阅读说明：本技术 用于治疗特应性皮炎的微生物复合制剂 (Microbial complex preparation for treating atopic dermatitis ) 是由 刘卫 于 2021-06-02 设计创作，主要内容包括：本发明属于皮肤治疗制剂技术领域,公开了用于治疗特应性皮炎的微生物复合制剂,本发明提供了用于治疗特应性皮炎的微生物复合制剂,包括植物乳杆菌NumRes8、鼠李糖乳杆菌HN001、木耳多糖、长双歧杆菌BB536、大豆皂苷、丁酸梭菌JC10390；该微生物复合制剂通过调节患者免疫反应来改善患者的生活质量,有效缓解特应性皮炎患者的症状,且里面涉及的原料容易获得,成本低,对人体无副作用,为特应性皮炎的治疗和干预提供新的方案。(The invention belongs to the technical field of skin treatment preparations, and discloses a microbial compound preparation for treating atopic dermatitis, which comprises lactobacillus plantarum NumRes8, lactobacillus rhamnosus HN001, agaric polysaccharide, bifidobacterium longum BB536, soyasaponin, clostridium butyricum JC 10390; the microbial compound preparation can improve the quality of life of patients by regulating the immune response of the patients, effectively relieve the symptoms of atopic dermatitis patients, and the raw materials involved in the preparation are easy to obtain, low in cost and free of side effects on human bodies, thereby providing a new scheme for the treatment and intervention of atopic dermatitis.)

1. The microbial compound preparation for treating atopic dermatitis is characterized by comprising lactobacillus plantarum NumRes8, lactobacillus rhamnosus HN001, agaric polysaccharide, bifidobacterium longum BB536, soyasaponin and clostridium butyricum JC 10390.

2. The complex microbial preparation for the treatment of atopic dermatitis according to claim 1, which is composed of the following components in parts by weight:

3. the microbial complex preparation for treating atopic dermatitis according to claim 2, wherein the amount of inactivated bacteria of Lactobacillus plantarum NumRes8 is 3.1 to 3.8 x 10¹²Per gram.

4. The microbial complex preparation for treating atopic dermatitis according to claim 2, wherein the inactivated bacterial amount of Lactobacillus rhamnosus HN001 is 2.5 to 3.1 x 10¹²Per gram.

5. The microbial complex preparation for treating atopic dermatitis according to claim 2, wherein the inactivated amount of Bifidobacterium longum BB536 is 3.5 to 4.8 x 10¹¹Per gram.

6. The microbial complex preparation for treating atopic dermatitis according to claim 2, wherein the inactivated bacterial amount of Clostridium butyricum JC10390 is 1.5 to 2.8 x 10⁹Per gram.

7. The microbial composite preparation according to claim 2, wherein the preparation is in the form of a liquid, a gas or a solid.

8. Use of the microbial complex according to any one of claims 1 to 7 in the manufacture of a medicament for the treatment and/or alleviation and/or prevention of atopic dermatitis.

Technical Field

The invention relates to the technical field of skin treatment preparations, in particular to a microbial compound preparation for treating atopic dermatitis.

Background

Atopic dermatitis is a chronic, recurrent, inflammatory skin disease that is clinically manifested primarily as recurrent chronic eczematous rashes with marked dryness and itching of the skin. Patients are considered to be a systemic disease because they often incorporate other atopic diseases such as allergic rhinitis and asthma.

Atopic dermatitis is characterized by a distinct "atopic" trait seen in patients or their families: firstly, the familial tendency of asthma, allergic rhinitis and eczema is easy to suffer; ② allergy to foreign proteins; ③ high IgE in serum; fourthly, eosinophilia in blood. Typical atopic dermatitis has specific clinical manifestations of eczema and four of the above mentioned features. Also known as atopic dermatitis, atopic eczema, Besnier prurigo in body or atopic eczema. The clinical stage is divided into three stages: acute or subacute eczematous in infancy; the childhood and adolescence are subacute or chronic eczema.

