and preferably 1:20, soaking for 20min, and extracting.
The extraction method preferably comprises: and (3) performing ultrasonic reflux extraction on the soak solution at 50-70 ℃ and 60W, preferably 60 ℃ for 50-75 min, preferably 60min, and filtering to collect first filtrate and filter residue. And performing ultrasonic reflux extraction on the filter residue at 50-70 ℃ and 60W, preferably 60 ℃ for 50-75 min, preferably 60min, filtering and collecting a second filtrate. Combining the first filtrate and the second filtrate to obtain a combined filtrate, centrifuging the combined filtrate for 10-15 min under the condition of 7000-8000 r/min, taking supernatant, and drying the supernatant to obtain a sophora flavescens ethyl acetate extract; the relative density of the lightyellow sophora root ethyl acetate extract at 37 ℃ is preferably 1.15-1.35 g/ml; the drying operation comprises vacuum drying at 70 ℃.
The traditional Chinese medicine composition provided by the invention preferably comprises 10-30 parts by mass of coptis chinensis ethanol extract, and more preferably 15 or 25 parts by mass of coptis chinensis ethanol extract. In the present invention, the preparation method of the golden thread alcohol extract preferably comprises: extracting the dry coptis powder by adopting absolute ethyl alcohol; the volume ratio of the coptis chinensis dry powder to the absolute ethyl alcohol is 1 (15-20); the preparation method particularly preferably comprises the following steps: drying the coptis chinensis in an environment at 70 ℃ to constant weight, crushing the coptis chinensis dried to constant weight, sieving the crushed coptis chinensis with a 60-mesh sieve to obtain coptis chinensis dry powder, mixing and soaking the coptis chinensis dry powder and absolute ethyl alcohol according to a volume ratio of 1 (15-20), and further preferably 1:20 for 20min, and extracting to obtain the coptis chinensis alcohol extract.
The method for extracting the mixed solution preferably comprises the following steps: and performing ultrasonic reflux extraction on the soak solution at 50-70 ℃ and 60W, preferably 60 ℃ for 50-75 min, preferably 60min, and filtering to collect first filtrate and filter residue. And performing ultrasonic reflux extraction on the filter residue at 50-70 ℃ and 60W, preferably 60 ℃, for 50-75 min, preferably 60min, filtering and collecting a second filtrate. And combining the first filtrate and the second filtrate to obtain a combined filtrate, centrifuging the combined filtrate for 10-15 min under the condition of 7000-8000 r/min, taking a supernatant, and drying the supernatant to obtain the coptis chinensis ethanol extract. The relative density of the coptis chinensis ethanol extract at 37 ℃ is preferably 1.16-1.36 g/ml; the drying operation comprises vacuum drying at 70 ℃.
The extraction method of the medicine extract provided by the invention has the advantages of stable process, high target product acquisition rate, good result reproducibility, good quality of the obtained product and easy realization of industrial production.
The traditional Chinese medicine composition provided by the invention can obviously improve the inhibition effect of the traditional Chinese medicine composition on staphylococcus aureus, gardnerella and candida albicans through a specific traditional Chinese medicine, a specific extraction method and a specific proportion; meanwhile, the traditional Chinese medicine composition provided by the invention has the characteristic of easily obtained raw materials; moreover, the traditional Chinese medicine composition provided by the invention is simple in composition and low in cost, but the efficacy of the traditional Chinese medicine composition is superior to that of the traditional Chinese medicine composition on the market.
In the invention, the dosage form of the traditional Chinese medicine composition preferably comprises traditional Chinese medicine lotion.
The invention also provides a traditional Chinese medicine lotion based on the traditional Chinese medicine composition, and the traditional Chinese medicine lotion comprises the traditional Chinese medicine composition, dimethyl sulfoxide, benzoic acid, paraben and sterile water; the traditional Chinese medicine lotion comprises, by mass, 10-30 mg/mL of a traditional Chinese medicine composition, 6-9% of dimethyl sulfoxide, 0.20-0.35% of benzoic acid, 0.015-0.025% of paraben and the balance of sterile water. The mass concentration of the traditional Chinese medicine composition in the traditional Chinese medicine lotion is preferably 15-25 mg/mL, and more preferably 15mg/mL or 20 mg/mL.
