CN101028288A - Health-care mask - Google Patents

Abstract

A Chinese medicine in the form of capsule or effervescent tablet for regulating bacterial flora and preventing or treating bacterial vaginosis is prepared from lactobacillus delbrueckii (5-15 wt. portions), lactobacillus acidophilus (5-15), lactose (500-. Has the effects of regulating vaginal flora imbalance, and effectively inhibiting external attack bacteria and a few of excessively-propagated vaginal flora.

Description

Jianmianjing

Technical field: The present invention relates to a probiotic preparation for adjusting flora to prevent or treat bacterial vaginosis, which is used to improve vaginal micro-ecological balance and enhance immunity, and relates to a process for preparing the preparation.

Background technology: Bacterial vaginosis (BV), nonspecific vaginitis (NSV) or dysbacteriosis vaginitis (DV) is a common and frequently-occurring disease in women. It is usually treated with antibiotics in clinic, but the effect is not ideal. The application of antibiotics itself can also cause flora imbalance, causing the disease to persist and bring great suffering to patients. Other gynecological infectious diseases such as various sexually transmitted diseases, trichomonal vaginitis, and fungal vaginitis can also lead to vaginal flora imbalance after being treated with a large amount of antibiotics, causing flora imbalance vaginitis, and antibiotics are often used to treat them. . The existing research on probiotics is also limited to the research on a single strain, and there is no mutual promotion and symbiosis of the probiotics, and there is no coordination to promote the adjustment of probiotics to prevent or treat bacterial vaginosis. A probiotic preparation and process for preventing or treating bacterial vaginosis.

Invention content:

From the perspective of microecology, adjusting the therapy with ecological preparations can help correct and protect the composition and proportion of normal flora in the vagina, restore its natural ability to resist foreign bacteria intrusion, and promote its own self-purification effect to treat such diseases an inevitable trend. The invention is a probiotic preparation for vaginal health care or prevention and treatment of bacterial vaginosis by using beneficial bacteria group and promoting the growth and effectiveness of other beneficial bacteria. Species: Lactobacillus debrueckii subsp. lactis, Lactobacillus acidophilus. Function: It has the function of regulating the imbalance of vaginal flora, and inhibits the minority populations in the vaginal flora that invades bacteria and overgrows. Prevent or treat bacterial vaginosis. The probiotic preparation for vaginal health care or prevention and treatment of bacterial vaginosis of the present invention contains 5-15 parts of Lactobacillus delbrueckii, 5-15 parts of Lactobacillus acidophilus, 500-700 parts of lactose, stachys 100-300 parts of sugar, fully mixed, containing 500-1500 million/g probiotic viable bacteria, divided into capsules and compressed tablets.

The invention relates to a microbiological experiment, a microecological experiment and a production process research experiment on two bacterial strains. Microbiological experiments include strain isolation, identification, biological characteristics, metabolites, drug resistance, metabolites, H ₂ O ₂ production and plasmid detection, etc. Ecological effect tests include normal value investigation, adhesion, colonization, biological antagonism, model test, safety test, stability test and stimulation test, etc. Production process research includes research on seeds, fermentation freeze-drying and formulation product stability and process stability. Basic research also includes the study of the mechanism of action.

This study conducted a relatively comprehensive and systematic study on the micro-community and micro-ecosystem of healthy vagina. The results of the study proved that Lactobacillus is the dominant micropopulation of the vagina, and other micropopulations such as Enterobacteriaceae, Staphylococcus, Streptococcus, yeast and mycoplasma are disadvantageous populations. Lactobacilli in the vagina gradually increase during the first, second and third trimesters of pregnancy. Bacterial vaginitis (BV), non-specific vaginitis (NSV) or dysbacteriosis vaginitis (DV) all have a certain phenomenon of vaginal flora imbalance, which is mainly manifested by the decrease or disappearance of Lactobacillus and the increase or proportion of other species. out of tune. Lactobacillus vaginalis is one of the important physiological indicators during pregnancy and a means of defense against infection.