The current therapeutic drugs for atopic dermatitis are classified into the following categories: 1. glucocorticoids, such as cyclosporine, azathioprine; it has the effects of regulating synthesis and metabolism of saccharide, fat and protein in vivo, and most importantly, can exert the effects of suppressing immune response and resisting allergy in allergic diseases; 2. the immunosuppressant can be used for regulating and inhibiting the activity of calcineurin in the process of immune signal transduction, so as to inhibit the gene transcription necessary for activating the nuclear factor of T lymphocyte, further inhibit the induced expression of cell factors such as IL-3, IL-4, GM-CSF and the like in immune reaction, and finally play the role of immune anti-inflammation; 3. traditional Chinese medicines such as Petuqingxin formula are also used for treating atopic dermatitis; 4. biological agents such as recombinant human interferon gamma (IFN-gamma), IL-4 receptor antagonists, and the like. The above-mentioned therapeutic drugs may have side effects, for example, the skin atrophy and aggravation of infection due to long-term use of glucocorticoid, and the other unknown effects in vivo due to use of immunosuppressant and biological agent

To date, atopic dermatitis is not currently curable and can severely affect quality of life, but effective and normative treatment can alleviate symptoms, reducing recurrence; therefore, the method has positive significance for daily treatment management and intervention of atopic dermatitis.

Disclosure of Invention

The present invention has been made to overcome the above-mentioned problems occurring in the prior art, and firstly provides a microbial preparation for treating atopic dermatitis.

The purpose of the invention is realized by the following technical scheme:

a compound microorganism preparation for treating atopic dermatitis comprises Lactobacillus plantarum NumRes8, Lactobacillus rhamnosus HN001, Auricularia polysaccharide, Bifidobacterium longum BB536, soyasaponin, Clostridium butyricum JC 10390.

Atopic dermatitis belongs to the category of allergic diseases, and the mechanism of investigation on atopic dermatitis has been found in recent years: more than 80% of atopic dermatitis patients are accompanied by an increase in the level of specific IgE and, further, the reason behind this phenomenon is analyzed that atopic dermatitis occurs involving multiple links of immune response, uptake and presentation of allergens by skin antigen presenting cells such as langerhans cells, dendritic cells, excessive recruitment of mast cells and eosinophils, leading to excessive production of inflammatory mediators and cytokines by keratinocytes, and a dysbalance Th17/Th22, but the central mechanism is, overall, an immunological abnormality dominated by a Th2 type reaction; in this process, the production of some immunoinflammatory factors such as IL-4, IL-5 and IL-13 exacerbates Th2 type responses, leading to impaired skin barrier; in addition, due to the abundant expression of Th2 type cytokines, CD4+ T cells interact with B cells under the action of the cytokine IL-4 and are plasma cells that secrete specific IgE, thus leading to elevated IgE levels.

The existing research shows that the intake of probiotics is helpful for relieving or preventing allergic diseases, but no exact conclusion is made at present on the way that the specific probiotics play a role in preventing and treating the allergic diseases, and in addition, the participation of the probiotics in regulating the immune system of a host body has a great relationship with the genome of the probiotics and the genome of the host body, the environment and the dosage of the probiotics, and the host individual difference exists, so that for the specific type of allergic diseases, a great deal of research needs to be carried out, and the types of specific microorganisms for treating, assisting in treating and relieving the allergic diseases are obtained through statistics.

In the microbial compound preparation, the lactobacillus plantarum NumRes8 can reduce the amounts of IL-4 and IL-5 in animal blood, so that the allergic reaction degree of skin is relieved; the Auricularia polysaccharide can inhibit release of inflammatory cytokine such as IL-4, and inhibit IgE antibody production, thereby relieving anaphylaxis; bifidobacterium longum BB536 can increase IFN-gamma expression level of skin tissue, and inhibit IL-4 and IL-5 secretion; lactobacillus rhamnosus HN001 can reduce IgE level in serum; the soyasaponin is helpful for enhancing the expression of IFN-gamma of skin tissues so as to resist the caused skin itch; clostridium butyricum JC10390 helps to increase the expression of IL-10 and IFN-gamma in skin tissues, thereby stimulating Th1 type immune response and regulating Th1/Th2 immune balance.

Preferably, the microbial composite preparation for treating atopic dermatitis comprises the following components in parts by weight:

preferably, the amount of the inactivated lactobacillus plantarum NumRes8 in the microbial composite preparation for treating atopic dermatitis is 3.1-3.8 x 10¹²Per gram.

Preferably, the inactivated bacterium amount of lactobacillus rhamnosus HN001 in the microbial composite preparation for treating atopic dermatitis is 2.5-3.1 × 10¹²Per gram.

Preferably, the amount of the inactivated bacteria of Bifidobacterium longum BB536 in the microbial composite preparation for treating atopic dermatitis is 3.5-4.8 x 10¹¹Per gram.

