CN113832061A

CN113832061A - Bifidobacterium longum NX-8 and application thereof in preparation of anti-aging drugs

Info

Publication number: CN113832061A
Application number: CN202111129046.XA
Authority: CN
Inventors: 张召; 郑康帝; 陈涛
Original assignee: Foshan Langxin Biotechnology Co ltd; Guangdong Yiyuan Lanxin Biotechnology Co ltd
Current assignee: Foshan Langxin Biotechnology Co ltd; Guangdong Yiyuan Lanxin Biotechnology Co ltd
Priority date: 2021-09-26
Filing date: 2021-09-26
Publication date: 2021-12-24

Abstract

The invention discloses bifidobacterium longum NX-8 and application thereof in preparing an anti-aging medicament, belonging to the technical field of microorganisms. The invention discloses a bifidobacterium longum NX-8 with the preservation number of CGMCC No. 20116. The fermented supernatant and bacterial suspension of the bifidobacterium longum NX-8 can obviously reduce the level of active oxygen in the zebra fish body and obviously improve the activity of superoxide dismutase in the zebra fish body in an in-vivo oxidative stress model, and have the potential of resisting oxidation and delaying senescence when being applied to the body; and in a UVB ultraviolet ray induced human skin fibroblast photoaging model, the collagen I synthesis can be remarkably promoted, the matrix metalloproteinase-1 secretion can be remarkably inhibited, and the skin photoaging relieving effect is shown. The bifidobacterium longum NX-8 disclosed by the invention has huge potential application prospects in the aspects of resisting oxidation, delaying senescence and relieving skin photoaging.

Description

Bifidobacterium longum NX-8 and application thereof in preparation of anti-aging drugs

Technical Field

The invention relates to the technical field of microorganisms, in particular to bifidobacterium longum NX-8 and application thereof in preparing an anti-aging medicament.

Background

The aging process deteriorates the human body's function at multiple levels, resulting in a gradual decline in its ability to resist stress, injury and disease. In addition to changes in gene expression and metabolic control, senescence rates are also associated with the production of high levels of Reactive Oxygen Species (ROS). ROS are by-products of aerobic metabolism of cells, mainly comprising hydrogen peroxide (O)₂ ^2-) Singlet oxygen (a)¹O₂) Superoxide radical (O)₂ ^·-) Hydroxyl radical (. OH), etc., play an important role in the cell life cycle. Low concentrations of ROS can act as key signaling molecules within cells to regulate cell growth, proliferation and differentiation. However, the accumulation of ROS can severely damage cellular biological macromolecules such as proteins, lipids, DNA, etc., resulting in a variety of chronic diseases including atherosclerosis, arthritis, diabetes, neurodegenerative diseases, aging, skin aging, inflammatory bowel disease, etc. Increased levels of ROS have been shown to have a potentially critical role in the induction and maintenance of the cellular senescence process, as they not only disrupt cellular oxidation and inflammation homeostasis, but also accelerate telomere wear.

Ultraviolet radiation-induced ROS mediate a variety of cellular functions and alterations, including Matrix Metalloproteinase (MMP) secretion and extracellular matrix (ECM) degradation. Prolonged UVB uv exposure can lead to skin aging, resulting in wrinkle formation due to collagen and elastin fibrolysis and inhibition of procollagen synthesis. Collagen is located in the dermis layer of the skin in an amount of about 70%, and the major types are type I (85%), type III and type v, and fibrous collagen forms a network structure to provide skin tension and elasticity. MMPs mainly include collagenases (MMP-1 and-13) and gelatinases (MMP-2 and-9), which are capable of degrading all substances in the extracellular matrix, including collagen, elastin, glycoproteins, and the like. Thus, ROS-mediated oxidative stress-induced senescence can be said to be the most persistent but alterable factor that causes and drives the senescence program.