The two strains involved in the present invention, Lactobacillus acidophilus and Lactobacillus delbrueckii, have been studied to show that both strains can exert their action mechanism through adhesion space occupation, adhesion inhibition, production of lactic acid and H ₂ O ₂ . Through the study of pharmacokinetics, pharmacodynamics, safety and production technology, the two strains are non-toxic, harmless, safe and effective, and the microecological preparations made of the two strains can be applied to the treatment of bacterial vaginosis. treat.

Clinical studies have proved that Lactobacillus acidophilus and Lactobacillus delbrueckii preparations are better than or no less effective than metronidazole (control group) in treating bacterial vaginosis and trichomonas vaginitis, and the recurrence rate is low, and there is no antibiotic resistance. Drug properties, causing flora imbalance and toxic reactions and other side effects. Among them, the total effective rate for bacterial vaginosis is 92.0%; while metronidazole is 83.7%, and the total effective rate for trichomonas vaginitis is 96.7%.

The probiotics manufacturing process of the present invention. 5-15 parts of Lactobacillus delbrueckii, 5-15 parts of Lactobacillus acidophilus, 500-700 parts of lactose, 100-300 parts of stachyose, fully mixed with 5-1.5 billion live probiotics/g, divided Capsules, compressed tablets. The preparation process of single bacterial powder is as follows:

(1) Preparation of Lactobacillus delbrueckii bacteria powder: Insert the seed solution of Lactobacillus delbrueckii strain into sterilized 1%-3% glucose by weight, 0.5-1.5% peptone, 0.5-1% yeast extract, 0.2-0.5% sodium acetate, 0.2-0.5% ammonium citrate medium, after anaerobic fermentation at 36°C-39°C, centrifuge to separate the bacteria, which will account for 25-25% of the weight of the protection solution Add 40% of the bacterium into the freeze-drying protective solution containing 3-5% of sodium glutamate and 5-15% of fat milk powder by weight, mix and freeze-dry to obtain the Lactobacillus delbrueckii bacteria powder.

(2) Preparation of Lactobacillus acidophilus bacterium powder: Insert the seed solution of Lactobacillus acidophilus strain containing 1%-3% glucose by weight, 0.5-1.5% peptone, 0.5-1% yeast by weight 1-5% inoculum After anaerobic fermentation at 32°C-36°C in the extract medium, the bacteria are separated by centrifugation, and the bacteria accounting for 30-45% of the weight of the protection solution are added with 100-200mM trehalose, acid neutralizing substances and antioxidant substances, etc. in the freeze-drying protection solution, mix, stand for 2-4 hours, and freeze-dry to obtain Lactobacillus acidophilus bacteria powder.

Due to the different types of lactobacilli carried in the vagina of different groups of people, the probability of colonization in the vagina is small when a single lactobacillus is used. The present invention adopts two kinds of lactobacilli that have been proved to have the effect of regulating vaginal flora imbalance as the production strains of this preparation , can increase the probability of Lactobacillus colonization in different patients with bacterial vaginosis. Greatly improve the curative effect. At the same time, the added lactose and trehalose can improve and promote the survival rate of bacteria and rapid reproduction in vagina.

The relevant research of the present invention is as follows:

1. Qualitative and quantitative research on vaginal Lactobacillus in healthy women (Table 1)

Table 1 The isolation rate and average number of viable bacteria (log10n/ml) of various lactobacilli type Number of plants Rate(%) X±sD Lactobacillus acidophilus Lactobacillus salivarius Lactobacillus fructose Lactobacillus delbrueckii Lactobacillus reisei Lactobacillus casei Lactobacillus cellobiosus 196554432 33.310.58.88.87.07.05.33.5 7.68±1.416.49±1.428.06±1.716.55±0.798.11±0.637.3349±0.90525.5658±1.82295.0947±0.5976

Note: In this experiment, a total of 57 samples of non-pregnant and childbearing women aged 20-25 years old and without vaginal diseases were collected, of which 43 samples were isolated from Lactobacillus, and the isolation rate was 75.44%.