Preferably, the inactivated bacterial amount of clostridium butyricum JC10390 in the microbial compound preparation for treating atopic dermatitis is 1.5-2.8 x 10⁹Per gram.

Preferably, the formulation of the microbial composite preparation for treating atopic dermatitis is a liquid formulation, a gaseous formulation and a solid formulation.

The above-mentioned microbial complex preparation of the present invention can be administered orally or by mucosal administration, and thus can be used for the treatment and/or adjuvant treatment and/or prevention of atopic dermatitis.

Therefore, the invention also provides the application of the microbial compound preparation in preparing a medicament for treating and/or relieving and/or preventing atopic dermatitis.

Compared with the prior art, the invention has the following beneficial effects:

the invention provides a microbial compound preparation for treating atopic dermatitis, which comprises lactobacillus plantarum NumRes8, lactobacillus rhamnosus HN001, agaric polysaccharide, bifidobacterium longum BB536, soyasaponin, clostridium butyricum JC 10390; in the microbial compound preparation, the lactobacillus plantarum NumRes8 can reduce the amounts of IL-4 and IL-5 in animal blood, thereby relieving the allergic reaction degree of skin; the Auricularia polysaccharide can inhibit release of inflammatory cytokine such as IL-4, and inhibit IgE antibody production, thereby relieving anaphylaxis; bifidobacterium longum BB536 can increase IFN-gamma expression level of skin tissue, and inhibit IL-4 and IL-5 secretion; lactobacillus rhamnosus HN001 can reduce IgE level in serum; the soyasaponin is helpful for enhancing the expression of IFN-gamma of skin tissues so as to resist the caused skin itch; clostridium butyricum JC10390 helps to increase the expression of IL-10 and IFN-gamma in skin tissues, thereby stimulating Th1 type immune response and regulating Th1/Th2 immune balance.

The microbial compound preparation can improve the quality of life of patients by regulating the immune response of the patients, effectively relieve the symptoms of atopic dermatitis patients, and the raw materials involved in the preparation are easy to obtain, low in cost and free of side effects on human bodies, thereby providing a new scheme for the treatment and intervention of atopic dermatitis.

Detailed Description

The following further describes the embodiments of the present invention. It should be noted that the description of the embodiments is provided to help understanding of the present invention, but the present invention is not limited thereto. In addition, the technical features involved in the embodiments of the present invention described below may be combined with each other as long as they do not conflict with each other.

The test methods used in the following experimental examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.

Example 1

The microbial compound preparation for treating atopic dermatitis consists of the following components in parts by weight: 2 parts of lactobacillus plantarum NumRes8, 1 part of lactobacillus rhamnosus HN001, 2 parts of agaric polysaccharide, 3 parts of bifidobacterium longum BB536, 1 part of soyasaponin and 1 part of clostridium butyricum JC 10390.

Wherein the amount of the inactivated lactobacillus plantarum NumRes8 was 3.5X 10¹²Per gram; the inactivated bacterial amount of Lactobacillus rhamnosus HN001 is 2.5 × 10¹²Per gram; the inactivated bacterial amount of Bifidobacterium longum BB536 is 3.5X 10¹¹Per gram; the inactivated bacterial amount of Clostridium butyricum JC10390 is 2.0 × 10⁹Per gram.

Example 2

The microbial compound preparation for treating atopic dermatitis consists of the following components in parts by weight: 3 parts of lactobacillus plantarum NumRes8, 2 parts of lactobacillus rhamnosus HN001, 1 part of agaric polysaccharide, 2 parts of bifidobacterium longum BB536, 1 part of soyasaponin and 1 part of clostridium butyricum JC 10390.

Wherein the amount of the inactivated lactobacillus plantarum NumRes8 was 3.1X 10¹²Per gram; the inactivated bacterial amount of Lactobacillus rhamnosus HN001 is 2.8 × 10¹²Per gram; the inactivated bacterial amount of Bifidobacterium longum BB536 is 4.0X 10¹¹Per gram; the inactivated bacterial amount of Clostridium butyricum JC10390 is 1.5X 10⁹Per gram.

Example 3

The microbial compound preparation for treating atopic dermatitis consists of the following components in parts by weight: 4 parts of lactobacillus plantarum NumRes8, 1 part of lactobacillus rhamnosus HN001, 1 part of agaric polysaccharide, 3 parts of bifidobacterium longum BB536, 0.5 part of soyasaponin and 0.5 part of clostridium butyricum JC 10390.