Senescent cells have been proposed as targets for intervention to delay senescence and its associated diseases or to improve disease therapy. Therapeutic intervention on aging cells may help to restore health and cure diseases that share underlying processes, rather than curing each disease individually and symptomatically. A large number of experimental studies show that antioxidants or free radical scavengers help to avoid the accumulation of ROS, have a protective effect on oxidative damage and can delay aging. However, most chemically synthesized or plant extracted antioxidant drugs are not recommended for long-term use due to potential adverse reactions. Therefore, the research direction for finding a treatment mode with reliable curative effect and small toxic and side effect becomes the main research direction. Compared with the traditional antioxidant drugs, probiotics with antioxidant effect are widely concerned due to the characteristics of small side effect, other probiotic effects and the like.

Probiotics are living microorganisms that, when administered in sufficient amounts, provide health benefits to the host. The use of probiotics has been associated with several health benefits including modulation of the intestinal flora, redox balance, immune response, cranial nerve modulation, etc. This explains the increasing use of probiotic-based dietary interventions for the treatment and prevention of different chronic diseases, in particular chronic diseases associated with stress and inflammation.

However, probiotics are currently less studied and used in antioxidants, delaying aging, and photoaging of skin. Meanwhile, the current international probiotic patent application focuses on the traditional research and development strong countries in the United states, the Japan and the Russia, and China lacks functional strains with independent intellectual property rights. Probiotic strains used by domestic production enterprises are imported for a long time, and foreign strains are not necessarily suitable for the gastrointestinal physiological conditions of residents in China. In addition, the function of the probiotics lacks strong scientific research evidence, and the popularization of the probiotics and the products thereof is seriously influenced. Based on the method, aiming at the deep excavation of the functions of the strain resources, the novel probiotic strain which has independent intellectual property rights, has specific functional properties and is suitable for the physiological characteristics of Chinese people is screened out, and the method is particularly important for improving the core competitiveness of probiotic production enterprises in China and promoting the development of probiotic products in China.

Therefore, the problem to be solved by the technical personnel in the field is to provide a bifidobacterium longum NX-8 and application thereof in preparing anti-aging medicaments.

Disclosure of Invention

In view of the above, the invention provides a bifidobacterium longum NX-8 and application thereof in preparing an anti-aging medicament.

In order to achieve the purpose, the invention adopts the following technical scheme:

a Bifidobacterium longum (Bifidobacterium longum) NX-8 with the preservation number of CGMCC No.20116 is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms (CGMCC for short), the institute of microbiology, China academy of sciences, No.3, West Lu No. 1, North Cheng, the area of south Korean, the preservation date is 2020, 06, 19 days, and is named as Bifidobacterium longum by classification.

Further, the bifidobacterium longum NX-8 is applied to the preparation of anti-aging medicaments.

Further, the bifidobacterium longum NX-8 is a bacterial suspension or a fermentation supernatant.

Further, the bacterial suspension or the fermentation supernatant is applied to reducing the ROS level in zebra fish bodies and improving the SOD activity in the bodies.

Further, the bifidobacterium longum NX-8 is applied to relieving skin photoaging.

Further, the bacterial suspension or the fermentation supernatant can obviously promote the synthesis of type I collagen (Col-I) and obviously inhibit the secretion of MMP-1 in a UVB ultraviolet ray induced human skin fibroblast photoaging model.

Further, the bifidobacterium longum NX-8 is applied to preparing antioxidant food, cosmetics and medicines.

The bifidobacterium longum NX-8 has the effects of reducing the ROS level in zebra fish bodies and improving the SOD activity in the zebra fish bodies in an in-vivo oxidative stress model, and shows good anti-oxidation and anti-aging probiotic effects; meanwhile, the strain can remarkably promote the synthesis of type I collagen (Col-I) and remarkably inhibit the secretion of MMP-1 in a UVB ultraviolet induced human skin fibroblast photoaging model, and shows a good effect of relieving skin photoaging.

The strain NX-8 which can obviously reduce the ROS level in zebra fish bodies and obviously improve the SOD activity in the zebra fish bodies in an in-vivo oxidative stress model, can also obviously promote the synthesis of I type collagen in a UVB ultraviolet induced human skin fibroblast photoaging model and obviously inhibit the secretion of MMP-1, and comprises a fermentation supernatant (extracellular secretion) and a bacterial suspension (thallus) of the strain NX-8.