3. Adhesion of bacteria to vaginal epithelial cells under different pH conditions. Several common normal flora and Lactobacillus isolated from the vagina were tested for their adhesion to vaginal epithelial cells under different pH conditions. The results are shown in Table 2. At pH3 .5, Lactobacillus has the strongest adhesion to vaginal epithelial cells, Staphylococcus, Enterobacter, and Candida have the strongest adhesion to vaginal epithelial cells at pH 7.2.

Table 2 Adhesion ability of different bacteria to vaginal epithelium (x±s) pH3.5 pH4.5 pH5.5 pH7.2 Lactobacillus Staphylococcus Enterobacter Candida 77.89±7.91 ^* 8.35±1.94 ^* 6.17±1.86 ^* 5.03±2.73 ^* 59.33±6.2914.77±4.6111.73±2.648.6±3.24 45.79±8.3623.94±5.3117.83±1.837.64±3.42 40.19±12.9831.4±7.6726.71±1.9415.89±2.52

*: P<0.05 compared with pH7.2.

With the increase of pH value, the adhesion ability of Lactobacillus to epithelial cells weakened, while the adhesion ability of Staphylococcus, Enterobacter and Candida to epithelial cells increased.

3. Study on the therapeutic mechanism of Lactobacillus preparations on non-specific vaginitis The results of using this preparation to treat non-specific vaginitis showed that the pH and cleanliness of vaginitis before treatment were significantly higher than those of the control group and the treatment group, and the pH of the vagina after treatment was significantly lower than that of the control group. For the control group, see Table 3.

Table 2 Comparison of vaginal pH, cleanliness, and percentage of G ⁺ bacilli control group Vaginitis group P value Before treatment After treatment Control group and before treatment Control group and after treatment Before and after treatment average pH 4.00±0.27 4.95±0.32 3.87±0.18 <0.001 <0.02 <0.001 average cleanliness 1.28±0.46 2.0±0.50 1.18±0.40 <0.001 >0.05 <0.001

Percentage of G ⁺ bacilli 97.01±4.48 72.91±11.22 98.24±4.86 ＜0.001 ＞0.05 ＜0.001

Note: Normal control group: 32 cases, aged 24-38 years, excreting gynecological diseases, no recent history of antibiotic application; treatment group: 32 cases, aged 28-41 years, clinically diagnosed with non-specific vaginitis.

4. Comparison of the average logarithmic values of positive vaginal microorganisms in each group before and after treatment with this preparation. After treatment, vaginal Lactobacillus was significantly higher than that of the control group, and Staphylococcus was significantly lower than that of the control group. see table 4

Table 4 Comparison of positive average logarithmic values of each bacteria (X±SD) Bacteria name control group Before treatment After treatment P value Control and before treatment Control and after treatment Before and after treatment Lactobacillus Staphylococcus Enterobacter Yeast 7.16±1.124.54±1.153.40±0.544.23±1.25 5.12±1.445.37±0.705.15±1.025.82±0.76 7.99±0.713.63±0.553.27±0.663.84±0.60 <0.001<0.001<0.001<0.001 <0.001<0.001>0.5>0.2 <0.001<0.001<0.001<0.05

5. The therapeutic effect of using this preparation on non-specific vaginitis. The results in Table 5 show that after inoculation of this preparation, the patient's symptoms are improved

Table 4 Therapeutic effect of using this preparation on non-specific vaginitis case number Before treatment After treatment secretion pH Lactobacillus secretion pH Lactobacillus 12345678910 2++2+++2++++ 5.05.55.04.56.05.05.55.55.05.0 +--+---++- +--+-++++- 3.03.54.03.53.03.03.53.54.04.0 3+3+3+3+4+4+4+2+3+3+

6. See Table 6, Table 7, Table 8, and Table 9 for changes in vaginal indicators before and after treatment of bacterial vaginosis using this preparation. The results showed that the pH value of the vagina before and after the medication was significantly lower than that before the medication, and there was a significant difference. Vaginal trichomoniasis, discharge, leucorrhea, and pruritus were significantly different before and after treatment, and were significantly better after treatment than before treatment.

