A composition containing probiotics for regulating oral flora and oral micro-effervescent tablet and application thereof

Abstract

The present invention belongs to the field of microbial technology, and specifically relates to a composition containing probiotics for regulating oral flora, an oral micro-effervescent tablet and an application thereof, wherein the composition comprises active ingredients Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin. Experiments show that the composition of the present invention can significantly reduce the content of TNF-γ, IgE and IL-17 in the serum of asthmatic rats. This shows that after the combination of Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin, there is a significant synergistic effect in the effect of preventing and treating asthma, and a technical effect that is difficult for technicians in the field to foresee is obtained.

Description

Technical Field

[0001] The invention relates to the technical field of microorganisms, and in particular to a composition containing probiotics for regulating oral flora, an oral micro-effervescent tablet and applications thereof. Background Art

[0002] Probiotics are a type of active microorganisms that are beneficial to the host by colonizing in the human body and changing the composition of the flora in a certain part of the host. They promote nutrient absorption by regulating the host's mucosal and systemic immune functions or regulating the balance of intestinal flora, thereby producing benefits to the host's health.

[0003] The oral cavity is the second largest habitat for microbial colonies after the intestines. The oral cavity, which is fully connected to the air, also has a large number of anaerobic bacteria. The oral flora can change the balance between local and systemic health and disease; there are bacteria, fungi, archaea, protozoa and viruses in the oral cavity, and the interactions between various species and the host have an impact on the health and disease of the host. The oral microecology is closely related to diseases such as obesity, liver fibrosis, inflammatory bowel disease, polycystic ovary syndrome, adverse pregnancy, asthma, atherosclerosis, and pancreatic cancer.

[0004] Bronchial asthma (abbreviated as asthma) is a chronic airway disease, which is mainly manifested by reversible airway obstruction, airway hyperresponsiveness, eosinophils and CD4+T helper (Th) type 2 inflammatory infiltration and airway remodeling. Symptoms include wheezing, dyspnea, chest tightness and cough. It is currently believed that the pathogenesis of asthma is the result of the combined effects of genetics and the environment, mainly including genetic mechanisms, airway inflammation-immune mechanisms and neural regulation mechanisms. In recent years, its prevalence has been increasing year by year worldwide, bringing heavy economic and psychological burdens to patients' families.

[0005] At present, the main methods of asthma treatment are anti-inflammatory and bronchodilator therapy. The more effective drugs for treating asthma are β2-receptor agonists, leukotriene modifiers, theophylline, glucocorticoids, antihistamines and anticholinergic drugs. These drugs can control the symptoms of asthma, but they cannot cure asthma. In addition, these drugs take a long time to treat and have many side effects. A large number of patients cannot tolerate them and have poor compliance. It is of great significance to find effective prevention and treatment plans.

[0006] A large number of epidemiological studies have shown that exposure to beneficial microorganisms can reduce the occurrence and development of asthma and can be used to relieve asthma. These beneficial microorganisms, live or dead, mainly regulate the human immune function through the oral cavity, respiratory tract and digestive tract, inhibiting the occurrence, development and development of asthma. A large number of studies have found that probiotics have immunomodulatory and anti-infective activities and can be used to relieve asthma. However, the effect of using probiotics alone to relieve asthma is limited. Therefore, how to further improve the effect of probiotics in relieving asthma is still a technical problem that needs to be solved by technicians in this field.

