CN116590201A - Bifidobacterium longum for inhibiting skin collagen fibrosis and application thereof - Google Patents
Bifidobacterium longum for inhibiting skin collagen fibrosis and application thereof Download PDFInfo
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- CN116590201A CN116590201A CN202310834609.8A CN202310834609A CN116590201A CN 116590201 A CN116590201 A CN 116590201A CN 202310834609 A CN202310834609 A CN 202310834609A CN 116590201 A CN116590201 A CN 116590201A
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Abstract
The invention provides a bifidobacterium longum for inhibiting skin collagen fibrosis and application thereof, wherein the bifidobacterium longum has a preservation number of CGMCC No.14168 and has the effect of inhibiting skin collagen fibrosis. The probiotic product prepared from the bifidobacterium longum can inhibit collagen fibrosis of skin and improve skin aging, and can be used as a probiotic for coordinating intestinal microbial balance, so that the effect of maintaining the health of a host organism is achieved, and the application prospect is wide.
Description
Technical Field
The invention belongs to the technical field of probiotics, and particularly relates to bifidobacterium longum for inhibiting skin collagen fibrosis and application thereof.
Background
Aging is caused by complex, multifactorial intrinsic and extrinsic factors that together manifest as reduced function in all organs of the body, with increased morbidity and mortality in the host. The skin acts as the first barrier to direct contact between the human body and the outside, accounting for about 10-15% of the body weight, and the average body surface area of an adult is about 1.6-2.0 square meters, which shows more rapid and more pronounced signs of aging over time than other organs. The over-emphasis of younger appearance importance in modern society has significantly increased people's anti-aging awareness. More importantly, the incidence of skin aging-related diseases is increasing, which also has a negative impact on quality of life. Therefore, effective measures capable of improving skin aging are urgently required.
Skin aging is characterized by structural changes in the form of reduced collagen content, clinically manifested as wrinkles, sagging, and a rough appearance due to loss of elasticity. Notably, collagen fibrosis is one of the important contributors to skin aging. The extracellular matrix (ECM) of the dermis layer of the skin is mainly composed of collagen fibers and elastic fibers, which are involved in a number of functions that coordinate skin homeostasis, including cell proliferation, hydration, hair growth and wound healing. Thus, prevention of collagen fibrosis, improvement of skin elasticity is critical for improving skin aging.
Bifidobacterium longum is one of the dominant bacteria in the host gut and plays an important role in body homeostasis. Recent studies have shown that bifidobacterium longum also appears to play a positive role in skin anti-ageing and cosmetic production etc. However, it is currently unclear how bifidobacterium longum exhibits these anti-aging effects on the skin. On the basis, the method is simple and convenient to operate, and is cheap and safe, and the skin aging can be improved by orally taking bifidobacterium longum, relieving collagen fibrosis of the skin and increasing skin elasticity.
Therefore, it is important to develop a probiotic that can exert an improving effect during skin aging, in particular against collagen loss during skin aging.
Disclosure of Invention
Aiming at the defects and actual demands of the existing related ways for improving skin aging, the invention provides bifidobacterium longum with the capability of improving skin collagen fibrosis and application thereof, the effect of bifidobacterium longum on improving skin aging is studied, skin aging indexes (moisture and elasticity), skin structure changes (epidermis thickness and dermis density), collagen content and collagen fibrosis conditions are detected through animal experiments, and the effect of the strain on improving skin aging is evaluated.
The invention aims at realizing the following technical scheme:
the invention provides a bifidobacterium longum BB68S separated from the intestinal tracts of healthy people, which is preserved in China general microbiological culture Collection center (CGMCC) of China Committee for culture Collection of microorganisms (China general microbiological culture Collection center) for 15 days in 2017, wherein the address of the preservation unit is North Chen West Lu No.1, 3 of the area of Chaoyang in Beijing, and the post code is 100101 of the institute of microorganisms of China academy of sciences, which is classified and named as bifidobacterium longumBifidobacterium longumThe preservation number is CGMCC NO. 14168.
The invention provides a method for culturing bifidobacterium longum, which is characterized in that bifidobacterium longum is inoculated in an MRS liquid culture medium and cultured for 18-24 hours at 30-37 ℃.
In another aspect, the invention provides the use of bifidobacterium longum in a product for improving skin ageing.
Preferably, the viable count of bifidobacterium longum in the product is not less than 1×10 6 CFU/mL。
Preferably, the product is any one of food, health care product, medicine or collagen-promoting agent.
In another aspect, the present invention provides a composition, collagen-promoting agent or related product comprising the above bifidobacterium longum.
The Bifidobacterium longum contained in the above product can improve collagen fibrosis in host skin.
