CN106692204B - Probiotics composition for relieving or improving pelvic inflammation and preparation method thereof - Google Patents
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Abstract
The invention discloses a probiotic composition for relieving or improving pelvic inflammation, which consists of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum; in the probiotic composition, the total amount of probiotics is 8.0 × 109‑4.0×1010cfu/g. The probiotic composition is also prepared into a probiotic traditional Chinese medicine preparation by a synergistic formula of the probiotic composition and a medicine-food homologous traditional Chinese medicine. Furthermore, the invention prepares a product for relieving or improving pelvic inflammation by compounding the probiotic composition, the medicinal and edible traditional Chinese medicine extract, the prebiotics and the like. The probiotic composition can enhance the immunity of organisms, promote the development of physiological normal probiotic groups in the genital tract, indirectly control and eliminate pathogenic bacteria through biological antagonism so as to regulate the micro-ecological balance in the genital tract, thereby realizing the relief or improvement of pelvic inflammation.
Description
Technical Field
The invention relates to the field of biomedicine, in particular to a probiotic composition for relieving or improving pelvic inflammation and a preparation method thereof.
Background
Pelvic Inflammatory Disease (PID) is a group of diseases caused by inflammation of the female upper genital tract, including endometritis, salpingitis, salpingo-ovarian abscess and pelvic peritonitis. Inflammation may be localized to one site or may involve several sites simultaneously, with salpingitis and salpingo-oophoritis being the most common.
Pelvic inflammation is a common disease which endangers the health of women, and endogenous opportunistic infection caused by the ascending of inherent bacteria in the vagina due to the microecological disorder (dysiosis) of the vagina is a main reason of the pelvic inflammation. The main resident bacteria in the reproductive tract include lactobacillus, staphylococcus epidermidis, colibacillus, corynebacterium, streptococcus faecalis, mycoplasma, candida albicans, enterococcus digesta, bacteroides and the like. The lactobacillus is the most important resident bacterium with the largest quantity in the normal flora of the vagina, accounts for more than 95 percent and is the normal dominant bacterium. Not only are probiotics closely related to gastrointestinal tract health, but certain probiotic strains are also beneficial to female genitourinary system health, and can prevent and treat female genitourinary system infection such as bacterial vaginitis, pelvic inflammatory disease and other related complications. The liquid in the healthy pelvic cavity is little, the liquid in the pelvic cavity is obviously increased when the pelvic cavity is in inflammatory lesion, most of the pelvic inflammatory diseases are caused by bacteria in the vagina ascending along a mucous membrane to infect pelvic organs, a large amount of anaerobic bacteria exist in the normal vagina, and once the environment is changed or the conditions are favorable, such as local blood supply insufficiency; presence of damaged or necrotic tissue; presence of foreign bodies such as intrauterine devices; exogenous microbes grow and reproduce to cause tissue damage, bacteria in the vagina can flow upwards to the uterine cavity, the oviduct and other parts along the mucous membrane, and good anaerobic conditions are provided for the growth and the reproduction of anaerobic bacteria to cause inflammation, and the intensity of inflammatory reaction can be reflected by the expression level of some cell factors.
The cytokine is a polypeptide protein which is synthesized and secreted by immune active cells and the like and mediates inflammation and immune response, is produced by lymphocytes, monocytes, macrophages, fibroblasts and the like, plays an important role in host tissue damage resisting process, relates to a plurality of physiological and pathological complex processes in organisms, can activate lymphocytes, regulate the growth and differentiation of lymphocytes and other cells, enhance immune response and metabolism and functions of inflammatory cells, and is closely related to the occurrence and outcome of a plurality of clinically important diseases.IL-2 is a cytokine with various biological activities, is mainly secreted by helper T cells and macrophages, has the functions of regulating immunity and increasing body anti-infective ability.As an important cytokine, IL-2 is combined with IL-2 receptors on the surfaces of T cells, B cells and monocytes, can cause T cell proliferation, can promote cytotoxic T cell killing, enhance natural killer cell activity, can promote immunoglobulin secretion of B cells such as IgG and the like, thus has important significance in immune regulation, IL-2 level, reflects that the T cell activation from a side surface and the IL-2 receptor activity of lymphocytes can strongly stimulate inflammatory cell proliferation, promote inflammatory cell proliferation of endothelial cells, promote inflammatory cell proliferation and promote inflammatory cell proliferation of endothelial cells to generate excessive inflammatory cell proliferation, so that TNF-6 and TNF-2 can cause excessive inflammatory cell infiltration, and promote inflammatory cell proliferation of immune cell proliferation, and inflammatory cell proliferation, so that the inflammatory cell proliferation of endothelial cell proliferation and inflammatory cell proliferation of endothelial cell receptor can cause the inflammatory cell proliferation, and inflammatory cell proliferation.
The current treatment methods for pelvic inflammatory diseases mainly comprise: antibiotic therapy, traditional Chinese medicine fumigation therapy, microecology therapy and the like, wherein the antibiotic is the most widely applied medicine in clinical treatment of infectious diseases at present. However, the antibiotic destroys the normal flora of the vagina while inhibiting pathogenic bacteria, changes the advantages of microorganisms, generates a large amount of pathogenic microorganisms with drug resistance, and seriously influences the curative effect of the antibiotic once infected again.