[0007] Studies have shown that Lactobacillus salivarius HA-118 derived from the human body is beneficial to oral health and balances the oral microecology. Lactobacillus salivarius HA-118 can metabolize to produce antibacterial components H 2 O 2 , reducing volatile sulfides in the oral cavity, and has a significant effect on oral odor (bad breath). At the same time, HA-118 has a good inhibitory effect on oral pathogens-Streptococcus mutans. HA-118 has strong oral persistence against periodontal pathogens such as Streptococcus mutans, Streptococcus sanguinis, and Pyrromonas. Lactobacillus helveticus Lafti L10 derived from dairy products can also metabolize to produce antibacterial components H 2 O 2 , significantly reducing volatile sulfides in the oral cavity, and has a significant effect on oral odor (bad breath). Lafti L10 has a good inhibitory effect on Candida albicans and can prevent mucosal infections. Lafti L10 also has the effect of regulating immunity, which can reduce cold symptoms and drug use. Bifidobacterium longum R0175 isolated from the human body can act on the "brain-gut axis" by protecting the intestinal barrier, reducing the production of proinflammatory factors, promoting the synthesis of neurotransmitters, regulating the vagus nerve, etc., and has good anti-depressant and anti-anxiety effects in clinical experiments, thereby having a positive impact on mental health. Lactobacillus rhamnosus R0011 isolated from dairy products has good adhesion in the intestine, can reduce the colonization of Helicobacter pylori and the inflammation caused by it, and can relieve gastric mucosal inflammatory reactions by reducing the colonization of Helicobacter pylori. Combinations of probiotics often have higher clinical efficacy than any single probiotic preparation. The probiotic combination of Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175 and Lactobacillus rhamnosus R0011 has also been shown in multiple clinical studies to control the growth of Streptococcus mutans, maintain the pH value of healthy saliva, help reduce or slow the formation of dental plaque, and promote gum health. Compared with widely used mouthwashes containing chlorhexidine or sodium fluoride, this probiotic combination provides similar or better effects. And this specific probiotic combination can regulate the healthy balance of oral microbiota, support the healthy level of Candida in the oral microbiome, and maintain healthy breath.

[0008] Fucoxanthin is the most abundant compound among carotenoids, accounting for more than 10% of the total production of about 700 naturally occurring carotenoids. It is widely found in various algae, marine phytoplankton, aquatic shellfish and other animals and plants. Studies have shown that fucoxanthin has a highly unique structure, containing both epoxy bonds and hydroxyl groups in the polyene chain, as well as allylic bonds and conjugated carbonyl groups. These structures give fucoxanthin a variety of biological activities such as antioxidant, anti-cancer, anti-inflammatory, neuronal cell protection, and weight loss, and can be widely used in pharmaceuticals and skin care and beauty products.

[0009] At present, there is no research on the combined use of Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin in the prevention and treatment of asthma. Summary of the invention

[0010] The purpose of the present invention is to provide a composition containing probiotics for regulating oral flora with synergistic effects and an oral micro-effervescent tablet and application thereof, so as to provide a new idea for preventing and treating asthma safely, effectively, conveniently and economically.

[0011] Specifically, the present invention is realized through the following technical solutions.

[0012] In a first aspect, the present invention provides a composition containing probiotics for regulating oral flora, the composition comprising active ingredients Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin.

[0013] In a preferred embodiment of the present invention, the composition comprises, by weight, 2.5-15 parts of Lactobacillus salivarius HA-118, 2.5-15 parts of Lactobacillus helveticus Lafti L10, 5-30 parts of Bifidobacterium longum R0175, 5-30 parts of Lactobacillus rhamnosus R0011 and 10-60 parts of fucoxanthin.

[0014] In a preferred embodiment of the present invention, the composition comprises, by weight, 5-10 parts of Lactobacillus salivarius HA-118, 5-10 parts of Lactobacillus helveticus Lafti L10, 10-20 parts of Bifidobacterium longum R0175, 10-20 parts of Lactobacillus rhamnosus R0011 and 30-50 parts of fucoxanthin.

[0015] In a preferred embodiment of the present invention, in the above composition, the viable count of Lactobacillus salivarius HA-118 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0016] In a preferred embodiment of the present invention, in the above composition, the viable count of Lactobacillus helveticus Lafti L10 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0017] In a preferred embodiment of the present invention, in the above composition, the viable count of Bifidobacterium longum R0175 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0018] In a preferred embodiment of the present invention, in the above composition, the viable count of Lactobacillus rhamnosus R0011 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0019] In a preferred embodiment of the present invention, in the above composition, the Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, and Lactobacillus rhamnosus R0011 are freeze-dried bacterial powders.

[0020] In a second aspect, the present invention provides use of the composition described in the first aspect in preparing a medicine for preventing and treating asthma.