According to the technical scheme, compared with the prior art, the invention discloses and provides a bifidobacterium longum NX-8 and application thereof in preparing an anti-aging medicament, wherein the bifidobacterium longum NX-8 is obtained by separating and screening excrements of elderly people with long lives in Ridge county of Musaceae, City, Guangdong province, has the potential of remarkably reducing the ROS level in zebra fish bodies, remarkably improving the SOD activity in the zebra fish bodies, remarkably promoting the synthesis of type I collagen and remarkably inhibiting the secretion of MMP-1 in a UVB ultraviolet human skin induced fibroblast photoaging model, and provides theoretical reference and guide basis for developing a probiotic preparation for resisting oxidation, delaying aging and relieving skin photoaging by utilizing the bifidobacterium longum NX-8.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 is a diagram showing the morphology of colonies formed by Bifidobacterium longum NX-8 according to the present invention;

FIG. 2 is a drawing showing the microscopic morphology observation of Bifidobacterium longum NX-8 after gram-staining;

FIG. 3 is a visual chart showing the effect of fermentation supernatant and bacterial suspension of Bifidobacterium longum NX-8 on the ROS level in a menadione-induced zebra fish oxidative stress model;

FIG. 4 is a statistical chart showing the effect of fermentation supernatant and bacterial suspension of Bifidobacterium longum NX-8 of the present invention on the ROS level in menadione-induced zebrafish oxidative stress model;

FIG. 5 is a drawing showing the effect of fermentation supernatant and bacterial suspension of Bifidobacterium longum NX-8 of the present invention on SOD activity in oxidative stress model of menadione-induced zebrafish;

FIG. 6 is a graph showing the effect of fermentation supernatant and bacterial suspension of Bifidobacterium longum NX-8 of the present invention on the synthesis of type I collagen (Col-I) in a model of ultraviolet UVB-induced photoaging of human skin fibroblasts;

FIG. 7 is a graph showing the effect of fermentation supernatant and bacterial suspension of Bifidobacterium longum NX-8 of the present invention on the secretion of MMP-1 in a model of ultraviolet UVB-induced photoaging of human skin fibroblasts.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

Example 1 isolation, identification and preservation of Bifidobacterium longum NX-8

(1) Separation: the method comprises the following steps of diluting the feces of the elderly with a gradient, respectively inoculating the feces into a PYG solid culture medium, a BHI solid culture medium, a BS solid culture medium and an MRS solid culture medium, carrying out anaerobic culture at 37 ℃ for 48h, and selecting a single colony on a plate to carry out streaking separation to obtain a pure colony. Inoculating pure bacterial colonies on the plate into a BS liquid culture medium, carrying out anaerobic culture at 37 ℃ for 12-16 h, adding 20% glycerol, and storing in a refrigerator at-80 ℃.

(2) And (3) strain morphological identification: the screened bacterial strains are observed under a microscope after gram staining, and gram positive bacteria are purple and gram negative bacteria are red.

(3) Molecular biological identification of the strains: extracting genome DNA of the obtained strain, amplifying a 16S rDNA full-length fragment by utilizing 16S rDNA universal primers 27F and 1492R through a PCR technology, and then sequencing to identify the strain species.

Wherein, the primer sequences of the universal primers 27F and 1492R are as follows:

27F：5’-AGAGTTTGATCCTGGCTCAG-3’；SEQ ID NO.1；

1492R：5’-GGTTACCTTGTTACGACTT-3’；SEQ ID NO.2。

the experimental results are as follows: the strain screened from feces of longevity elders in Mei Ling county of Guangdong province is identified as Bifidobacterium longum by morphological observation and 16S rDNA identification, wherein the strain NX-8 is identified as Bifidobacterium longum, and the 16S rDNA sequence is shown as SEQ ID NO. 3.