The fumigation and washing therapy of traditional Chinese medicine directly acts on the affected part by local stimulation, eliminates symptoms and solves focus. The medicine can be wholly regulated through the absorption of the skin, orifices and acupoints on the smell of the medicine, the regulation of the channels and collaterals, the whole pharmacological effect caused by the internal organ distribution and the stimulation of the medicine, so that the medicine can play a role and promote the relative balance of yin and yang. Modern physicians mostly adopt the method of clearing heat, removing dampness and removing blood stasis, but some patients have deficiency-cold, and use bitter-cold products or abuse antibiotics for a long time to damage yang qi, resulting in internal excess of yin-cold, congealing cold and blood stasis.
Micro-ecological therapy: the essence of the method is a prevention and treatment method aiming at supporting normal flora, antagonizing pathogenic bacteria or conditioned pathogenic bacteria and adjusting and restoring microecological balance. The microecological regulator is a product of probiotics (microorganisms) and metabolites thereof and growth promoting substances which are guided by the microecological theory, correct the unbalance of the microecological, keep the balance of the microecological, and improve the health level of organisms or the health state.
With the intensive research on reproductive tract infection RTI (regenerative track infection), people have gradually realized that the ecological balance of normal vaginal flora is maintained, and the dominant position of the beneficial vaginal bacterium, namely lactobacillus, is an effective method for preventing and treating reproductive tract infectious diseases. Currently, micro-ecological therapy is gaining attention from medical workers as the toxic side effects of antibiotics and abuse of antibiotics lead to the emergence of a large number of drug-resistant flora and superinfection, making the dominance of antibiotics in the treatment of infectious diseases more and more challenging. Therefore, if the microecological regulator can be widely applied to clinic, a new way for preventing and treating the genital tract infection of women is necessarily developed, and safe and effective good health care medicines are also provided for the vast women. However, no report about the effect of the probiotics on relieving or improving pelvic inflammation exists at present.
Disclosure of Invention
In view of the above-mentioned deficiencies of the prior art, the present invention aims to provide a probiotic composition for relieving or improving pelvic inflammation. The probiotic composition can enhance the immunity of organisms, promote the development of physiological normal probiotic groups in the genital tract, indirectly control and eliminate pathogenic bacteria through biological antagonism so as to regulate the micro-ecological balance in the genital tract, thereby realizing the relief or improvement of pelvic inflammation.
The other purpose of the invention is to provide a probiotic traditional Chinese medicine preparation for relieving or improving pelvic inflammation, which achieves the purpose of treating while regulating by combining and synergizing the probiotics and the traditional Chinese medicine, and obviously enhances the treatment effect on the pelvic inflammation.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect of the invention, a probiotic composition for relieving or improving pelvic inflammation is provided, which consists of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum;
in the probiotic composition, the total amount of probiotics is 8.0 × 109-4.0×1010cfu/g。
Preferably, in the probiotic composition, the mass ratio of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum is 2:1: 1.
More preferably, the Lactobacillus acidophilus is Lactobacillus acidophilus (Lactobacillus acidophilus) BLCC2-0024 with the preservation number of CCTCC NO: M2015129; the lactobacillus plantarum is lactobacillus plantarum LP with a preservation number of CCTCC M2010150.
The invention also provides a preparation method of the probiotic composition, which comprises the following steps:
(1) respectively activating Lactobacillus rhamnosus, Lactobacillus acidophilus and Lactobacillus plantarum with culture medium, inoculating into fermentation culture solution, and standing at 37 deg.C to make the thallus concentration reach 4.0 × 10 in logarithmic phase9-8.0×109cfu/ml；
(2) Respectively centrifuging the fermentation liquor, and removing the supernatant to obtain bacterial sludge of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum;
(3) and (3) uniformly mixing the bacterial sludge and the freeze-drying protective agent, and freeze-drying to obtain the microbial inoculum.
The second aspect of the invention provides a probiotic traditional Chinese medicine preparation for relieving or improving pelvic inflammation, which is prepared by synergistically combining the probiotic composition and a medicinal and edible traditional Chinese medicine extract with the effect of treating pelvic inflammation.
The medicinal and edible traditional Chinese medicine extract is prepared from the following raw material medicines in parts by weight:
2-3 parts of angelica, 2-5 parts of peach kernel, 6-12 parts of raw hawthorn, 10-15 parts of poria cocos, 2-3 parts of cinnamon, 3-9 parts of liquorice, 3-10 parts of rose, 6-10 parts of dandelion and 15-30 parts of houttuynia cordata.
Preferably, the medicinal and edible traditional Chinese medicine extract is prepared from the following raw material medicines in parts by weight:
2 parts of angelica, 3 parts of peach kernel, 8 parts of raw hawthorn, 10 parts of tuckahoe, 2 parts of cinnamon, 6 parts of liquorice, 6 parts of rose, 6 parts of dandelion and 15 parts of houttuynia cordata.