[0021] In a preferred embodiment of the present invention, in the above-mentioned medicine, the viable count of Lactobacillus salivarius HA-118 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0022] In a preferred embodiment of the present invention, in the above-mentioned medicine, the viable count of Lactobacillus helveticus Lafti L10 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0023] In a preferred embodiment of the present invention, in the above-mentioned medicine, the viable count of Bifidobacterium longum R0175 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0024] In a preferred embodiment of the present invention, in the above-mentioned medicine, the viable count of Lactobacillus rhamnosus R0011 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0025] In a preferred embodiment of the present invention, in the above-mentioned medicine, the Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, and Lactobacillus rhamnosus R0011 are freeze-dried bacterial powders.

[0026] In a preferred embodiment of the present invention, the medicine for preventing and treating asthma may further include a pharmaceutically acceptable carrier, and the pharmaceutically acceptable carrier is selected from one or more of fillers, disintegrants, lubricants, binders, wetting agents, suspending agents, surfactants, flavoring agents, sweeteners, solvents, acid sources, alkali sources, pigments, etc.

[0027] In a preferred embodiment of the present invention, the filler includes but is not limited to corn starch, lactose, pregelatinized starch, microcrystalline cellulose, maltodextrin, etc.; the disintegrant includes but is not limited to starch effervescent mixture, i.e. sodium bicarbonate and citric acid, tartaric acid, low-substituted hydroxypropyl cellulose, etc.; the lubricant includes but is not limited to talc, sodium lauryl sulfate, sodium oleate, stearic acid, poloxamer, magnesium stearate, etc.; the binder includes but is not limited to hydroxypropyl methylcellulose, syrup, starch slurry, sodium carboxymethyl cellulose, etc.; the suspending agent includes but is not limited to polysaccharides such as agar, acacia gum, cellulose ether, alginic acid and carboxymethyl chitosan, etc.; the acid source includes but is not limited to citric acid, tartaric acid, malic acid and citric acid, etc.; the alkali source includes but is not limited to sodium bicarbonate, sodium carbonate, etc.

[0028] In a third aspect, the present invention provides an oral micro-effervescent tablet for preventing and treating asthma, wherein the oral micro-effervescent tablet comprises the composition described in the first aspect.

[0029] In a preferred embodiment of the present invention, the above-mentioned oral micro-effervescent tablet further comprises auxiliary materials such as an acid source, an alkali source, a flavoring agent, a sweetener, an essence, and magnesium stearate.

[0030] In a preferred embodiment of the present invention, in the above-mentioned oral micro-effervescent tablet, the viable count of Lactobacillus salivarius HA-118 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0031] In a preferred embodiment of the present invention, in the above-mentioned oral micro-effervescent tablet, the viable count of Lactobacillus helveticus LaftiL10 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0032] In a preferred embodiment of the present invention, in the above-mentioned oral micro-effervescent tablet, the viable count of Bifidobacterium longum R0175 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0033] In a preferred embodiment of the present invention, in the above-mentioned oral micro-effervescent tablet, the viable count of Lactobacillus rhamnosus R0011 is 1×10 8 cfu / g-1×10 10 cfu / g.

[0034] In a preferred embodiment of the present invention, in the above-mentioned oral micro-effervescent tablet, the Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, and Lactobacillus rhamnosus R0011 are freeze-dried bacterial powders.

[0035] In the present invention, the above-mentioned composition and oral micro-effervescent tablet can be prepared according to conventional processes in the field of pharmaceutical preparations.

[0036] Compared with the prior art, the present invention has the following beneficial effects:

[0037] The present invention creatively develops a composition containing probiotics for regulating oral flora. By combining Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin in a specific dosage ratio, the content of TNF-γ, IgE and IL-17 in the serum of asthmatic rats can be significantly reduced, and the prevention and treatment effect of asthma can be greatly improved. The effect is significantly better than the same dose of Lactobacillus salivarius HA-118 and Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin used alone, and significantly better than the same dose of four probiotics used in combination. This shows that the combination of Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin has a significant synergistic effect in the prevention and treatment of asthma, and obtains technical effects that are difficult for technicians in the field to foresee. Moreover, the composition of the present invention has no toxic side effects and is easy to use. It can be used to prepare related products for preventing and treating asthma and has broad application prospects. DETAILED DESCRIPTION

[0038] The present invention will be further described with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0039] If no specific techniques or conditions are specified in the examples, the techniques or conditions described in the literature in the field or the product instructions are used. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0040] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0041] Example:

[0042] This example explores the effect of the probiotic composition of the present invention on the levels of TNF-γ, IgE, and IL-17 in the serum of an asthma rat model:

[0043] 1. Main experimental materials

[0044] (1) Experimental animals

[0045] 90 healthy male clean-grade SD rats weighing 180-200 g were selected and provided by the Animal Experiment Center of Southern Medical University. They were kept for 3 days before the experiment to adapt to the environment.