TCATCCCACAAGGGGTTAGGCCACCGGCTTCGGGTGCTGCCCACTTTCATGACTTGACGGGCGGTGTGTACAAGGCCCGGGAACGCATTCACCGCGACGTTGCTGATTCGCGATTACTAGCGACTCCGCCTTCACGCAGTCGAGTTGCAGACTGCGATCCGAACTGAGACCGGTTTTCAGGGATCCGCTCCGCGTCGCCGCGTCGCATCCCGTTGTACCGGCCATTGTAGCATGCGTGAAGCCCTGGACGTAAGGGGCATGATGATCTGACGTCATCCCCACCTTCCTCCGAGTTAACCCCGGCGGTCCCCCGTGAGTTCCCGGCATAATCCGCTGGCAACACGGGGCGAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACGACCATGCACCACCTGTGAACCCGCCCCGAAGGGAAGCCGTATCTCTACGACCGTCGGGAACATGTCAAGCCCAGGTAAGGTTCTTCGCGTTGCATCGAATTAATCCGCATGCTCCGCCGCTTGTGCGGGCCCCCGTCAATTTCTTTGAGTTTTAGCCTTGCGGCCGTACTCCCCAGGCGGGATGCTTAACGCGTTAGCTCCGACACGGAACCCGTGGAACGGGCCCCACATCCAGCATCCACCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTCGCTCCCCACGCTTTCGCTCCTCAGCGTCAGTAACGGCCCAGAGACCTGCCTTCGCCATTGGTGTTCTTCCCGATATCTACACATTCCACCGTTACACCGGGAATTCCAGTCTCCCCTACCGCACTCAAGCCCGCCCGTACCCGGCGCGGATCCACCGTTAAGCGATGGACTTTCACACCGGACGCGACGAACCGCCTAC；SEQ ID NO.3。

A single bacterial colony of the strain NX-8 is inoculated on a BS solid culture medium, the anaerobic growth is good at 37 ℃, the bacterial colony is white, round, smooth in surface, opaque and neat in edge (figure 1), and gram staining is positive (figure 2). The strain NX-8 is preserved in China general microbiological culture Collection center (CGMCC), is called CGMCC for short, and has the preservation date of 19 days in 2020 and 06 months in 19 days in Beijing, namely, the national institute of academy of sciences No.3, West Lu No. 1, North Cheng of the Korean district, and the preservation number of CGMCC No. 20116.

EXAMPLE 2 preparation of Bifidobacterium longum NX-8 fermentation supernatant (extracellular secretion), bacterial suspension (thallus)

Activating and culturing Bifidobacterium longum NX-8, inoculating into BS liquid culture medium, culturing at 37 deg.C for 15 hr, and adjusting the concentration of zymocyte to 1 × 10⁶Centrifuging at 4 deg.C and 6000r/min for 10min to obtain culture supernatant and thallus precipitate, and filtering the supernatant with 0.22 μm filter membrane to obtain fermentation supernatant (extracellular secretion); after the pellet was washed twice with PBS, the pellet was resuspended in PBS to adjust the cell concentration to 1X 10⁶CFU/mL gave a suspension (thallus). The fermentation supernatant (extracellular secretion) and bacterial suspension (bacterial cells) were used in examples 3 and 4.

Activating and culturing Bifidobacterium longum NX-8, inoculating into BS liquid culture medium, culturing at 37 deg.C for 15 hr, and adjusting the concentration of zymocyte to 1 × 10⁶Centrifuging for 10min at 4 ℃ and 6000r/min by CFU/mL to obtain culture supernatant and thallus precipitate, filtering the supernatant by a 0.22-micron filter membrane to obtain fermentation supernatant (extracellular secretion), and diluting the fermentation supernatant (extracellular secretion) to 50% (v/v) by using a DMEM culture medium (the DMEM culture medium is required for cell growth, and the DMEM culture medium is used for diluting the fermentation supernatant to 50% to provide a culture medium for HSF cell growth in the example 5); after the thalli sediment is washed twice by PBS, the thalli is re-cultured by DMEM mediumSuspending, adjusting cell concentration to 1 × 10⁶CFU/mL gave a suspension (thallus). The above 50% fermentation supernatant (extracellular secretion) and bacterial suspension (cells) were used in example 5.

Example 3 Effect of Bifidobacterium longum NX-8 on ROS levels in a Zebra Fish oxidative stress model

Reduced Glutathione (GSH), menadione, and dimethyl sulfoxide (DMSO) were obtained from Shanghai-derived leaf Biotech, Inc.; 2',7' -dichlorodihydrofluorescein diacetate (DCFH-DA) was purchased from Sigma-Aldrich.