In the invention, when the mass concentration of the traditional Chinese medicine composition in the traditional Chinese medicine lotion is 10-30 mg/mL, the traditional Chinese medicine composition preferably comprises the following components in parts by mass:
5-15 parts of dandelion ethyl acetate extract, 25-40 parts of gallnut aqueous extract, 20-45 parts of sophora flavescens ethyl acetate extract and 10-30 parts of coptis chinensis ethanol extract;
in the invention, when the mass concentration of the traditional Chinese medicine composition in the traditional Chinese medicine lotion is 15-25 mg/mL, the traditional Chinese medicine composition preferably comprises the following components in parts by mass: 10-15 parts of dandelion ethyl acetate extract, 30-40 parts of gallnut aqueous extract, 25-45 parts of sophora flavescens ethyl acetate extract and 15-25 parts of coptis chinensis ethanol extract;
in the invention, when the mass concentration of the traditional Chinese medicine composition in the traditional Chinese medicine lotion is 15mg/mL, the traditional Chinese medicine composition preferably comprises the following components in parts by mass: 10 parts of dandelion ethyl acetate extract, 35 parts of Chinese gall water extract, 40 parts of sophora flavescens ethyl acetate extract and 15 parts of coptis chinensis ethanol extract;
in the invention, when the mass concentration of the traditional Chinese medicine composition in the traditional Chinese medicine lotion is 20mg/mL, the traditional Chinese medicine composition preferably comprises the following components in parts by mass: 15 parts of dandelion ethyl acetate extract, 30 parts of Chinese gall aqueous extract, 30 parts of sophora flavescens ethyl acetate extract and 25 parts of coptis chinensis ethanol extract.
When the mass concentration of the traditional Chinese medicine composition in the traditional Chinese medicine lotion provided by the invention is 10-30 mg/mL, the traditional Chinese medicine lotion has the best effects of detumescence, itching relieving and bacteriostasis.
The traditional Chinese medicine lotion provided by the invention has high content of effective components, can effectively achieve the technical effects of bacteriostasis, itching relieving, detumescence and the like in vitro, is safe and non-irritant, and conforms to various regulations of routine inspection of liquid preparations in 2015 edition of Chinese pharmacopoeia.
In the invention, the dandelion, the gallnut, the sophora flavescens, the coptis chinensis, the dimethyl sulfoxide, the nipagin benzoate, the sterile water and the like are purchased from markets, wherein the dandelion, the gallnut, the sophora flavescens and the coptis chinensis are all traditional Chinese medicines collected in 2015 edition of Chinese pharmacopoeia and all meet the corresponding quality requirements.
The invention provides application of the traditional Chinese medicine composition or the traditional Chinese medicine lotion in preparing a medicine for treating vaginitis. The traditional Chinese medicine composition or the traditional Chinese medicine lotion provided by the invention can effectively inhibit the growth and reproduction of pathogenic bacteria of vaginal infection caused by staphylococcus aureus, gardnerella, candida albicans and the like to achieve the technical effect of treating vaginitis, and meanwhile, the traditional Chinese medicine composition or the traditional Chinese medicine lotion provided by the invention has the effects of relieving itching, reducing swelling and the like, not only achieves the corresponding curative effect, but also greatly improves the use feeling of patients.
For further illustration of the present invention, the following detailed description of the present invention will be made with reference to the drawings and examples to describe a Chinese medicinal composition, a Chinese medicinal lotion and an application for treating vaginitis, which should not be construed as limiting the scope of the present invention.
Example 1
Preparing the dandelion ethyl acetate extract:
drying herba Taraxaci at 70 deg.C to constant weight, pulverizing, and sieving with 60 mesh sieve to obtain herba Taraxaci dry powder; placing 20g of dandelion dry powder in a beaker, adding ethyl acetate according to the volume ratio of 1:20, soaking for 20min, performing ultrasonic reflux extraction at 60w at 60 ℃ for 60min, and filtering and collecting filtrate and filter residue by using qualitative filter paper; extracting the residue with the same method under reflux for 60min, and mixing the filtrates; the combined filtrates were mixed at 8000r min -1 Centrifuging for 10min, collecting supernatant, drying at 70 deg.C under vacuum, and storing at 4 deg.C.