[0046] (2) Experimental Materials

[0047] Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011, fucoxanthin, dexamethasone, rat TNF-γ ELISA kit, rat IgE ELISA kit and rat IL-17 ELISA kit. The above strains were made into freeze-dried bacterial powders respectively.

[0048] 2. Experimental Methods

[0049] (1) Establishment of asthma rat model

[0050] After 3 days of adaptive feeding, the rats except the blank group were injected with 2 ml of 10% egg protein + 10% aluminum hydroxide saline solution on the 1st and 8th days of the experiment, and 0.5 ml was injected subcutaneously at each point on both sides of the waist to sensitize the rats. From the 15th day, 6% egg protein saline was used for aerosol inhalation to stimulate asthma in rats, with an aerosol flow rate of 10 ml / min, 20 rain each time, once every other day, for a total of 4 weeks. The signs of successful stimulation were cyanosis of the lips, abdominal muscle spasms, accelerated breathing, nodding breathing, and unstable standing in rats. At the corresponding time, the blank group was injected with saline instead of egg protein for intraperitoneal injection and aerosol inhalation.

[0051] (2) Rats grouping

[0052] The dosage conversion method for rats is based on the daily probiotic consumption of 9 billion cfu per person. The average human body weight is 60 kg, and the dosage for each kilogram of body weight is 150 million cfu of probiotics. The rat dosage is 8 times that of humans, that is, 1.2 billion cfu / kg.

[0053] A total of 90 rats were randomly divided into 9 groups, with 10 rats in each group, namely, blank group, model group, dexamethasone group, Lactobacillus salivarius HA-118 + Lactobacillus helveticus Lafti L10 group (HA-118 + Lafti L10 group), Bifidobacterium longum R0175 group (R0175 group), Lactobacillus rhamnosus R0011 group (R0011 group), fucoxanthin group, HA-118 + Lafti L10 + R0175 + R0011 group, and HA-118 + Lafti L10 + R0175 + R0011 + fucoxanthin group. The model group and the blank group were gavaged with an equal amount of saline half an hour before the challenge; the dexamethasone group was given dexamethasone 6 mg / kg half an hour before the challenge; the HA-118+LaftiL10 group was given 600 million cfu / kgHA-118+600 million cfu / kgLafti L10 half an hour before the challenge; the R0175 group was given 1.2 billion cfu / kgR0175 half an hour before the challenge; the R0011 group was given 1.2 billion cfu / kgR0011 half an hour before the challenge; the HA-118+LaftiL10+R0175+R0011 group was given 200 million cfu / kgHA-118+200 million cfu / kgLafti L10+400 million cfu / kgR0175+400 million cfu / kgR0011 half an hour before the challenge; the fucoxanthin group was given 80 mg / kg fucoxanthin half an hour before the challenge; The L10+R0175+R0011+fucoxanthin group was given 200 million cfu / kg HA-118+200 million cfu / kg Lafti L10+400 million cfu / kg R0175+400 million cfu / kg R0011+80 mg / kg fucoxanthin half an hour before the challenge. Each group was given the corresponding substances by gavage every day from the first asthma challenge until before sacrifice for 4 consecutive weeks.

[0054] (3) Specimen collection

[0055] Blood samples were extracted 2 hours after the last administration of the corresponding substances. Blood was collected from the jugular vein, left to stand for 8 hours, and then centrifuged at 4000 r / min for 10 minutes. The upper layer of serum was collected and kept at low temperature for later use.

[0056] (4) Detection of related cytokine expression in serum

[0057] The expressions of related cytokines TNF-γ, IgE, and IL-17 were detected by ELISA.