The invention also provides a preparation method of the medicinal and edible traditional Chinese medicine extract, which comprises the following steps:
soaking radix Angelicae sinensis, semen Persicae, fructus crataegi, Poria, cortex Cinnamomi, Glycyrrhrizae radix, flos Rosae Rugosae, herba Taraxaci, and herba Houttuyniae in 8-10 times of water for 30-60min, decocting for 20-40min after water is boiled, and filtering to obtain first filtrate; decocting the residue with 6-8 times of water for two times, decocting for 20-40min after the water is boiled, filtering to obtain second filtrate, mixing with the first filtrate, concentrating until the solid content is above 30%, spray drying, pulverizing, and sieving to obtain extract of Chinese medicinal materials with homology of medicine and food;
preferably, in the probiotic traditional Chinese medicine preparation, the weight ratio of the probiotic composition to the medicinal and edible traditional Chinese medicine extract is (2-10): (20-30).
The application of the probiotic composition and/or the probiotic traditional Chinese medicine preparation in preparing a product for relieving or improving pelvic inflammation is also within the protection scope of the invention.
In the application, the product is a medicament, food or health-care product.
The third aspect of the invention provides a product for relieving or improving pelvic inflammation, which comprises the following components in percentage by mass:
2-10% of probiotic composition, 20-30% of medicinal and edible traditional Chinese medicine extract, 50-60% of prebiotics and 5-10% of auxiliary material filler.
The prebiotics are selected from one or more of isomaltooligosaccharide, fructo-oligosaccharide and xylo-oligosaccharide; preferably, the prebiotics consist of isomaltooligosaccharide, fructooligosaccharide and xylooligosaccharide in a mass ratio of 5:3: 1.
The auxiliary material filling is selected from one or more of maltodextrin, citric acid, fruit powder and sugar alcohol.
The invention also provides a preparation method of the product for relieving or improving pelvic inflammation, which comprises the following steps:
(1) preparing the probiotic composition and the medicinal and edible traditional Chinese medicine extract according to the method;
(2) mixing probiotic composition, medicinal and edible Chinese medicinal extract, prebiotics and adjuvant filler uniformly to obtain the final product.
The invention has the beneficial effects that:
(1) the probiotic composition with the function of relieving or improving pelvic inflammation can enhance the immunity of organisms, promote the development of physiological normal probiotic groups in the genital tract, indirectly control and eliminate pathogenic bacteria through biological antagonism so as to regulate the micro-ecological balance in the genital tract, thereby relieving or improving the pelvic inflammation.
(2) The probiotic traditional Chinese medicine preparation provided by the invention is prepared by the synergistic combination of the probiotic composition and the medicinal and edible traditional Chinese medicine with the effect of treating pelvic inflammation, wherein an extract of the medicinal and edible traditional Chinese medicine plays a role in strengthening body resistance and consolidating the constitution and eliminating pathogenic factors as a micro-ecological regulator, thereby promoting the recovery of lactobacillus in the genital tract, improving the immunity of the organism, and killing and inhibiting pathogenic microorganisms to regulate dysbacteriosis in the genital tract; the probiotics can better decompose the traditional Chinese medicine extract into small molecular substances, so that the intestinal absorption is facilitated, the traditional Chinese medicine and the probiotics are reasonably compatible, and experiments prove that the aim of treating while regulating can be fulfilled, the effect of enhancing is achieved, and the perfect combination of the probiotics and the traditional Chinese medicine is realized.
(3) The product for relieving or improving pelvic inflammation is prepared from the probiotic composition, a medicine-food homologous traditional Chinese medicine, prebiotics and an auxiliary material filler, wherein the prebiotic component is added on the basis of the synergistic effect of the probiotic and the traditional Chinese medicine, and the prebiotics are used as a dietary supplement and can selectively stimulate the growth and activity of bacteria in one or a small number of colonies to have beneficial influence on a host. The added prebiotics component can generate further synergistic effect with the probiotic composition, thereby being more beneficial to the improvement and the relief of pelvic inflammation symptoms.
(4) The product for relieving or improving pelvic inflammation has the advantages of simple preparation process, less loss of each effective component in the traditional Chinese medicine, high safety and guarantee of stability among product batches.
Detailed Description
It should be noted that the following detailed description is exemplary and is intended to provide further explanation of the disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs.
It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of example embodiments according to the present application. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of stated features, steps, operations, devices, components, and/or combinations thereof, unless the context clearly indicates otherwise.
As described in the background art, the prior art lacks a micro-ecological regulator capable of relieving or improving pelvic inflammation, and in order to solve the technical problems, the application provides a probiotic composition for relieving or improving pelvic inflammation, which can enhance the immunity of the organism, promote the development of physiologically normal probiotic flora in the genital tract, indirectly control and eliminate pathogenic bacteria through biological antagonism so as to regulate the micro-ecological balance in the genital tract, thereby relieving or improving pelvic inflammation.
In one embodiment of the present application, there is provided a probiotic composition for relieving or improving pelvic inflammation, consisting of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum; in the probiotic composition, the total amount of probiotics is 8.0 × 109-4.0×1010cfu/g。
As a preferred embodiment, the probiotic composition has a mass ratio of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum of 2:1: 1.
As the performances of strains of the same species from different sources are greatly different, the application also adjusts the types of probiotics in the probiotic composition, the probiotics of the same species but different separation sources and the combination thereof in the test process, and as a result, the effect of compounding Lactobacillus acidophilus (Lactobacillus acidophilus) BLCC2-0024 with the preservation number of M2015129 and Lactobacillus plantarum LP and Lactobacillus rhamnosus with the preservation number of CCTCC M2010150 is the best compared with the other types or sources of probiotic combinations.