Healthy wild-type AB line zebrafish that developed to 4dpf (days post fertilization) were picked and placed in 6-well cell culture plates, 20 fish per well. The experiment set up blank control group, model group, positive control group, sample (bacterial suspension, fermentation supernatant) intervention group, each group 20 fish. Adding PBS into a blank control group, adding PBS into a model group, adding GSH solution (100 mu M) into a positive control group, adding bacterial suspension into a bacterial suspension group, and adding fermentation supernatant into a fermentation supernatant group, wherein each hole is 2.5 mL; after incubation for 24h at 28 ℃, 2.5mL of PBS (1% DMSO) is added to the blank control group, and 6 μ M of menadione (menadione is prepared into 600 μ M stock solution by DMSO and then diluted into 6 μ M by PBS) is added to the model group, the positive control group, the bacterial suspension group and the fermentation supernatant group respectively, 2.5mL of each well; after incubation for 24h at 28 ℃, the solution is discarded, the zebra fish is washed for 3 times by PBS, 20 mu g/mL DCFH-DA solution is added, 3mL of the solution is added into each hole, after incubation for 1h at 28 ℃ in a dark place, the zebra fish is washed for 3 times by PBS, and the zebra fish is placed under a fluorescence microscope to observe the fluorescence intensity in vivo and take a picture for recording. Quantitative statistical analysis was performed on fluorescence intensity (S) in zebrafish using Image J software. ROS levels in zebrafish were calculated as follows:

statistical processing of data and experimental data by using SPSS 19.0 software

Data are presented using one-way analysis of variance. And blankComparison of control group:^###p<0.005; compared to the model set: p<0.01，***P<0.005。

The results are shown in FIGS. 3 and 4; as can be seen from FIGS. 3 and 4, the intensity of green fluorescence in zebra fish body reflects the level of ROS; compared with a blank control group, the intensity of green fluorescence in the zebra fish body of the model group is enhanced, which shows that the ROS level in the zebra fish body of the model group is increased; meanwhile, compared with a blank control group (100.00 +/-3.02%), the ROS level (189.65 +/-7.43%) in the zebra fish in the model group is obviously increased (p is less than 0.005), and the establishment of the zebra fish oxidative stress model is successful.

Compared with the model group, the green fluorescence intensity of the positive control Group (GSH) zebra fish in vivo is weakened, which indicates that the GSH can reduce the ROS level in the zebra fish in the menadione-induced zebra fish oxidative stress model; meanwhile, the ROS level in the zebra fish of the positive control group is 119.83 +/-6.85%, and the difference is obvious compared with that of a model group (189.65 +/-7.43%) (P is less than 0.005); therefore, GSH has an obvious antioxidant effect, consistent with clinical results. Compared with the model group, the green fluorescence intensity in vivo of the zebra fish in the fermentation supernatant and the bacterial suspension group of the bifidobacterium longum NX-8 is also weakened, which indicates that the ROS level in the zebra fish in the menadione-induced oxidative stress model of the zebra fish in the fermentation supernatant and the bacterial suspension of the bifidobacterium longum NX-8 can be reduced; the ROS levels in the fermentation supernatant and the bacterial suspension group zebra fish of the bifidobacterium longum NX-8 are 135.28 +/-7.51 percent and 163.63 +/-8.83 percent respectively, and the differences are significant (P <0.01) compared with the model group (189.65 +/-7.43 percent). Therefore, the results show that the fermentation supernatant and the bacterial suspension of the bifidobacterium longum NX-8 can both obviously reduce the ROS level in the zebra fish body in an in-vivo oxidative stress model, and show good effects of resisting oxidation and delaying aging.