Preparing a gallnut aqueous extract:
drying Galla chinensis at 70 deg.C to constant weight, pulverizing, and sieving with 60 mesh sieve to obtain Galla chinensis dry powder; putting 20g of nutgall dry powder into a beaker, adding distilled water according to the volume ratio of 1:20, performing reflux extraction at 100 ℃ for 90min, filtering by using qualitative filter paper, and collecting filtrate and filter residue; the filter residue was treated in the same manner as above at a volume ratio of 1:15Extracting with distilled water for 60min, filtering with qualitative filter paper, and mixing the filtrates; mixing filtrates at 8000r min -1 Centrifuging for 10min, collecting supernatant, drying the supernatant in vacuum at 70 deg.C, and storing at 4 deg.C.
Preparing the lightyellow sophora root ethyl acetate extract:
drying radix Sophorae Flavescentis at 70 deg.C to constant weight, pulverizing, and sieving with 60 mesh sieve to obtain radix Sophorae Flavescentis dry powder; taking 20g of radix sophorae flavescentis dry powder, adding ethyl acetate according to a volume ratio of 1:20 into a beaker, soaking for 20min, performing ultrasonic reflux extraction at 60 ℃ for 60min, filtering by qualitative filter paper, and collecting filtrate and filter residue; extracting the residue with the same method under reflux for 60min, mixing the filtrates, and mixing the filtrates at 8000r min -1 Centrifuging for 10min, collecting supernatant, vacuum drying at 70 deg.C, and storing at 4 deg.C.
Preparing the ethanol extract of the coptis:
drying Coptidis rhizoma at 70 deg.C to constant weight, pulverizing, and sieving with 60 mesh sieve to obtain Coptidis rhizoma dry powder; taking 20g of coptis chinensis dry powder, adding absolute ethyl alcohol according to a volume ratio of 1:20 into a beaker, soaking for 20min, performing ultrasonic reflux extraction at 60 ℃ for 60min, filtering by qualitative filter paper, and collecting filtrate and filter residue; extracting the residue with the same method under reflux for 60min, mixing the filtrates, and mixing the filtrates at 8000r min -1 Centrifuging for 10min, collecting supernatant, vacuum drying at 70 deg.C, and storing at 4 deg.C.
Example 2
Taking 1.5 parts of dandelion ethyl acetate extract, 6 parts of Chinese gall aqueous extract, 6 parts of sophora flavescens ethyl acetate extract and 3 parts of coptis chinensis alcohol extract provided in example 1.
Dissolving herba Taraxaci ethyl acetate extract 1.5mg, Galla chinensis water extract 6mg, radix Sophorae Flavescentis ethyl acetate extract 6mg, and Coptidis rhizoma ethanol extract 3mg in 1mL sterile water containing 8% DMSO, 0.25% benzoic acid and 0.02% nipagin to obtain Chinese medicinal lotion.
Example 3
2 parts of dandelion ethyl acetate extract, 8 parts of Chinese gall water extract, 8 parts of sophora flavescens ethyl acetate extract and 4 parts of coptis chinensis alcohol extract provided in example 1 are taken.
2mg of dandelion ethyl acetate extract, 8mg of Chinese gall water extract, 8mg of sophora flavescens ethyl acetate extract and 4mg of coptis chinensis alcohol extract are dissolved in 1mL of sterile water containing 8% of DMSO, 0.25% of benzoic acid and 0.02% of nipagin, thus obtaining the traditional Chinese medicine lotion.
Example 4
2 parts of dandelion ethyl acetate extract, 8 parts of Chinese gall water extract, 8 parts of sophora flavescens ethyl acetate extract and 4 parts of coptis chinensis alcohol extract provided in example 1 are taken.
2.5mg of dandelion ethyl acetate extract, 10mg of Chinese gall water extract, 10mg of sophora flavescens ethyl acetate extract and 4mg of coptis chinensis alcohol extract are dissolved in 1mL of sterile water containing 8% of DMSO, 0.25% of benzoic acid and 0.02% of nipagin, thus obtaining the traditional Chinese medicine lotion.
Application example 1
In vitro bacteriostasis:
test medication: extracts were prepared in the manner described in example 1, and prepared at the mass ratio of the extracts described in example 2, to prepare a lotion stock solution having a concentration of 256 mg/mL.
The test process comprises the following steps: inoculating staphylococcus aureus to an LB liquid culture medium; inoculating Candida albicans to a Sabouraud's liquid culture medium; gardnerella are inoculated to HTM liquid medium.