The Lactobacillus acidophilus (Lactobacillus acidophilus) BLCC2-0024 with the preservation number of M2015129 has been preserved in the China center for type culture collection at 3.18.2015, and the patent of 'a blood sugar reducing probiotic preparation and a preparation method thereof' (CN105039217A) is recorded to constitute the prior art, so that the preservation evidence is not provided any more. The functions of the strain at present mainly comprise reducing blood sugar and relieving yin deficiency type constipation of the old, and reports on the strain for treating pelvic inflammation are not available.
The lactobacillus plantarum LP with the preservation number of CCTCC M2010150 is already preserved in the China Center for Type Culture Collection (CCTCC) in 2010, 6 and 21, and the patent is recorded in 'a lactobacillus capable of being used for biological preservation and freshness preservation and application thereof' (CN101914475A), which constitutes the prior art, so that no preservation evidence is provided. The functions related to the strain at present are mainly used for the preservation and freshness of fruits, and reports on the use of the strain in treating pelvic inflammation are not found.
The lactobacillus rhamnosus is a conventional commercial product.
The three selected strains can promote the development of physiological normal probiotic groups in the genital tract, and indirectly control and eliminate pathogenic bacteria such as escherichia coli, staphylococcus aureus and the like through biological antagonism so as to adjust the micro-ecological balance in the genital tract; meanwhile, the health-care product can enhance the immunity of the organism, regulate the intestinal system and play the health-care and body-building effects, thereby realizing the relief or improvement of pelvic inflammation.
In another embodiment of the application, a probiotic traditional Chinese medicine preparation for relieving or improving pelvic inflammation is provided, and the probiotic traditional Chinese medicine preparation is prepared by synergistically combining the probiotic composition and a medicinal and edible traditional Chinese medicine extract with the effect of treating pelvic inflammation.
The medicinal and edible traditional Chinese medicine extract is prepared from the following raw material medicines in parts by weight:
2-3 parts of angelica, 2-5 parts of peach kernel, 6-12 parts of raw hawthorn, 10-15 parts of poria cocos, 2-3 parts of cinnamon, 3-9 parts of liquorice, 3-10 parts of rose, 6-10 parts of dandelion and 15-30 parts of houttuynia cordata.
The preparation method of the medicinal and edible traditional Chinese medicine extract comprises the following steps:
soaking radix Angelicae sinensis, semen Persicae, fructus crataegi, Poria, cortex Cinnamomi, Glycyrrhrizae radix, flos Rosae Rugosae, herba Taraxaci, and herba Houttuyniae in 8-10 times of water for 30-60min, decocting for 20-40min after water is boiled, and filtering to obtain first filtrate; decocting the residue with 6-8 times of water for two times, decocting for 20-40min after the water is boiled, filtering to obtain second filtrate, mixing with the first filtrate, concentrating until the solid content is above 30%, spray drying, pulverizing, and sieving to obtain extract of Chinese medicinal materials with homology of medicine and food.
As a preferable embodiment, in the probiotic traditional Chinese medicine preparation, the weight ratio of the probiotic composition to the medicinal and edible traditional Chinese medicine extract is (2-10): (20-30).
The traditional Chinese medicine contains sugar, fat, amino acid and trace elements, and can provide necessary nutrient substances for organisms. Animal experiments and clinical observation prove that part of the Chinese herbal medicines have the functions of supporting the growth of normal flora, adjusting the imbalance of the flora and improving the planting resistance, and are ideal micro-ecological regulators. The traditional Chinese medicine can play roles in strengthening body resistance and consolidating constitution and eliminating pathogenic factors, not only promotes the recovery of lactobacillus in vagina, improves the immunity of organisms, but also kills and inhibits pathogenic microorganisms to regulate vaginal dysbacteriosis flora, which is probably one of the reasons that the curative effect is more stable and the recurrence is not easy. The traditional Chinese medicine preparation is used as a micro-ecological regulator and is combined with a live bacteria preparation, so that the purpose of treating and regulating at the same time is achieved, and the effect is enhanced.
The formula of the medicinal and edible traditional Chinese medicine extract comprises the following components:
the angelica has the effects of tonifying blood and blood, regulating menstruation, relieving pain, moistening dryness, lubricating intestines, resisting cancer, resisting aging, resisting immunity and relieving the abdominal pain caused by pelvic inflammation; the peach kernel has the effects of activating blood circulation to dissipate blood stasis, relaxing bowel, relieving cough and asthma; the hawthorn has good medicines for promoting digestion, removing stagnation, promoting blood circulation and reducing phlegm; the tuckahoe has the effects of promoting diuresis and eliminating dampness, strengthening spleen and calming heart; cortex Cinnamomi has effects of invigorating primordial yang, warming spleen and stomach, removing cold accumulation, and promoting blood circulation; the licorice root has the functions of mainly clearing away heat and toxic material, treating abdominal diseases and other symptoms; rose is sweet in nature, slightly bitter in taste and warm in nature. It enters liver and spleen meridians. Has effects in promoting qi circulation, relieving depression, regulating blood circulation, and relieving pain; the dandelion has the efficacy of clearing away heat and toxic materials, and is a good medicine for gynecological diseases; yu xing Cao is pungent in flavor and cold in nature and enters lung meridian. Has effects in clearing away heat and toxic materials, relieving swelling, treating sore, promoting urination, removing dampness, clearing away heat, relieving dysentery, invigorating stomach, and promoting digestion; the whole formula has the effects of nourishing blood, removing blood stasis, promoting blood circulation, removing toxin, dredging channels and collaterals, regulating qi, tonifying spleen and strengthening physique, and fundamentally relieves the pelvic inflammation symptoms.