Table 6 Comparison of changes in vaginal pH project Before medication after medication t P Vaginal pH 5.14±＞0.050.2630 cases (＞4.5) 4.35±0.3626(＜4.5) 9.66 <0.05*

*P value has significant difference

Table 7 Comparison of changes in Trichomonas vaginalis grading Before medication after medication u P -++++++Total 0217230 2910030 7.30 ＜0.01***

***P value has extremely significant difference

Table 8 Comparison of changes in vaginal itching grading Before medication after medication u P -+++Total 025430 1911030 6.08 ＜0.01***

***P value has extremely significant difference

Table 9 Comparison of changes in vaginal discharge and leucorrhea grading Before medication after medication u P -+++Total 0171330 1416030 6.38 ＜0.01***

***P value has extremely significant difference

Example 1 5g of Lactobacillus delbrueckii, 10g of Lactobacillus acidophilus, 700g of lactose, 100g of stachyose, fully mixed, containing 5-1.5 billion/g of viable probiotic bacteria, subpackaged into capsules, and packaged. External vaginal use, once a day, one capsule each time. The preparation process of Lactobacillus delbrueckii and Lactobacillus acidophilus bacteria powder is as follows:

(1) Preparation of Lactobacillus delbrueckii bacteria powder: Insert the seed liquid of Lactobacillus delbrueckii strain into sterilized 2% glucose by weight, 1.5% peptone, 1% yeast extract, 0.5 % sodium acetate, 0.5% ammonium citrate medium, after anaerobic fermentation at 36°C-39°C, the bacteria were centrifuged, and the bacteria accounting for 40% of the weight of the protection solution were added to the sodium glutamate containing 4% by weight The Lactobacillus delbrueckii powder can be obtained after mixing with 5% high-fat milk powder freeze-drying protection solution and freeze-drying.

(2) Preparation of Lactobacillus acidophilus bacterium powder: Insert the seed solution of Lactobacillus acidophilus strain by weight 1% inoculum into medium containing 2% glucose by weight, 1.5% peptone, 1% yeast extract and pass through 32°C- After anaerobic fermentation at 36°C, the cells were separated by centrifugation, and the cells accounting for 45% of the weight of the protection solution were added to the freeze-dried protection solution of 150mM trehalose, acid neutralizing substances, and antioxidant substances, mixed, and left for 2 hours. After freeze-drying, the Lactobacillus acidophilus bacteria powder can be obtained.

Example 2 10 g of Lactobacillus delbrueckii, 15 g of Lactobacillus acidophilus, 500 g of lactose, and 300 g of trehalose, mixed thoroughly, containing 5-1.5 billion viable probiotic bacteria/g, were divided into capsules and packaged. Take orally, 3 times a day, 2-4 capsules each time. The preparation process of Lactobacillus delbrueckii and Lactobacillus acidophilus bacteria powder is as follows:

(1) Preparation of Lactobacillus delbrueckii bacteria powder: Insert the seed liquid of Lactobacillus delbrueckii strain into sterilized 3% glucose by weight, 0.5% peptone, 0.8% yeast extract, 0.35 % sodium acetate, 0.35% ammonium citrate medium, after anaerobic fermentation at 36°C-39°C, the cells were centrifuged to separate the cells, and the cells accounting for 32% of the weight of the protection solution were added to the sodium glutamate containing 3% by weight The Lactobacillus delbrueckii powder can be obtained after mixing with 10% high-fat milk powder freeze-drying protection solution and freeze-drying.

(2) Preparation of Lactobacillus acidophilus bacterium powder: Insert the seed liquid of Lactobacillus acidophilus strain by weight 3% into the culture medium containing 3% glucose by weight, 0.5% peptone and 0.5% yeast extract, through 32°C- After anaerobic fermentation at 36°C, the cells were separated by centrifugation, and the cells accounting for 38% of the weight of the protection solution were added to the freeze-dried protection solution of 200mM trehalose, acid neutralizing substances and antioxidant substances, mixed, and left for 3 hours , and the Lactobacillus acidophilus powder can be obtained after freeze-drying.