The test drugs were added to 10mL test tubes containing 2mL of the corresponding liquid medium at final concentrations of 128mg/mL, 64mg/mL, 32mg/mL, 16mg/mL, 8mg/mL, 4mg/mL, 2mg/mL, 1mg/mL, 0.5mg/mL, 0.25mg/mL, and 0mg/mL, respectively. Inoculating the bacterial suspension at 5% (V/V), and culturing at 37 deg.C for 200r min -1 Culturing for 24h under the conditions of (1). The drug concentration in the control group was 0 mg/mL. Compared with a control, the lowest drug concentration for completely inhibiting the bacterial growth is taken as MIC, liquid in a corresponding tube is fully dipped by using an aseptic inoculating loop due to the drug color influence and crude extraction turbidity influence, the liquid is spread on a corresponding agar culture medium, the liquid is cultured overnight at 37 ℃ for 12h, no bacteria grow at the streak line, the MIC of the drug is determined to be extracted, and the detection result is shown in Table 1.
TABLE 1 MIC of drug extract against pathogenic bacteria as provided in example 4 of example 1
Figure BDA0002754037590000091
As can be seen from Table 1, the Chinese medicinal composition provided by the invention can effectively inhibit the growth of Gardnerella, Staphylococcus aureus and Candida albicans.
Application example 2
The itching relieving effect is as follows:
test medication: extracts were prepared in the manner as in example 1, and formulated at the mass ratio of the extracts in example 2 to prepare the drugs in the high dose group at a concentration of 26.0mg/mL, the medium dose group at 13.0mg/mL and the low dose group at 6.5mg/mL, respectively.
Positive control: dexamethasone was dosed per 100mg/100g body weight.
40 guinea pigs are taken as test objects, and are randomly divided into a model control group, a dexamethasone positive drug group and test drug high, medium and low dose groups according to sex and weight, wherein each group comprises 8 guinea pigs and is half male and female. The right hind instep of each guinea pig was shaved 24h before molding (about 1 cm) 2 ) In the experiment, the hair shaving positions of all groups are pricked by a triangular needle on the day with mild blood leakage as a degree, and the dexamethasone positive medicament and the low, medium and high dosage medicaments of the traditional Chinese medicine formula are respectively smeared on the affected part with the dosage of 0.1g/100g of body weight for 2 times of continuous administration with the interval time of 1h each time. After 15min of the last administration, 0.01% histamine phosphate is smeared at the stabs, and then 0.02%, 0.03%, 0.04%, 0.05%, 0.1%, 0.2% and 0.3% histamine phosphate are smeared at intervals of 3min in sequence until the guinea pig licks the right hind foot, if the guinea pig licks the hind foot to 0.3%, the smearing of the histamine phosphate is stopped when no licking is found, the total amount (mg) of the histamine phosphate is recorded as the scratchiness threshold, and the scratching times in unit time are recorded for carrying out biological statistics and data analysis. The statistical results are shown in tables 2 and 3 (note: 3min is one unit time).
TABLE 2 analysis of scratchiness threshold/mg of drug (in total X. + -. S)
Figure BDA0002754037590000101
The control group has p <0.05, indicated by "-"; when p <0.01, it is denoted by "×".
Comparing the positive control group with the traditional Chinese medicine group, wherein p is less than 0.05 and is represented by "#"; when p is less than 0.01, it is indicated by "##".
TABLE 3 scratchiness threshold comparison/mg (used in total)
Figure BDA0002754037590000102
Is shown)
Figure BDA0002754037590000103
When p is less than 0.05, the control group is marked with an asterisk when compared with the traditional Chinese medicine group; when p <0.01, indicated by "×".
When the p is less than 0.05, the positive control group is represented by "#" when being compared with the traditional Chinese medicine group; when p <0.01, it is denoted by "##".
The results of scratching times per unit time in Table 2 show that the low-dose group of the Chinese medicinal composition has the best antipruritic effect and is superior to the dexamethasone group. The high and medium dose groups of the test drug also had antipruritic effects, but no significant difference compared to dexamethasone (p > 0.05). The results in Table 3 show that the antipruritic effect of the high and low dosage groups of the medicine is obvious, and the antipruritic effect of the medium dosage group of the medicine is extremely obvious. In conclusion, the two indexes of scratching frequency per unit time and histamine phosphate total amount of the guinea pigs are examined to determine that the itching relieving effect of the traditional Chinese medicine low-dose group is the best and is better than that of a dexamethasone positive control group. But the high, medium and low concentrations all have good antipruritic effect.