The probiotic composition can better decompose the traditional Chinese medicine extract into small molecular substances, and is beneficial to the absorption of intestinal tracts, so that the perfect combination of the probiotics and the traditional Chinese medicine is realized. In the probiotic traditional Chinese medicine preparation, the traditional Chinese medicine components with homology of medicine and food and the probiotic components are reasonably compatible, and experiments prove that the purpose of treating while regulating can be achieved, and the effect is enhanced.
Through analysis, the probiotic traditional Chinese medicine preparation disclosed by the invention has the effect of relieving or improving pelvic inflammation, and can inhibit TNF- α and IL-6 mediated chemotactic reaction by improving the immunity of an organism, regulating the expression of serum cytokines of an inflammatory organism and regulating the expression of local inflammatory factors of pelvic tissues, and relieve the inflammatory injury of the organism tissues by increasing the expression of IL-2 to relieve the inflammatory reaction, so that the effect of relieving or improving the pelvic inflammation is achieved.
In another embodiment of the application, a product for relieving or improving pelvic inflammatory disease is provided, which comprises the following components in percentage by mass:
2-10% of probiotic composition, 20-30% of medicinal and edible traditional Chinese medicine extract, 50-60% of prebiotics and 5-10% of auxiliary material filler.
The prebiotics are selected from one or more of isomaltooligosaccharide, fructo-oligosaccharide and xylo-oligosaccharide; preferably, the prebiotics consist of isomaltooligosaccharide, fructooligosaccharide and xylooligosaccharide in a mass ratio of 5:3: 1.
The auxiliary material filling is selected from one or more of maltodextrin, citric acid, fruit powder and sugar alcohol.
The invention also provides a preparation method of the product for relieving or improving pelvic inflammation, which comprises the following steps:
(1) preparing the probiotic composition and the medicinal and edible traditional Chinese medicine extract according to the method;
(2) mixing probiotic composition, medicinal and edible Chinese medicinal extract, prebiotics and adjuvant filler uniformly to obtain the final product.
In order to make the technical solutions of the present application more clearly understood by those skilled in the art, the technical solutions of the present application will be described in detail below with reference to specific embodiments.
The test materials used in the examples of the present invention are all conventional in the art and commercially available.
Example 1: preparation of freeze-dried powder of probiotics
1) Activating three strains of Lactobacillus acidophilus (Lactobacillus acidophilus) BLCC2-0024 with preservation number of CCTCC NO: M2015129 and Lactobacillus plantarum LP and Lactobacillus rhamnosus with preservation number of CCTCC M2010150 with culture medium, inoculating into fermentation culture solution, and standing at constant temperature of 37 deg.C for culture to make the thallus concentration reach 4.0 × 10 in logarithmic growth phase9-8.0×109cfu/ml；
Preparation of lactobacillus seed liquid: taking out the strain storage tube, streaking with lactobacillus slant solid culture medium respectively for resuscitation, and culturing at 37 deg.C for 24-36 hr. And selecting colony crystal violet staining microscopic on the slant culture medium, wherein the colony is Brevibacterium or Long bacillus. Colonies were picked and inoculated into 100mL of lactic acid bacteria medium and cultured in a 37 ℃ incubator for 24 h. Inoculating the seeds with an inoculum size of 2-5% into a 3L large triangular flask, culturing at 37 ℃ for 24-36h, detecting the concentration of a bacterial liquid, and inoculating the liquid bacterial strain with the bacterial content of more than 20 hundred million/ml, thereby being capable of being used as a lactobacillus liquid strain for inoculation. Wherein the MRS culture medium: 2.0% of glucose, 0.2% of ammonium citrate, 0.5% of sodium acetate, 0.5% of dipotassium phosphate, 0.02% of manganese sulfate, 0.05% of magnesium sulfate, 1.0% of peptone, 1.0% of beef extract, 0.5% of yeast extract and 800.1% of tween, and adjusting the pH value to 6.0; 1.5% agar powder was added to the solid medium. The culture medium sterilization conditions were: sterilizing at 121 deg.C under 0.10-0.15MPa for 20 min.
2) Centrifuging various fermentation liquors obtained in the step 1) respectively, and discarding supernatant to obtain bacterial sludge;
3) preparing a freeze-drying protective agent; the formula of the freeze-drying protective agent comprises the following components: 25g of skimmed milk powder, 8g of cane sugar, 1g of edible glycerin and 100mL of distilled water.
4) Mixing the bacterial sludge and the freeze-drying protective agent uniformly, pre-freezing the mixture, and then carrying out vacuum freeze-drying to prepare the freeze-dried powder of the active probiotics with the coating layer.