Preferably, the viable count of Bifidobacterium longum is not less than 1×10 6 CFU/mL。
In another aspect, there is provided the use of a bifidobacterium longum of the invention to improve moisture and elasticity reduction in aged skin;
furthermore, the bifidobacterium longum can enable epidermis to be thinned and the symptom of loose dermis structure caused by aging to be relieved. Can be used for the application of thinning epidermis and improving the loose dermis structure.
On the other hand, the bifidobacterium longum provided by the invention can improve the Collagen fibrosis condition of aged skin, and is particularly characterized in that the expression quantity of type I Collagen (Collagen I) and type IV Collagen (Collagen IV) can be increased, the expression quantity of type III Collagen (Collagen III) and fibrosis marker protein alpha-SMA can be reduced, and finally the skin aging is improved.
The invention provides an application of bifidobacterium longum in improving aged skin, which is used for improving the levels of Collagen I and Collagen IV in the skin;
the invention provides an application of bifidobacterium longum for improving aged skin, which is used for reducing the generation of Collagen III and/or alpha-SMA.
The invention provides an application of bifidobacterium longum in improving aged skin, which can improve skin elasticity and hydration capacity by inhibiting skin fibrosis.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention successfully screens a strain of key enzyme (HAS 2) capable of up-regulating hyaluronic acid synthesis through flat plate coating and streak purification, thereby improving bifidobacterium longum generated by hyaluronic acid, and the preservation number is CGMCC NO. 14168.
(2) Bifidobacterium longum BB68S is capable of improving skin aging by alleviating collagen fibrosis in the skin of the host, thereby increasing skin elasticity.
(3) The probiotic product prepared by bifidobacterium longum BB68S can reduce the generation of Collagen III and alpha-SMA by improving the level of Collagen I and Collagen IV in skin, inhibit skin fibrosis, improve skin elasticity and hydration capacity, and finally improve skin aging.
Drawings
The skin physiological index levels of each group of fig. 1.
FIG. 2 each group of skin HE staining and Pinus massoniana trichromatic staining.
FIG. 3 skin sirius red staining and collagen fibrosis marker protein (. Alpha. -SMA) immunofluorescence staining of each group.
FIG. 4 western blotting and statistics of Collagen (Collagen I, collagen III and Collagen IV) in each skin group.
Detailed Description
The endpoints and any values of the ranges disclosed herein are not limited to the precise range or value, and are understood to encompass values approaching those ranges or values. For numerical ranges, one or more new numerical ranges may be found between the endpoints of each range, between the endpoint of each range and the individual point value, and between the individual point value, in combination with each other, and are to be considered as specifically disclosed herein.
The present invention will be described in detail by examples. It should be understood that the following examples are illustrative only of the present invention and are not intended to limit the present invention.
EXAMPLE 1 preparation of microbial culture
Preparation of bifidobacterium longum of the present invention: after the bifidobacterium longum is activated in the MRS culture medium for two generations, the bifidobacterium longum is inoculated in the MRS liquid culture medium for culturing 12 h in an inoculum size of 2 percent, the supernatant is removed by centrifugation (4000 rpm,10 min), after the thalli are washed twice by PBS, the thalli are resuspended in 0.9 percent physiological saline and the concentration of the bacterial suspension is regulated to be 1 multiplied by 10 when the thalli are irrigated 9 CFU/mL。
Example 2 animal model
SPF grade C57BL/6N (8 months old, male) mice were selected as experimental animals and purchased from Beijing Vetong Lihua experimental animal technology Co.
N-acetyl-N-2-formyl-5-methoxy kynurenine, N (1) -acetyl-N (2) -formyl-5-methoxykdynamic, AFMK, is a metabolite of melatonin, which was used as a positive control.
The animal experiments were divided into 4 groups, which were respectively a control group, a model group (natural aging group), an afmk+natural aging group, and a bifidobacterium longum+natural aging group, each group of 12 mice, and 48 mice in total.
Mouse feeding conditions: the temperature is 24 ℃, the illumination is controlled to be 12 h, the darkness is controlled to be 12 h, and the padding is replaced every three days. After one week of adaptive feeding, 48 mice were randomly divided into 4 groups of 12 mice each.
Control group (CON): 8 month old mice are directly obtained without any operation;
natural Aging group (Aging): placing 8-month-old mice under conventional conditions for normal feeding to 12 months of age;
AFMK intervention group (aging+afmk): mice were given AFMK 10 mg/kg/d by gavage every morning for a fixed period of time (7:00-8:00) starting at 8 months of age until 12 months of age.