Application example 3
Detumescence effect:
test medication: preparing extracts in the manner of example 1, configuring the mass ratio of the extracts in example 2, and preparing high-dose group medicaments with the concentration of 26.0mg/mL respectively; 13.0mg/mL of the medium dose group drug; 6.5mg/mL of the low dose group drug.
Positive control: dexamethasone was dosed per 100mg/100g body weight.
Rats were randomized into five groups using rats as experimental subjects: model control group, dexamethasone positive control group, and high, medium, and low dosage groups of Chinese medicinal composition, wherein each group contains 8 animals. 3% formaldehyde solution is prepared as a inflammatory agent, and 100 mu L of the inflammatory agent is absorbed by a 1 mL-specification syringe and is respectively injected into the toes of a rat to establish a swelling model. Before molding, the thickness (mm) of the toes of each group of rats was measured with a vernier caliper, respectively. Then, the rats of the model control group, the dexamethasone positive control group and the high, medium and low dosage groups of the traditional Chinese medicine are respectively injected with an inflammatory agent to cause inflammatory reaction, the toe thickness after inflammation of the rats of the model control group is respectively measured at 0.5h, 1h, 2h, 4h and 6h, and the time of the peak period (the highest swelling degree) of the inflammatory agent is obtained. At peak time, the dexamethasone positive control group is smeared with 0.25mg of the drug; the rats with high, medium and low doses of traditional Chinese medicine are respectively and uniformly smeared on the affected part of inflammation. After the medicine is smeared, the toe thickness of the rats is measured by observing the swelling fading condition within short time of 0.5h, 1h, 2h, 4h and 6h respectively. The following day, three consecutive days, once in the morning, the swelling thickness of the toes of each group of rats was measured at fixed time in the afternoon, and the recovery time was counted, and the statistical results are shown in table 4. The time to resolution of swelling was analyzed for the administration group and the model group and expressed as the swelling rate.
Note: the swelling rate is (thickness of postinflammatory metatarsal-thickness of proinflammatory metatarsal)/thickness of proinflammatory metatarsal × 100%.
TABLE 4 Effect of drugs on rat plantar swelling (for P evaluation)
Figure BDA0002754037590000111
Is shown)
Figure BDA0002754037590000112
Figure BDA0002754037590000121
When the model control group is compared with the traditional Chinese medicine group, when p is less than 0.05, the expression is marked by a letter'; when p <0.01, it is denoted by "×".
When the p is less than 0.05, the positive control group is represented by "#" when being compared with the traditional Chinese medicine group; when p <0.01, it is denoted by "##".
From the above, the high dose and the medium dose of the medicine have obvious detumescence effect, and have no obvious difference compared with the dexamethasone positive control.
Application example 4
Safety:
experimental rabbits (new zealand white rabbits) were randomly divided into two groups: the infected group and the control group were 3 per group. Before the experiment, the vaginal orifice of the animal is checked for secretion, congestion, edema and other injury conditions. If inflammation and/or injury occur, it is discarded. The experimental animal can be tested after being normal and without problems.
The drugs used in the drug exposure group were prepared into liquid medicine with the concentration of 52mg/mL for standby based on the mass ratio of the drug extracts in example 4, and the control group used sterile normal saline.
An 8cm blunt hose was connected to a syringe, and 2mL of the solution was aspirated by the syringe to fill the catheter and the syringe for future use. The animal is fixed in a prone position, the perineum and the vaginal orifice are exposed, the catheter is wetted and then gently rubbed and inserted into the vagina (4-5 cm), 2mL of liquid medicine is slowly injected by using an injector, and the catheter is drawn out to finish the contamination. Control animals were treated with saline as well. The infection is repeated every 24h for 5 days continuously. In the case of injection, sometimes the test solution may overflow due to animal differences and may be wiped off with a sterilized cotton. 24h after the last infection. The animals were sacrificed by air embolism, the whole vagina was removed by laparotomy, and the animals were incised longitudinally and visually examined for signs of congestion, edema, etc. Then the vaginal tissue is put into 10% formalin solution for fixation for more than 24H, tissue sections on the front face and the side face of the vagina are selected, and after H & E staining (staining pictures are shown in figures 1 and 2), histopathological examination is carried out. Statistical analysis was performed using the vaginal mucosal irritation response scoring criteria (table 5) and the vaginal mucosal irritation intensity grading table (table 6), and the average scores of the toxicant-infected group and the control group are shown in table 7.