Example 2: preparation of medicinal and edible traditional Chinese medicine extract
1) According to the preferable weight portions: 2g of angelica, 3g of peach kernel, 8g of raw hawthorn, 10g of tuckahoe, 2g of cinnamon, 6g of liquorice, 6g of rose, 6g of dandelion and 15g of houttuynia cordata;
2) soaking the mixture in 10 times of water for 40min, decocting for 30min after the water is boiled, and filtering to obtain a first filtrate; decocting the residue with 8 times of water for two times, decocting for 30min after the water is boiled, filtering to obtain a second filtrate, mixing with the first filtrate uniformly, concentrating the Chinese medicinal liquid by a vacuum reduced pressure rotary evaporator until the solid content is above 30%, feeding into a spray dryer, controlling the air inlet temperature at 140 deg.C, the air outlet temperature at 90 deg.C, the rotation speed of an atomizer at 45 r/min and the feeding speed at 5 ml/min to obtain spray dried powder, pulverizing, and sieving with a sieve of above 100 meshes to obtain the extract of the medicine and food homologous Chinese medicinal materials.
Example 3: preparation of probiotic composition for relieving or improving pelvic inflammation
The probiotic freeze-dried powder prepared in the embodiment 1, namely 3 kinds of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum are scientifically compounded according to the mass ratio of 2:1:1 and uniformly mixed to obtain the probiotic composition capable of relieving or improving pelvic inflammation.
Example 4: preparation of probiotic traditional Chinese medicine preparation for relieving or improving pelvic inflammation
Respectively sieving the probiotic composition prepared in example 3 and the medicinal and edible traditional Chinese medicine extract prepared in example 2 by a sieve of more than 100 meshes, and weighing for later use, wherein the mass ratio of the probiotic composition to the medicinal and edible traditional Chinese medicine extract is 8: and 30, batching. Mixing the above materials in a mixer to obtain probiotic Chinese medicinal preparation with effect in relieving or improving pelvic inflammatory disease.
Example 5: preparation of product for relieving or improving pelvic inflammation
Respectively sieving the freeze-dried powder of the probiotics prepared in the embodiment 1 and the extract of the medicine-food homologous traditional Chinese medicine prepared in the embodiment 2 by a sieve of more than 100 meshes, and weighing for later use; the probiotics powder is 8 percent, the traditional Chinese medicine extract is 30 percent, the prebiotics is 55 percent and the auxiliary material filler is 7 percent. The above materials are fully and uniformly mixed in a mixer until the color is uniform and no color difference exists, and the product with the function of relieving or improving pelvic inflammation is obtained.
The prebiotics are composed of isomaltooligosaccharide, fructo-oligosaccharide and xylo-oligosaccharide in a mass ratio of 5:3: 1.
The auxiliary material filler is citric acid.
Example 6: improving effect on pelvic inflammation symptoms of pelvic inflammation model mice
1) Establishment of pelvic inflammatory disease mouse animal model
Animal preparation: a Kunming white mouse with an anogenital tract disease is selected to construct a mouse pelvic inflammation model.
Opening the abdomen of the animal: 4% chloral hydrate was intraperitoneally injected at 1% of the weight of the mouse, general anesthesia was performed, the mouse was placed on a fixed table several minutes later, the abdominal hair was subtracted, the skin was sterilized with 75% ethanol, the lower abdominal skin and muscle layer of the mouse were cut with a sterilized scissors forceps, and the abdominal cavity was exposed by cutting an incision of about 1 cm.
Uterus damage: exposing uterus, pricking the middle part of uterine horn at one side of uterus with a sterile needle head about 5-6 needles, closing abdomen layer by layer, and sterilizing the abdominal operation area.
2) Influence of probiotic composition with effect of relieving or improving pelvic inflammation on various visceral indexes of pelvic inflammation model mice
The mice selected from the above model were randomly divided into blank control group (without any treatment), sham operation group (with only open abdomen operation and without uterus treatment), model group, and probiotic group (prepared in example 3) (5.0 × 10)8cfu/d), probiotic traditional medicine (prepared in example 4) (0.025g/d), product group (prepared in example 5) (0.03g/d), commercial product group (0.037g/d), 10 per group; except for the blank control, the sham operation group and the model group, corresponding test samples were administered with an equal volume of pure water, and the gavage was continued for 21 d.
The next day after stopping the drug, mice were weighed, serum was taken and sacrificed, after dissection, liver, thymus and uterus were respectively weighed, and uterus was fixed with 4% neutral formaldehyde fixing solution, paraffin embedded and sectioned conventionally. HE staining, and observing the pathological morphological change of the uterus under a light microscope.
Table 1: effect on probiotic function of visceral index in mice (x + -s, n ═ 10)
Note: a, b, c indicate significant differences (0.01< P < 0.05).
As can be seen from Table 1, the uterus index shows that the model group has obvious swelling compared with the blank control group, the swelling degree is improved by 23.6 percent, which indicates that the molding is successful; compared with a model group, the uterus swelling degrees of a probiotic traditional Chinese medicine, a product group and a product group sold in the market in a tested sample are obviously improved, and the swelling degrees are respectively reduced by 12.1%, 18.0% and 17.5%, which shows that the application effect of the probiotic traditional Chinese medicine and the traditional Chinese medicine combination is obviously improved compared with the effect of the probiotic, and the product prepared in the embodiment 5 has a better effect, which shows that the product prepared by the invention has a certain improvement effect on the symptoms of pelvic inflammation; the thymus index shows that the model group has difference significance compared with the false operation group, but has no difference significance with other groups, which indicates that the inflammatory reaction of the model group influences the thymus index to a certain extent, so that the thymus index is increased compared with a blank control group; the liver index shows that the model group has difference and significance compared with the tested sample group, and the liver index of the tested group is reduced, which indicates that the probiotic composition, the probiotic traditional Chinese medicine preparation and the product have no damage or toxic effect on the liver and are safe.