[0058] (5) Statistical methods

[0059] The results were analyzed using SPSS 19.0 statistical software. One-way analysis of variance was used for comparison among multiple groups, and t-test was used for comparison between two groups. P<0.05 indicated that the difference was statistically significant, and P<0.01 indicated that the difference was statistically significant.

[0060] 3. Experimental Results

[0061] The expressions of related cytokines TNF-γ, IgE, and IL-17 in serum are shown in Table 1.

[0062] As shown in Table 1, compared with the blank group, the levels of TNF-γ, IgE, and IL-17 in the serum of asthmatic rats in the model group were significantly increased (P<0.01); compared with the model group, the levels of TNF-γ, IgE, and IL-17 in the serum of asthmatic rats in the HA-118+Lafti L10+R0175+R0011+fucoxanthin group (P<0.01) and dexamethasone group (P<0.01) were statistically significantly decreased. From these results, it can be seen that when HA-118, Lafti L10, R0175, R0011 and fucoxanthin are used in combination, the combined group has a significantly better effect on reducing the levels of TNF-γ, IgE and IL-17 in the serum of asthmatic rats than the HA-118+Lafti L10, R0175, R0011 and fucoxanthin alone groups, and the effect of the combined group is also significantly better than the probiotic combination group. This shows that the combination of HA-118, Lafti L10, R0175, R0011 and fucoxanthin has a significant synergistic effect in reducing the levels of TNF-γ, IgE and IL-17 in the serum of asthmatic rats.

[0063] Table 1: TNF-γ, IgE, IL-17 levels in the serum of rats in each group (x±s, n=10)

[0064]

[0065] 4. Experimental Conclusion

[0066] Asthma is a chronic airway disease involving a variety of inflammatory cells such as T lymphocytes, eosinophils, neutrophils and airway epithelial cells. Therefore, the detection of inflammatory factors TNF-γ, IgE, and IL-17 levels are directly related to asthma, and reducing the above indicators can reflect the preventive and therapeutic effects on asthma. The experimental results showed that when Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin were used in combination, the combined group significantly reduced the levels of TNF-γ, IgE, and IL-17 in the serum of asthmatic rats than the HA-118+Lafti L10, R0175, R0011, fucoxanthin alone group and the probiotic combination group. This shows that the combined group has a significantly better preventive and therapeutic effect on asthma than the same dose of HA-118+Lafti L10, R0175, R0011 and fucoxanthin alone group and the probiotic combination group. This shows that the combined use of Lactobacillus salivarius HA-118, Lactobacillus helveticus Lafti L10, Bifidobacterium longum R0175, Lactobacillus rhamnosus R0011 and fucoxanthin has a significant synergistic effect on the preventive and therapeutic effect of asthma. Therefore, the composition containing probiotics of the present invention can be used to prepare related products for the prevention and treatment of asthma, and has a wide range of application prospects.

[0067] Obviously, the above embodiments are only examples for clearly explaining the present invention, and are not intended to limit the implementation methods of the present invention. Those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Use of a composition containing probiotics for regulating oral flora in the preparation of a medicine for preventing and treating asthma, characterized in that: The composition is prepared from 5-10 parts of Lactobacillus salivarius HA-118, 5-10 parts of Lactobacillus helveticus Lafti L10, 10-20 parts of Bifidobacterium longum R0175, 10-20 parts of Lactobacillus rhamnosus R0011 and 30-50 parts of fucoxanthin by weight.

2. The use according to claim 1, characterized in that: In the drug, the number of viable bacteria of Lactobacillus salivarius HA-118 is 1×10 8 cfu / g-1×10 10 cfu / g; the viable count of Lactobacillus helveticus Lafti L10 is 1×10 8 cfu / g-1×10 10 cfu / g; the viable count of Bifidobacterium longum R0175 is 1×10 8 cfu / g-1×10 10 cfu / g; the viable count of Lactobacillus rhamnosus R0011 was 1×10 8 cfu / g-1×10 10 cfu / g.

3. The use according to claim 1, characterized in that: The medicine is an oral micro-effervescent tablet, and the oral micro-effervescent tablet comprises the composition described in claim 1.

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