CN112391321A

CN112391321A - Bifidobacterium bifidum NX-7 and application thereof in preparing medicament for treating ischemic diseases

Info

Publication number: CN112391321A
Application number: CN202011356472.2A
Authority: CN
Inventors: 张召; 郑康帝; 马燕; 赵英杰; 陈涛
Original assignee: Guangdong Longsee Medical Technology Co ltd; Foshan Langxin Biotechnology Co ltd
Current assignee: Foshan Langxin Biotechnology Co ltd; Guangdong Longsee Medical Technology Co ltd; Guangdong Yiyuan Lanxin Biotechnology Co ltd
Priority date: 2020-11-26
Filing date: 2020-11-26
Publication date: 2021-02-23
Anticipated expiration: 2040-11-26
Also published as: CN112391321B

Abstract

The invention discloses bifidobacterium bifidum NX-7 and application thereof in preparing a medicament for treating ischemic diseases, belonging to the technical field of microorganisms. The invention discloses bifidobacterium bifidum NX-7 with the preservation number of CGMCC No. 20115. The inactivated and non-inactivated fermented supernatant and bacterial suspension of bifidobacterium bifidum NX-7 can obviously promote the growth of the intestinal veins of zebra fish, obviously repair the damage of the vascular endothelial cell growth factor receptor inhibitor (PTK787) induced intestinal veins of zebra fish and obviously promote the damage repair of tail fins of the zebra fish, has the potential of promoting the regeneration and reconstruction of microcirculation, and shows the effect of being applied to the in vivo treatment of ischemic diseases. The bifidobacterium bifidum NX-7 disclosed by the invention has a huge potential application prospect in the aspect of treating ischemic diseases.

Description

Bifidobacterium bifidum NX-7 and application thereof in preparing medicament for treating ischemic diseases

Technical Field

The invention relates to the technical field of microorganisms, in particular to bifidobacterium bifidum NX-7 and application thereof in preparing a medicament for treating ischemic diseases.

Background

At present, the main clinical treatment means for treating ischemic cardiovascular diseases and cerebrovascular diseases are percutaneous coronary artery intervention, surgical bypass, drug thrombolysis and the like, and the treatment principle is mainly to rebuild blood perfusion and rescue ischemic tissues, however, the treatments have certain limitations. The cell proliferation, the angiogenesis and the repair are promoted by drug treatment or other intervention methods, so that the tissue regeneration and the microcirculation reconstruction are finally achieved, and a new idea is provided for the treatment of the cardiovascular and cerebrovascular diseases. Therefore, therapeutic angiogenesis has become one of the hot research spots for treating various ischemic diseases. Local administration of single angiogenesis promoting factors (VECF, bFGF) can effectively promote the angiogenesis of ischemic myocardium in animal experimental study, but the single angiogenesis promoting factors have short half-life in vivo and high price, and also have certain potential serious toxic and side effects, which undoubtedly bring many concerns and uneasiness to people. Therefore, the research direction for finding a treatment mode with reliable curative effect and small toxic and side effect becomes the main research direction.

Probiotics are defined as living microorganisms that improve the balance of the intestinal flora and may benefit various aspects of the physiological response of the host, including the immune system. In addition, probiotics have been extensively studied in the improvement of diabetes, obesity, and cardiovascular and cerebrovascular diseases. However, probiotics are currently less studied and used in ischemic diseases. 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, it is a problem to be solved by those skilled in the art to provide bifidobacterium bifidum NX-7 and its application in preparing a medicament for treating ischemic diseases.

Disclosure of Invention

In view of the above, the invention provides bifidobacterium bifidum NX-7 and application thereof in preparing a medicament for treating ischemic diseases.

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

the Bifidobacterium bifidum strain NX-7 has the preservation number of CGMCC No.20115, is preserved in the China general microbiological culture Collection center of China Committee for culture Collection of microorganisms (CGMCC for short), is collected at the institute of microbiology of China academy of sciences No.3, West Lu No. 1 Homeh, Ind. area of the Chaoyang region, has the preservation date of 2020, 06, 19 days, and is named as Bifidobacterium bifidum in classification.

Further, the bifidobacterium bifidum NX-7 is applied to the preparation of medicines for treating ischemic diseases.

Further, the bifidobacterium bifidum NX-7 is applied to preparing the medicine for promoting the growth of the intestinal veins of the zebra fish.

Further, the bifidobacterium bifidum NX-7 is applied to the preparation of the medicine for promoting the repair of the injury of the inferior intestinal vein of the zebra fish.

Further, the bifidobacterium bifidum NX-7 is applied to the preparation of the medicine for promoting the repair of the tail fin injury of the zebra fish.

Further, the bifidobacterium bifidum NX-7 is a non-inactivated bacterial suspension or a non-inactivated fermentation supernatant.

Further, the bifidobacterium bifidum NX-7 is inactivated bacterial suspension or inactivated fermentation supernatant.