3) Influence of probiotic composition with function of relieving or improving pelvic inflammation on uterine pathological changes of pelvic inflammation model mice
The uterus pathology observation indexes and integral standards are as follows: (1) occlusion or expansion of intrauterine adhesions: "-" no lesion score 0; lesions "+" < 1/3 scored 1; "+ +" 1/3-2/3 lesions scored 2; "+ + + +" 2/3 the lesions above were scored 3 points. (2) And (3) pathological changes of the cavity wall structure: "-" each layer structure is normally marked with 0 point; the + mucosal lamina glandular structure disappearance score 1; the boundary between the muscle layer and the mucosa layer is not marked for 2 minutes; the "+ + + +" hierarchy does not note 3 points. (3) Epithelial cell degeneration and necrosis: "-" monolayer columnar epithelium score 0; "+" epithelial cell flattening or shedding < 1/3 score 1; "+ +" epithelial flattening or shedding 1/3-2/3 lesions scored 2; "+ + + +" Whole layer epithelial cells degenerated bad death for 3 points. (4) Infiltration of chronic inflammatory cells: "-" no chronic inflammatory cell infiltration score 0; the "+" decimal fraction is scattered or focal and only marks 1 point in the inherent layer of the mucosa; "+ +" scattered or focal but deep into the muscle layer scored 2 points; the majority of +++, is dispersed or layered and covers 3 points of the whole layer. (5) Intimal congestion and edema: "-" No intimal congestion edema score 0; "+" mild hyperemia and edema of lamina propria score of 1; "+ +" clear hyperemia and edema score 2; the total hyperemia and edema score 3 points +++ "is given.
Table 2: influence on pathological change (scoring result) of mouse pelvic inflammation model uterus caused by mechanical injury
As can be seen from Table 2, the success rate of the model can reach more than 90%, and the degree of pathological changes of the uterus of the model group only given pure water is serious; the probiotic composition, the probiotic traditional Chinese medicine preparation, the product prepared in example 5 and the commercial product can obviously improve the pathological change degree of the uterus, and the product prepared in example 5 has the best effect and can be compared favorably with the commercial product, so that the pathological change degree is reduced by 65.5%.
4) The effect of probiotic compositions with the effect of relieving or improving pelvic inflammation on serum indicator cytokines of pelvic inflammation model mice table 3: influence on mouse pelvic inflammation model serum index cytokine caused by mechanical injury (x + -s, n is 10)
Note: a, b, c indicate significant differences (0.01< P < 0.05).
As can be seen from table 3, the content of the IL-2 model group is reduced by 6.20% compared with the blank control group, the content of the IL-6 and TNF- α model groups is respectively improved by 9.1% and 11.2% compared with the blank control group, and the content of TNF- α is significantly different between the two groups, which indicates that the modeling is successful, after the test sample is administered, the content of IL-2 is increased, the content of the probiotic group, the probiotic traditional Chinese medicine and the product prepared in example 5 are sequentially improved by 4.73%, 20.6% and 26.2% compared with the model group, although there is no significant difference compared with the commercial product group, the effect is better than that of the commercial product group, the content of the probiotic traditional Chinese medicine and the product prepared in example 6 are significantly different than that of the other groups, the content of the two groups is respectively reduced by 15.6% and 25.7%, and the content of the product group is significantly reduced by 21.8% compared with the commercial product group, which indicates that the product prepared in example 5 has a better effect than the commercial product, has a significant improvement effect on pelvic inflammation, and a synergistic effect on the pelvic inflammation caused by the reduction of the content of the probiotic product prepared in the probiotic composition and the probiotic composition, which has a synergistic effect of the beneficial bacteria, and a synergistic effect of the invention.
The above description is only a preferred embodiment of the present application and is not intended to limit the present application, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, improvement and the like made within the spirit and principle of the present application shall be included in the protection scope of the present application.
Claims (10)
1. A probiotic composition for relieving or improving pelvic inflammation, which is characterized by consisting of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum;
in the probiotic composition, the total amount of probiotics is 8.0 × 109-4 .0×1010cfu/g; the Lactobacillus acidophilus is Lactobacillus acidophilus (Lactobacillus acidophilus) BLCC2-0024 with the preservation number of CCTCC NO: M2015129; the lactobacillus plantarum is lactobacillus plantarum LP with a preservation number of CCTCC M2010150.
2. The probiotic composition according to claim 1, characterized in that the probiotic composition comprises lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum in a mass ratio of 2:1: 1.
3. A process for the preparation of the probiotic composition according to any of claims 1 to 2, characterized in that it comprises the following steps:
(1) respectively activating Lactobacillus rhamnosus, Lactobacillus acidophilus and Lactobacillus plantarum with culture medium, inoculating into fermentation culture solution, standing for culture to make thallus concentration reach 4.0 × 10 log phase9-8 .0×109cfu/ml；
(2) Respectively centrifuging the fermentation liquor, and removing the supernatant to obtain bacterial sludge of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum;
(3) and (3) uniformly mixing the bacterial sludge and the freeze-drying protective agent, and freeze-drying to obtain the microbial inoculum.