Bifidobacterium longum intervention group (aging+b): mice were given 1X 10 by gavage every morning for a fixed time (7:00-8:00) starting at 8 months of age 9 CFU/mL bifidobacterium longum 0.2. 0.2 mL up to 12 months of age.
During the feeding period, four groups of mice were fed normal water, weighed once a week, and tested for skin physiology, including: skin moisture, elasticity, oil, pH, tension and gloss. After the intervention experiment is finished, the mice are fasted for 24 hours, blood of the mice is collected by taking eyeballs, and serum is collected after low-temperature centrifugation and frozen at-80 ℃ to be tested. The back skin of the mice is picked up by adopting aseptic operation and divided into two parts, one part is fixed in 4% paraformaldehyde, and the other part is frozen and stored in a refrigerator at-80 ℃ for detection. The intestinal tract was then dissected and the intestinal contents were removed for 16S rRNA detection of the intestinal flora.
Example 3 detection of skin Structure
The back skin of different groups of mice fixed in 4% paraformaldehyde is dehydrated by gradient alcohol, then embedded by paraffin, and finally cut into slices with the thickness of 5 mu m. The sections were used for HE staining, masson trichromatic staining, for observing skin epidermis thickness and dermis density.
The physiological index (moisture, elasticity, oil and pH) of the back skin of each group of mice is shown in FIG. 1, and the change in skin pH (C) and oil content (D) in the back skin of four groups of mice is not obvious. In addition, both skin moisture (a) and skin elasticity (B) were significantly reduced in the aged group as compared to the control group. However, after adding bifidobacterium longum and AFMK, the moisture and elasticity in the back skin are both significantly increased, but the improvement effect of bifidobacterium longum is more remarkable. The results show that bifidobacterium longum can improve the water content and the elasticity reduction in the aging skin during the aging process, thereby improving the skin aging.
As shown in FIG. 2, HE staining (A-D) and Masson trichromatic (E-H) staining demonstrated epidermal and dermal structures in the dorsal skin of four groups of mice. The statistics show that the epidermis thickness (I) and dermis density (J) of the skin in the aged group are significantly reduced compared to the control group. However, after adding bifidobacterium longum and AFMK, the epidermis thickness and dermis density increased significantly, but the improvement effect of bifidobacterium longum was more evident. The results indicate that the skin structure changes during skin aging, which is manifested by thinning of epidermis and loosening of dermis, while the addition of bifidobacterium longum can exert a good improving effect.
Example 4 detection of collagen fibrosis in skin
Skin sections from example 3 were used for sirius red staining and immunofluorescence staining to examine skin collagen fibrosis and expression of the intradermal collagen fibrosis marker protein α -SMA:
as shown in FIG. 3, sirius red (A-D) staining demonstrated collagen fibrosis in the skin of the back of four groups of mice, and immunofluorescence staining (E-H) demonstrated the expression of the fibrosis marker protein α -SMA in the skin. The combined statistics show that the skin fibrosis area (I) and the expression amount (J) of the alpha-SMA protein of the aging group are obviously increased compared with the control group. However, after adding bifidobacterium longum and AFMK, the expression level of fibrosis area and alpha-SMA protein is obviously reduced, but the improvement effect of bifidobacterium longum is more obvious. The results show that the expression level of alpha-SMA protein in dermis is gradually increased during skin aging to promote skin fibrosis, and the addition of bifidobacterium longum can effectively improve the process, and improve skin aging by inhibiting collagen fibrosis.
Example 5 detection of Collagen in skin (Collagen I, collagen III and Collagen IV)
The back skin of different groups of mice frozen at-80℃was examined for the expression of intradermal collagens (Collagen I, collagen III and Collagen IV) using western blotting.
As shown in FIG. 4, immunofluorescent staining showed the expression of three collagens (Collagen I, collagen III and Collagen IV) in the dermis layers of the back skin of four groups of mice. The combination statistics result shows that compared with the control group, the expression level of the Collagen I (A) and the Collagen IV (C) proteins in the skin of the aging group is obviously reduced, and the expression level of the Collagen III (B) proteins is obviously up-regulated. However, after adding bifidobacterium longum and AFMK, the expression level of the Collagen I and Collagen IV proteins is obviously increased, and the expression level of the Collagen III proteins is obviously reduced, but the improvement effect of bifidobacterium longum is more obvious. The results show that the contents of Collagen collgen I and collgen IV in dermis gradually decrease and the expression level of collgen III protein promoting Collagen fibrosis gradually increases in the skin aging process, and the addition of Bifidobacterium longum can effectively improve the process and improve skin aging by inhibiting Collagen fibrosis and Collagen degradation.