The bifidobacterium bifidum NX-7 has the effects of promoting the growth of the intestinal veins of zebra fish, repairing PTK787 induced injury of the intestinal veins (SIVs) of zebra fish and promoting the repair of injury of the tail fin of zebra fish in vivo, has the potential of enhancing the self-repair capability of vascular tissues and shows good probiotic effect of treating ischemic diseases.

The bacterial strain NX-7 can obviously promote growth of the inferior intestinal vein of the zebra fish, obviously repair damage of PTK787 induced by the inferior intestinal vein of the zebra fish and also obviously promote repair of the tail fin damage of the zebra fish in vivo, and comprises inactivated bacterial strain NX-7, and fermentation supernatant (extracellular secretion) and bacterial suspension (thallus) which are not inactivated.

According to the technical scheme, compared with the prior art, the invention discloses and provides bifidobacterium bifidum NX-7 and application thereof in preparing a medicine for treating ischemic diseases, wherein the bifidobacterium bifidum NX-7 is obtained by separating and screening healthy infant feces, can remarkably promote growth of intestinal veins of zebra fish in vivo, remarkably repair PTK787 induced injury of the intestinal veins of the zebra fish and remarkably promote repair of injury of tail fins of the zebra fish, has the potential of promoting regeneration and reconstruction of microcirculation, shows the effect of being applied to in vivo treatment of ischemic diseases, and provides theoretical reference and guide basis for developing a probiotic preparation for treating the ischemic diseases by using the bifidobacterium bifidum NX-7.

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 bifidum NX-7 according to the invention;

FIG. 2 is a drawing showing the microscopic morphology observation of Bifidobacterium bifidum NX-7 after gram staining;

FIG. 3 is the attached drawing showing the effect of inactivated and non-inactivated fermented supernatant and bacterial suspension of Bifidobacterium bifidum NX-7 on the development of the inferior intestinal vein of zebra fish: imaging zebra fish intestinal vein budding;

FIG. 4 is the attached drawing showing the effect of inactivated and non-inactivated fermented supernatant and bacterial suspension of Bifidobacterium bifidum NX-7 on the development of the inferior intestinal vein of zebra fish: quantitatively analyzing the number of buds of the intestinal veins of the zebra fish;

FIG. 5 is the accompanying drawing showing the repairing effect of inactivated and non-inactivated fermented supernatant and bacterial suspension of Bifidobacterium bifidum NX-7 on PTK787 induced zebra fish inferior intestinal vein injury: a fluorescence imaging picture of the intestinal veins of the zebra fish;

FIG. 6 is the accompanying drawing of the repairing effect of inactivated and non-inactivated fermented supernatant and bacterial suspension of Bifidobacterium bifidum NX-7 on PTK787 induced zebra fish inferior intestinal vein injury: quantitatively analyzing the number of veins under the intestines of the zebra fish;

FIG. 7 is a graph showing the effect of inactivated and non-inactivated fermented supernatant and bacterial suspension of Bifidobacterium bifidum NX-7 on the repair of tail fin damage of zebra fish: an intuitive map of zebra fish tail fin damage repair;

FIG. 8 is a graph showing the effect of inactivated and non-inactivated Bifidobacterium bifidum NX-7 fermentation supernatant and bacterial suspension on the repair of tail fin damage of zebra fish: and (4) a statistical diagram of zebra fish tail fin damage repair.

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 bifidum NX-7

(1) Separation: after being diluted in a gradient way, the infant feces are respectively inoculated in a TPY solid culture medium, an MRS solid culture medium, a BHI solid culture medium and a BS solid culture medium, anaerobic culture is carried out for 48 hours at 37 ℃, and a single colony on a flat plate is selected and streaked 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.

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 fresh feces of healthy infants in Guangzhou city, Guangdong province is identified as bifidobacterium bifidum by morphological observation and 16S rDNA identification, and the 16S rDNA sequence of the strain is shown as SEQ ID NO. 3.