Wherein the amount of the inactivated lactobacillus plantarum NumRes8 was 3.1X 10¹²Per gram; the inactivated bacterial amount of Lactobacillus rhamnosus HN001 is 3.1 × 10¹²Per gram; the inactivated bacterial amount of Bifidobacterium longum BB536 is 4.0X 10¹¹Per gram; the inactivated bacterial amount of Clostridium butyricum JC10390 is 2.8 multiplied by 10⁹Per gram.

Example 4

The microbial compound preparation for treating atopic dermatitis consists of the following components in parts by weight: 2 parts of lactobacillus plantarum NumRes8, 1 part of lactobacillus rhamnosus HN001, 1 part of agaric polysaccharide, 4 parts of bifidobacterium longum BB536, 1 part of soyasaponin and 1 part of clostridium butyricum JC 10390.

Wherein the amount of the inactivated lactobacillus plantarum NumRes8 was 3.8X 10¹²Per gram; the inactivated bacterial amount of Lactobacillus rhamnosus HN001 is 3.0 × 10¹²Per gram; the inactivated bacterial amount of Bifidobacterium longum BB536 is 3.5X 10¹¹Per gram; the inactivated bacterial amount of Clostridium butyricum JC10390 is 2.5X 10⁹Per gram.

Example 5

The microbial compound preparation for treating atopic dermatitis consists of the following components in parts by weight: 3 parts of lactobacillus plantarum NumRes8, 2 parts of lactobacillus rhamnosus HN001, 2 parts of agaric polysaccharide, 2 parts of bifidobacterium longum BB536, 0.5 part of soyasaponin and 0.5 part of clostridium butyricum JC 10390.

Wherein the amount of the inactivated lactobacillus plantarum NumRes8 was 3.5X 10¹²Per gram; the inactivated bacterial amount of Lactobacillus rhamnosus HN001 is 3.1 × 10¹²Per gram; the inactivated bacterial amount of Bifidobacterium longum BB536 is 4.8 × 10¹¹Per gram; the inactivated bacterial amount of Clostridium butyricum JC10390 is 2.8 multiplied by 10⁹Per gram.

Comparative example 1

A microbial composite preparation for the treatment of atopic dermatitis, which had substantially the same formulation as in example 2, except that in this comparative example, Lactobacillus plantarum L-137 was used as the strain of Lactobacillus plantarum.

Comparative example 2

A microbial complex preparation for the treatment of atopic dermatitis, having substantially the same formulation as in example 2, except that in this comparative example, Bifidobacterium longum strain used was Bifidobacterium longum E4.

Comparative example 3

A complex microbial preparation for the treatment of atopic dermatitis having substantially the same formulation as in example 2, except that in this comparative example, the strain of Lactobacillus rhamnosus LGG was used.

Comparative example 4

A microbial complex preparation for the treatment of atopic dermatitis having substantially the same formulation as in example 2, except that the fungus polysaccharide is not contained in the present comparative example.

Comparative example 5

The formulation of the microbial composite preparation for treating atopic dermatitis was substantially the same as in example 2, except that in this comparative example, soyasaponin was not contained.

Comparative example 6

A microbial composite preparation for the treatment of atopic dermatitis, having substantially the same formulation as in example 2, except that in this comparative example, the auricularia auricula polysaccharide was replaced with phellinus linteus polysaccharide.

Examples of the experiments

First, animal experiment

1. Experimental animals: SPF6 week old male mice.

2. Experiment design: the experimental mice are normally fed and raised for 4 weeks, the first week is the adaptation period of the mice, the second week begins, the intervention group can be intragastrically filled (0.1mL/kg body weight) according to the actual formula, the blank group, the model group can be intragastrically filled with the same amount of sterile physiological saline, and the positive drug group can be intragastrically filled with the same amount of Keto drug solution (1mg/kg body weight) until the experiment is finished.