4. A probiotic traditional Chinese medicine preparation for relieving or improving pelvic inflammation, which is characterized by being prepared by synergistically combining the probiotic composition of claim 1 and a medicinal and edible traditional Chinese medicine extract with the effect of treating pelvic inflammation;
the medicinal and edible traditional Chinese medicine extract is prepared from the following raw material medicines in parts by weight:
2-3 parts of angelica, 2-5 parts of peach kernel, 6-12 parts of raw hawthorn, 10-15 parts of poria cocos, 2-3 parts of cinnamon, 3-9 parts of liquorice, 3-10 parts of rose, 6-10 parts of dandelion and 15-30 parts of houttuynia cordata.
5. The probiotic traditional Chinese medicine preparation according to claim 4, wherein the medicinal and edible traditional Chinese medicine extract is prepared from the following raw material medicines in parts by weight:
2 parts of angelica, 3 parts of peach kernel, 8 parts of raw hawthorn, 10 parts of tuckahoe, 2 parts of cinnamon, 6 parts of liquorice, 6 parts of rose, 6 parts of dandelion and 15 parts of houttuynia cordata.
6. The probiotic traditional Chinese medicine preparation for relieving or improving pelvic inflammation according to claim 4 or 5, which is characterized in that the extract of the medicine-food homologous traditional Chinese medicine is prepared by the following method:
soaking radix Angelicae sinensis, semen Persicae, fructus crataegi, Poria, cortex Cinnamomi, Glycyrrhrizae radix, flos Rosae Rugosae, herba Taraxaci, and herba Houttuyniae in 8-10 times of water for 30-60min, decocting for 20-40min after water is boiled, and filtering to obtain first filtrate; decocting the residue with 6-8 times of water for two times, decocting for 20-40min after the water is boiled, filtering to obtain second filtrate, mixing with the first filtrate, concentrating until the solid content is above 30%, spray drying, pulverizing, and sieving to obtain extract of Chinese medicinal materials with homology of medicine and food.
7. The probiotic traditional Chinese medicine preparation according to claim 4, wherein the weight ratio of the probiotic composition to the medicinal and edible traditional Chinese medicine extract is (2-10): (20-30).
8. Use of the probiotic composition of any one of claims 1-2 and/or the probiotic traditional Chinese medicine preparation of any one of claims 4-6 in the preparation of a product for relieving or improving pelvic inflammation.
9. A product for relieving or improving pelvic inflammation is characterized by comprising the following components in percentage by mass:
2-10% of probiotic composition, 20-30% of medicinal and edible traditional Chinese medicine extract, 50-60% of prebiotics and 5-10% of auxiliary material filler;
the prebiotics are selected from one or more of isomaltooligosaccharide, fructo-oligosaccharide and xylo-oligosaccharide;
the probiotic composition consists of lactobacillus rhamnosus, lactobacillus acidophilus and lactobacillus plantarum;
the lactobacillus acidophilus is lactobacillus acidophilus (Lactobacillus acidophilus) BLCC2-0024 with the preservation number of CCTCC NO: M2015129; the lactobacillus plantarum is lactobacillus plantarum LP with a preservation number of CCTCC M2010150;
the medicinal and edible traditional Chinese medicine extract is prepared from the following raw material medicines in parts by weight:
2-3 parts of angelica, 2-5 parts of peach kernel, 6-12 parts of raw hawthorn, 10-15 parts of poria cocos, 2-3 parts of cinnamon, 3-9 parts of liquorice, 3-10 parts of rose, 6-10 parts of dandelion and 15-30 parts of houttuynia cordata.
10. The product for relieving or improving pelvic inflammatory disease according to claim 9, wherein the prebiotic comprises isomalto-oligosaccharide, fructo-oligosaccharide and xylo-oligosaccharide in a mass ratio of 5:3: 1;
the auxiliary material filling is selected from one or more of maltodextrin, citric acid, fruit powder and sugar alcohol.
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| RU2679421C1 (en) * | 2018-02-12 | 2019-02-08 | Государственное научное учреждение Всероссийский научно-исследовательский ветеринарный институт патологии, фармакологии и терапии Российской академии сельскохозяйственных наук (ГНУ ВНИВИПФиТ Россельхозакадемии) | Method of prevention of inflammatory pathology in reproductive system of sows |
| CN108902981A (en) * | 2018-04-25 | 2018-11-30 | 深圳市多微生保健食品有限公司 | A kind of probiotic composition and probiotics solid beverage |
| CN110917174A (en) * | 2018-09-20 | 2020-03-27 | 广东怡和科洁科技有限公司 | Oral probiotic capsule for regulating balance of female vaginal flora and preparation method thereof |
| CN110771766A (en) * | 2019-11-13 | 2020-02-11 | 上海英库商务咨询有限公司 | Probiotic health-care beverage for treating child intestinal colic and diarrhea and repairing intestinal tract |
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| CN117165465A (en) * | 2021-08-24 | 2023-12-05 | 上海市第一人民医院 | Lactobacillus plantarum strain for improving intestinal barrier function and application thereof |
| CN114921389A (en) * | 2022-07-20 | 2022-08-19 | 广东益可维生物技术有限公司 | Probiotic composition with female intestinal private part nursing and mammary gland anti-inflammatory effects and application thereof |
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