In conclusion, bifidobacterium longum can reduce the production of Collagen III and alpha-SMA by increasing the levels of Collagen I and Collagen IV in skin, inhibit skin fibrosis, improve skin elasticity and hydration capacity, and finally improve skin aging.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (10)
1. A bifidobacterium longum for inhibiting skin collagen fibrosis, characterized in that the bifidobacterium longum BB68S is classified and named as bifidobacterium longum Bifidobacterium longumThe microbial strain is preserved in China general microbiological culture Collection center (CGMCC) with the preservation number of CGMCC No.14168 and the preservation date of 2017, 05 and 15.
2. A method of culturing bifidobacterium longum as claimed in claim 1 wherein bifidobacterium longum is inoculated in MRS liquid medium and cultured at 30 to 37 ℃ for 18 to 24 hours.
3. Use of bifidobacterium longum as claimed in claim 1 in an oral product for improving skin ageing.
4. The use according to claim 3, wherein the viable count of bifidobacterium longum in the product is not less than 1 x 10 6 CFU/mL。
5. The use according to claim 3, wherein the product is any one of a food, a health product, a pharmaceutical product or a collagen-promoting regeneration agent.
6. A composition comprising the bifidobacterium longum of claim 1 or a broth fermentation thereof.
7. A microbial agent comprising the bifidobacterium longum of claim 1 or a microbial fluid fermentation thereof.
8. A product comprising bifidobacterium longum as claimed in claim 1 characterised in that the product is any one of a food, a health product, a pharmaceutical product or a collagen-promoting regeneration agent.
9. The composition of claim 6, the microbial inoculum of claim 7 and the use of the product of claim 8 for increasing the levels of Collagen i and Collagen iv in skin, reducing the production of Collagen iii and α -SMA, inhibiting skin fibrosis, and improving skin elasticity and hydration capacity.
10. The use according to claim 9, wherein the viable count of bifidobacterium longum is not less than 1 x 10 6 CFU/mL。
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| CN202310834609.8A CN116590201B (en) | 2023-07-10 | 2023-07-10 | Bifidobacterium longum for inhibiting skin collagen fibrosis and application thereof |
| NL2037741A NL2037741A (en) | 2023-07-10 | 2024-05-22 | Bifidobacterium longum capable of inhibiting skin collagen fibrosis and application thereof |
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|---|---|---|---|---|
| CN117551590A (en) * | 2024-01-10 | 2024-02-13 | 广州集妍化妆品科技有限公司 | Bifidobacterium longum subspecies infantis, microbial agent and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007073122A1 (en) * | 2005-12-23 | 2007-06-28 | Doosan Corporation | Composition for protecting and improving skin comprising the rice bran fermented by lactic acid bacteria |
| US20180311144A1 (en) * | 2015-10-15 | 2018-11-01 | Natura Cosméticos S.A. | Cosmetic composition having probiotic bacteria |
| CN113832061A (en) * | 2021-09-26 | 2021-12-24 | 广东一元兰欣生物科技有限公司 | Bifidobacterium longum NX-8 and application thereof in preparation of anti-aging drugs |
-
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2007073122A1 (en) * | 2005-12-23 | 2007-06-28 | Doosan Corporation | Composition for protecting and improving skin comprising the rice bran fermented by lactic acid bacteria |
| US20180311144A1 (en) * | 2015-10-15 | 2018-11-01 | Natura Cosméticos S.A. | Cosmetic composition having probiotic bacteria |
| CN113832061A (en) * | 2021-09-26 | 2021-12-24 | 广东一元兰欣生物科技有限公司 | Bifidobacterium longum NX-8 and application thereof in preparation of anti-aging drugs |
Non-Patent Citations (3)
| Title |
|---|
| 张晓妹等: ""长双歧杆菌BB68S体内及体外安全性评价"", 《中国奶牛》, vol. 2019, no. 11, pages 1 - 1 * |
| 李桂花等: ""长双歧杆菌BB68S对便秘人群的通便和调节肠道菌群作用"", 《中国奶牛》, vol. 2019, no. 12, pages 22 - 25 * |
| 闫昌誉等: ""二裂酵母(双歧杆菌)发酵产物在皮肤健康领域中的应用研究进展"", 《今日药学》, vol. 33, no. 8, pages 566 - 578 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117551590A (en) * | 2024-01-10 | 2024-02-13 | 广州集妍化妆品科技有限公司 | Bifidobacterium longum subspecies infantis, microbial agent and application thereof |
| CN117551590B (en) * | 2024-01-10 | 2024-04-05 | 广州集妍化妆品科技有限公司 | Bifidobacterium longum subspecies infantis, microbial agent and application thereof |
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| CN116590201B (en) | 2023-12-12 |
| NL2037741A (en) | 2024-06-11 |
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