The mouse modeling experiment refers to the prior art and comprises the following steps: the skin hairs on the back of the mice were shaved off by a shaver before molding, and the size was 2.5cm long by 1.5cm wide. A base solution is prepared by acetone and olive oil in a ratio of 4:1, then DNFB and the base solution are used for preparing 0.5% and 0.2% of DNFB sensitizing drugs, modeling is carried out at the beginning of 3 weeks, at the 1 st d of modeling, 25 mu L of DNFB with the concentration of 0.5% is used for smearing sensitization on the back skin and the right ear of a mouse, then 20 mu L of DNFB solution with the concentration of 0.2% is used for smearing atopic dermatitis on the back skin and the right ear of the mouse at the 5 th, 8 th, 11 th and 14 th d, and the same amount of base solution is smeared on blank mice. After the model building is finished, the mice are anesthetized by isoflurane and killed, the blood of the mice is centrifuged at 5600r/min for 15min at room temperature to collect serum, and the back skin of the mice is taken down and homogenized for detecting the immune inflammatory factors.

3. Mouse ear swelling degree determination

After the mice died, the thickness of the middle part of the right ear of the mice was measured by a specially trained person with an electronic vernier caliper and recorded.

4. Determination of immunoinflammatory factors: the procedure was performed according to the existing kit.

Second, experimental results

1. Effect of degree of ear swelling in atopic dermatitis mice

The experimental results are shown in table 1, and it can be seen from table 1 that the embodiments of the technical scheme adopted by the present invention (examples 1 to 5) all have a relatively significant effect of relieving the swelling degree of the mouse ear, compared with the comparative examples 1 to 3, the lactobacillus plantarum, lactobacillus rhamnosus, clostridium butyricum and bifidobacterium longum adopted by the present invention are all specific species, and the other species of lactobacillus plantarum, lactobacillus rhamnosus and bifidobacterium longum have no relieving effect on the swelling degree of the mouse ear, which also indicates that the species of special probiotics need to be screened for atopic dermatitis.

In addition, as compared with comparative examples 4 to 6, it can be seen that agaric polysaccharide and soyasaponin have a significant promoting effect on further relieving the swelling degree of ears of mice, and other polysaccharides, such as phellinus linteus polysaccharide, do not have the same relieving effect as agaric polysaccharide.

TABLE 1

2. Effect of immune inflammation index in atopic dermatitis mice

The results are shown in table 2, and it can be seen from table 2 that the embodiments of the technical scheme (example 1 to example 5) adopted by the invention all have a relatively significant effect of reducing the inflammatory factors (IL-4 and IL-5) related to the Th2 immune response, and the formula of the examples reduces the level of the inflammatory factors to be equivalent to the positive drug, even lower than the positive drug group, which indicates that the formula of the invention can be used for treating and/or relieving and/or assisting in treating atopic dermatitis.

The data of comparative examples 4 to 6 show that auricularia auricula polysaccharide can inhibit the release of inflammatory cytokines such as IL-4 and simultaneously inhibit the production of IgE antibodies, thereby reducing allergic reactions; soyasaponin helps to enhance the expression of IFN-gamma in skin tissue. In addition, the formula of the invention also obviously reduces the level of IgE and promotes the expression of IFN-gamma, thereby improving the capability of Th1 type immune response.

TABLE 2

Grouping	IL-4(pg/mL)	IL-5(pg/mL)	IgE(pg/mL)	IFN-γ(pg/mg)
					Example 1	2.8±0.1	3.8±0.2	17.1±0.1	98±1.6
Example 2	2.6±0.2	3.6±0.1	16.5±0.2	105±1.5
					Example 3	2.7±0.2	3.7±0.3	16.8±0.3	100±2.1
Example 4	2.8±0.3	3.8±0.2	16.9±0.1	82±1.6
					Example 5	3.0±0.2	4.0±0.2	17.0±0.2	85±1.5
Comparative example 1	5.2±0.1	5.5±0.1	18.5±0.2	88±2.3
					Comparative example 2	5.1±0.2	5.2±0.3	18.4±0.1	86±2.0
Comparative example 3	5.3±0.1	5.6±0.2	18.7±0.3	90±1.9
					Comparative example 4	5.6±0.3	5.8±0.2	19.0±0.2	89±1.8
Comparative example 5	5.3±0.2	5.5±0.3	18.6±0.2	95±2.2
					Comparative example 6	5.5±0.3	5.7±0.1	18.8±0.1	91±2.5
Blank group	4.3±0.2	4.4±0.2	17.8±0.2	110±1.5
					Model set	5.8±0.1	6.1±0.2	20.2±0.2	80±2.0
Positive drug group	2.7±0.2	3.8±0.3	17.5±0.3	94±1.8