Example 3 15g of Lactobacillus delbrueckii, 5g of Lactobacillus acidophilus, 600g of lactose, 200g of trehalose, add 500g of effervescent powder, fully mix and contain 5-1.5 billion viable probiotic bacteria/g, compress into tablets, pack . External vaginal use, once a day, one tablet each time. The preparation process of Lactobacillus delbrueckii and Lactobacillus acidophilus bacteria powder is as follows:

(1) Preparation of Lactobacillus delbrueckii bacteria powder: Insert the seed liquid of Lactobacillus delbrueckii strain into sterilized 1% glucose by weight, 0.8% peptone, 0.5% yeast extract, 0.2 % sodium acetate, 0.2% ammonium citrate medium, after anaerobic fermentation at 36°C-39°C, centrifuge to separate the bacteria, add 25% of the weight of the protection solution to the bacteria containing 5% of the weight of sodium glutamate The Lactobacillus delbrueckii bacteria powder can be obtained after mixing with 15% high-fat milk powder freeze-drying protection solution and freeze-drying.

(2) Preparation of Lactobacillus acidophilus bacterium powder: Insert the seed solution of Lactobacillus acidophilus strain by weight 5% inoculum into medium containing 1% glucose by weight, 0.8% peptone, 0.8% yeast extract through 32°C- After anaerobic fermentation at 36°C, the cells were separated by centrifugation, and the cells accounting for 30% of the weight of the protection solution were added to the freeze-dried protection solution of 100mM trehalose, acid neutralizing substances, and antioxidant substances, mixed, and left for 4 hours. After freeze-drying, the Lactobacillus acidophilus bacteria powder can be obtained.

Claims (2)

1, a kind of membrane-strengthening agent preparation that is used to adjust flora prevention or treatment bacterial vaginosis is characterized in that: Deshi Lactobacillus 5-15 part is by weight ratio arranged, and bacillus acidophilus 5-15 part, lactose 500-700 part, stachyose 100-300 part,

2, a kind of membrane-strengthening agent preparation process thereof that is used to adjust flora prevention or treatment bacterial vaginosis, it is characterized in that: get Deshi Lactobacillus 5-15 part by weight ratio, bacillus acidophilus 5-15 part, lactose 500-700 part, stachyose 100-300 part, contain number of live bacteria of probiotics 5-15 hundred million/g after the mixing, divide encapsulated or compressed tablets, preparation technology is as follows for Deshi Lactobacillus, bacillus acidophilus's mycopowder:

(1) preparation of Deshi Lactobacillus mycopowder: containing in weight 1%-3% glucose, 0.5-1.5% peptone, 0.5-1% yeast extract, 0.2-0.5% sodium acetate, the 0.2-0.5% ammonium citrate culture medium after the access of 1-5% inoculum concentration has been sterilized by weight with Deshi Lactobacillus bacterial strain seed liquor; behind 36 ℃ of-39 ℃ of anaerobic fermentations; centrifugalize goes out thalline, the thalline that accounts for protection liquid weight 25-40% is added in the sodium glutamate that contains weight 3-5% and the 5-15% greasiness milk powder frozen-dried protective liquid can obtain the Deshi Lactobacillus mycopowder after the mixed freezing drying.

(2) preparation of bacillus acidophilus's mycopowder: with the strain of lactobacillus acidophilus seed liquor by weight the 1-5% inoculum concentration insert and to contain in weight 1%-3% glucose, 0.5-1.5% peptone, the 0.5-1% yeast extract culture medium behind 32 ℃ of-36 ℃ of anaerobic fermentations; centrifugalize goes out thalline; add the thalline that accounts for protection liquid weight 30-45% in 100-200mM trehalose and the acid and in the frozen-dried protective liquid of material and antioxidant etc.; mix; placed 2-4 hour, and can obtain bacillus acidophilus's mycopowder after the lyophilization.