TABLE 5 vaginal mucosal irritation response Scoring criteria
Figure BDA0002754037590000131
Figure BDA0002754037590000141
TABLE 6 vaginal mucosal irritation Strength grading
Figure BDA0002754037590000142
TABLE 7 vaginal mucosa score of New Zealand white rabbits injected with drug
Figure BDA0002754037590000143
According to the evaluation of 'vagina mucosa stimulation intensity grading' in 'disinfection technical specification' (2019 edition), the stimulation index of the medicine provided by the invention to the vagina mucosa of the New Zealand white rabbit is 0.2, which indicates that the medicine provided by the invention has no stimulation to the vagina mucosa. Observation under an optical microscope of pathological tissue sections shows that the vaginal mucosa surface of the experimental rabbit is coated with squamous epithelium, the hierarchical structure is clear, the white blood cells of the inherent membrane are slightly increased, the red blood cells are more frequent, and the drug administration and the control sections have no obvious difference. It shows that the medicine has no irritation to vaginal tissue.
In conclusion, the traditional Chinese medicine composition and the traditional Chinese medicine lotion provided by the invention can obviously inhibit vaginal inflammation caused by candida albicans, gardnerella and staphylococcus aureus, and have good itching relieving, swelling diminishing and anti-inflammatory effects without stimulation.
Although the present invention has been described with reference to the preferred embodiments, it should be understood that various changes and modifications can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims.

Claims (6)

1. A traditional Chinese medicine composition for external use for treating vaginitis caused by Gardner bacteria, Staphylococcus aureus and Candida albicans is prepared from the following components in parts by mass: 5-15 parts of dandelion ethyl acetate extract, 25-40 parts of gallnut aqueous extract, 20-45 parts of sophora flavescens ethyl acetate extract and 10-30 parts of coptis chinensis ethanol extract;
the preparation method of the dandelion ethyl acetate extract comprises the following steps: extracting the dandelion dry powder by using ethyl acetate; the volume ratio of the dandelion dry powder to the ethyl acetate is 1 (15-20);
the preparation method of the gallnut aqueous extract comprises the following steps: extracting the gallnut dry powder by adopting water; the volume ratio of the gallnut dry powder to water is 1 (15-20);
the preparation method of the lightyellow sophora root ethyl acetate extract comprises the following steps: extracting the sophora flavescens dry powder by using ethyl acetate; the volume ratio of the sophora flavescens dry powder to the ethyl acetate is 1 (15-20);
the preparation method of the coptis chinensis ethanol extract comprises the following steps: extracting the dry coptis powder by adopting absolute ethyl alcohol; the volume ratio of the coptis chinensis dry powder to the absolute ethyl alcohol is 1 (15-20).
2. The external traditional Chinese medicine composition as claimed in claim 1, which is prepared from the following components in parts by mass: 10 parts of dandelion ethyl acetate extract, 35 parts of Chinese gall water extract, 40 parts of sophora flavescens ethyl acetate extract and 15 parts of coptis chinensis alcohol extract.
3. The external traditional Chinese medicine composition as claimed in claim 1, which is prepared from the following components in parts by mass: 15 parts of dandelion ethyl acetate extract, 30 parts of Chinese gall water extract, 30 parts of sophora flavescens ethyl acetate extract and 25 parts of coptis chinensis alcohol extract.
4. The topical Chinese medicinal composition according to any one of claims 1 to 3, wherein the dosage form of the topical Chinese medicinal composition comprises a Chinese medicinal lotion.
5. A traditional Chinese medicine lotion based on the traditional Chinese medicine composition for external use of any one of claims 1 to 4, wherein the components of the traditional Chinese medicine lotion comprise the traditional Chinese medicine composition for external use of any one of claims 1 to 4, dimethyl sulfoxide, benzoic acid, nipagin ester and sterile water; the mass concentration of the external traditional Chinese medicine composition in the traditional Chinese medicine lotion is 10-30 mg/mL, the mass percentage of dimethyl sulfoxide is 6-9%, the mass percentage of benzoic acid is 0.20-0.35%, the mass percentage of nipagin ester is 0.015-0.025%, and the balance is sterile water.
6. Use of the topical Chinese medicinal composition of any one of claims 1 to 4 or the Chinese medicinal lotion of claim 5 for the preparation of a medicament for the treatment of vaginitis caused by gardnerella, staphylococcus aureus and candida albicans.
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
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