Example 4 Effect of Bifidobacterium longum NX-8 on SOD Activity in Zebra Fish oxidative stress model

Healthy wild-type AB line zebrafish that developed to 4dpf (days post fertilization) were picked and placed in 6-well cell culture plates, 20 fish per well. The experiment set up blank control group, model group, positive control group, sample (bacterial suspension, fermentation supernatant) intervention group, each group 20 fish. Adding PBS into a blank control group, adding PBS into a model group, adding GSH solution (100 mu M) into a positive control group, adding bacterial suspension into a bacterial suspension group, and adding fermentation supernatant into a fermentation supernatant group, wherein each hole is 2.5 mL; after incubation for 24h at 28 ℃, 2.5mL of PBS (1% DMSO) is added to the blank control group, and 6 μ M of menadione (menadione is prepared into 600 μ M stock solution by DMSO and then diluted into 6 μ M by PBS) is added to the model group, the positive control group, the bacterial suspension group and the fermentation supernatant group respectively, 2.5mL of each well; after incubation for 24h at 28 ℃, discarding the solution, washing the zebra fish for 3 times by using PBS, collecting the zebra fish to a 1.5mL centrifuge tube, wherein each tube contains 50mg of zebra fish, and each experimental group contains 6 tubes; after the water in the centrifuge tube was blotted dry, 250. mu.L of buffer solution (buffer solution of superoxide dismutase (SOD) detection kit) was added. Treating the centrifuge tube with ultrasonicator in ice bath at an interval of 8s for 5s, ultrasonicating for 10 times, centrifuging at 12000 Xg for 10min at 4 deg.C, and collecting supernatant. SOD activity of each group was detected using a superoxide dismutase (SOD) detection kit (Sigma-Aldrich Co.).

Data are presented using one-way analysis of variance. Compared to the blank control group:^###p<0.005; compared to the model set: p<0.01，***P<0.005。

The results are shown in FIG. 5; as can be seen from FIG. 5, compared with the blank control group (3.80. + -. 0.55U/mg), the SOD activity (0.84. + -. 0.09U/mg) in the zebra fish body of the model group is significantly reduced (p is less than 0.005), which indicates that the establishment of the zebra fish oxidative stress model is successful.

The SOD activity in the zebra fish body of the positive control group is 3.01 plus or minus 0.27U/mg, and the difference is obvious (P is less than 0.005) compared with the model group (0.84 plus or minus 0.09U/mg), which indicates that the GSH has obvious antioxidation and is consistent with the clinical result. The SOD activities in the fermentation supernatant and the bacterial suspension of the bifidobacterium longum NX-8 in zebra fish are respectively 2.56 +/-0.27U/mg and 2.31 +/-0.21U/mg, and the differences are obvious (P is less than 0.01) compared with the model group (0.84 +/-0.09U/mg). Therefore, the results show that the fermentation supernatant and the bacterial suspension of the bifidobacterium longum NX-8 can obviously improve the SOD activity in zebra fish bodies, enhance the capability of the bodies to remove free radicals and show good effects of resisting oxidation and delaying senescence in-vivo oxidative stress models.

Example 5 Effect of Bifidobacterium longum NX-8 on type I collagen and MMP-1 content in HSF photoaging model of human skin fibroblasts

Subjecting HSF cells in logarithmic growth phase to pancreatin digestion and blow beating to obtain cell suspension, and regulating cell concentration to 5 × 10⁴One cell/mL, and the number of cells was about 1X 10 in 24-well plates⁴One/hole, put at 37 ℃ and 5% CO₂After incubation in the incubator for 24h, the cells were washed 3 times with sterile PBS; adding 1mL of sterile PBS into each hole, wherein a blank control group does not need UVB ultraviolet irradiation, and a model group and a sample (bacterial suspension and fermentation supernatant) pre-drying group are placed in a CL-1000 ultraviolet crosslinking instrument for irradiation (144mJ/cm) at 302nm for 40 seconds and then immediately washed by the sterile PBS for 3 times; then adding a DMEM culture medium into the blank control group, adding a DMEM culture medium into the model group, adding a bacterial suspension into the bacterial suspension group, adding 50% of fermentation supernatant into the fermentation supernatant group, continuously incubating for 48 hours, collecting cell supernatant, and detecting the secretion of type I collagen (Col-I) and MMP-1 by using an ELISA kit (Nanjing herbaceous biotechnology limited). Each group is respectively provided with 6 multiple holes. Statistical processing of data and experimental data by using SPSS 19.0 software

Data are presented using one-way analysis of variance. Compared to the blank control group:^###p<0.005; compared to the model set: p<0.05，**P<0.01。

The results are shown in FIGS. 6 and 7; as can be seen from FIGS. 6 and 7, compared with the blank control group (Col-I: 26.70 + -0.46 ng/mL, MMP-1: 4.20 + -0.16 ng/mL), the model group has a significant decrease in Col-I (15.46 + -1.05 ng/mL) and a significant increase in MMP-1(11.61 + -1.07 ng/mL) (p <0.005), indicating that the UVB ultraviolet ray induced human skin fibroblast photoaging model is successfully established.

The concentrations of the Col-I of the fermentation supernatant and the bacterial suspension group of the bifidobacterium longum NX-8 are respectively 20.99 +/-0.62 ng/mL and 19.02 +/-0.97 ng/mL, and the average difference is obvious (P is less than 0.05) compared with that of a model group (15.46 +/-1.05 ng/mL); in addition, the concentrations of MMP-1 in the fermentation supernatant and the bacterial suspension group of Bifidobacterium longum NX-8 are respectively 6.41 +/-0.86 ng/mL and 8.23 +/-0.55 ng/mL, and the average difference is significant (P <0.05) compared with the model group (11.61 +/-1.07 ng/mL). Therefore, the results show that the fermentation supernatant and the bacterial suspension of the bifidobacterium longum NX-8 can obviously promote the synthesis of type I collagen and obviously inhibit the potential of MMP-1 secretion in a UVB ultraviolet induced human skin fibroblast photoaging model, and show good effect of relieving skin photoaging.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Sequence listing

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<120> bifidobacterium longum NX-8 and application thereof in preparation of anti-aging drugs

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cgactccgcc ttcacgcagt cgagttgcag actgcgatcc gaactgagac cggttttcag 180

ggatccgctc cgcgtcgccg cgtcgcatcc cgttgtaccg gccattgtag catgcgtgaa 240

gccctggacg taaggggcat gatgatctga cgtcatcccc accttcctcc gagttaaccc 300

cggcggtccc ccgtgagttc ccggcataat ccgctggcaa cacggggcga gggttgcgct 360

cgttgcggga cttaacccaa catctcacga cacgagctga cgacgaccat gcaccacctg 420

tgaacccgcc ccgaagggaa gccgtatctc tacgaccgtc gggaacatgt caagcccagg 480

taaggttctt cgcgttgcat cgaattaatc cgcatgctcc gccgcttgtg cgggcccccg 540

tcaatttctt tgagttttag ccttgcggcc gtactcccca ggcgggatgc ttaacgcgtt 600

agctccgaca cggaacccgt ggaacgggcc ccacatccag catccaccgt ttacggcgtg 660

gactaccagg gtatctaatc ctgttcgctc cccacgcttt cgctcctcag cgtcagtaac 720

ggcccagaga cctgccttcg ccattggtgt tcttcccgat atctacacat tccaccgtta 780

caccgggaat tccagtctcc cctaccgcac tcaagcccgc ccgtacccgg cgcggatcca 840

ccgttaagcg atggactttc acaccggacg cgacgaaccg cctac 885

Claims

1. Bifidobacterium longum (Bifidobacterium longum) NX-8 is characterized in that the preservation number is CGMCC No. 20116.

2. Use of bifidobacterium longum NX-8 as claimed in claim 1 in the manufacture of an anti-ageing medicament.

3. The use of bifidobacterium longum NX-8 in the preparation of an anti-aging medicament as claimed in claim 2, wherein the bifidobacterium longum NX-8 is a bacterial suspension or a fermentation supernatant.

4. Use of bifidobacterium longum NX-8 as claimed in claim 1 for the relief of photoaging of the skin.

5. The use of Bifidobacterium longum NX-8 in the alleviation of skin photoaging as claimed in claim 4, wherein the Bifidobacterium longum NX-8 is a bacterial suspension or a fermentation supernatant.

6. Use of bifidobacterium longum NX-8 as claimed in claim 1 for the preparation of antioxidant foods, cosmetics, pharmaceuticals.

7. The use of Bifidobacterium longum NX-8 in the preparation of antioxidant foods, cosmetics, pharmaceuticals as claimed in claim 6, wherein the Bifidobacterium longum NX-8 is a bacterial suspension or a fermentation supernatant.