CTTACCATGCAGTCGAACGGGATCCATCGGGCTTTGCTTGGTGGTGAGAGTGGCGAACGGGTGAGTAATGCGTGACCGACCTGCCCCATGCTCCGGAATAGCTCCTGGAAACGGGTGGTAATGCCGGATGTTCCACATGATCGCATGTGATTGTGGGAAAGATTCTATCGGCGTGGGATGGGGTCGCGTCCTATCAGCTTGTTGGTGAGGTAACGGCTCACCAAGGCTTCGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACATTGGGACTGAGATACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGCAAGCCTGATGCAGCGACGCCGCGTGAGGGATGGAGGCCTTCGGGTTGTAAACCTCTTTTGTTTGGGAGCAAGCCTTCGGGTGAGTGTACCTTTCGAATAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTATCCGGATTTATTGGGCGTAAAGGGCTCGTAGGCGGCTCGTCGCGTCCGGTGTGAAAGTCCATCGCTTAACGGTGGATCTGCGCCGGGTACGGGCGGGCTGGAGTGCGGTAGGGGAGACTGGAATTCCCGGTGTAACGGTGGAATGTGTAGATATCGGGAAGAACACCGATGGCGAAGGCAGGTCTCTGGGCCGTCACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGGTGGACGCTGGATGTGGGGCACGTTCCACGTGTTCCGTGTCGGAGCTAACGCGTTAAGCGTCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGAAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGCGGATTAATTCGATGCAACGCGAAGAACCTTACCTGGGCTTGACATGTTCCCGACGACGCCAGAGATGGCGTTTCCCTTCGGGGCGGGTTCACAGGTGGTGCATGGTCGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTCGCCCCGTGTTGCCAGCACGTTATGGTGGGAACTCACGGGGGACCGCCGGGGTTAACTCGGAGGAAGGTGGGGATGACGTCAGATCATCATGCCCCTTACGTCCAGGGCTTCACGCATGCTACAATGGCCGGTACAGCGGGATGCGACATGGCGACATGGAGCGGATCCCTGAAAACCGGTCTCAGTTCGGATCGGAGCCTGCAACCCGGCTCCGTGAAGGCGGAGTCGCTAGTAATCGCGGATCAGCAACGCCGCGGTGAATGCGTTCCCGGGCCTTGTACACACCGCCCGTCAAGTCATGAAAGTGGGCAGCACCCGAAGCCGGTGGCCTAACCCCTTGTGGGATGGAGCCGTCTAAGTAGCTCA；SEQ ID NO.3。

The single colony of the strain is inoculated on a BS solid culture medium, and the strain grows well in an anaerobic way at 37 ℃, is spherical and white colony with regular edge (figure 1), and gram stain is positive (figure 2). The strain is preserved in China general microbiological culture Collection center (CGMCC), China institute of microbiology, national institute of sciences, China institute of sciences, No.3, West Lu 1, North Cheng, south China, Beijing, with a preservation date of 2020, 06, 19 days, is classified and named Bifidobacterium bifidum with a preservation number of CGMCC No. 20115.

EXAMPLE 2 preparation of Bifidobacterium bifidum NX-7 fermentation supernatant (extracellular secretion), bacterial suspension (thallus)

Activating and culturing Bifidobacterium bifidum NX-7, 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 10⁶Obtaining bacterial suspension (thallus) by CFU/mL; heating the fermentation supernatant (extracellular secretion) and bacterial suspension (thallus) at 121 deg.C for 15min to obtain heat-inactivated fermentation supernatant (extracellular secretion) and bacterial suspension (thallus).

Example 3 Effect of Bifidobacterium bifidum NX-7 on the development of the infraintestinal vein of Zebra fish

Development of the selectionPlacing healthy zebra fish Tg (fli1: EGFP) of 2hpf (hours post fertilization) in a 6-well cell culture plate, 15 strips/well, adding PBS in a blank group, adding inactivated or inactivated bacterial suspension in a bacterial suspension group, adding inactivated or inactivated fermented supernatant in a fermented supernatant group, changing the solution every 24h, after incubating for 72h at 28 ℃, anesthetizing the zebra fish by using tricaine, observing the growth condition of the intestinal veins and photographing under a fluorescence microscope, and then counting the number of the intestinal vein buds. Statistical processing of data and experimental data by using SPSS19.0 software

Data are presented using one-way analysis of variance. Each experimental group was compared to the blank group: p<0.05，**P<0.01，***P<0.005。

As can be seen from FIGS. 3 and 4, the blank group of zebrafish had almost no sprouting in the inferior intestinal vein (0.47. + -. 0.17 strips). The bacterial suspension of the bifidobacterium bifidum NX-7 which is not inactivated and the zebra fish of the fermentation supernatant group have the sprouting phenomenon in the intestinal veins (shown by an arrow in figure 3); meanwhile, the budding numbers of the intestinal veins of the zebra fish in the bifidobacterium bifidum NX-7 inactivated bacterial suspension and the fermented supernatant group are respectively 1.53 +/-0.19 strips and 1.80 +/-0.26 strips, and the difference of the average difference is obvious (P <0.01) compared with the blank group (0.47 +/-0.17 strips) (figure 4). In addition, the bifidobacterium bifidum NX-7 inactivated bacterial suspension and the zebra fish enterogastric vein of the fermentation supernatant group have the sprouting phenomenon (shown by an arrow in figure 3); meanwhile, the budding numbers of the intestinal veins of the zebra fish in the bifidobacterium bifidum NX-7 inactivated bacterial suspension and the fermented supernatant group are respectively 1.27 +/-0.15 and 1.60 +/-0.29, and the average difference is obvious (P <0.05) compared with the blank group (0.47 +/-0.17) (figure 4). Therefore, the results show that the suspension and the fermentation supernatant of the inactivated and non-inactivated bifidobacterium NX-7 can obviously promote the growth of new blood vessels in vivo, have the potential of promoting the regeneration and reconstruction of microcirculation and show the effect of treating ischemic diseases in vivo.

Example 4 Effect of Bifidobacterium bifidum NX-7 on PTK 787-induced repair of Damage to the infraintestinal vein of Zebra fish

Selecting healthy individuals that develop to 2hpf (hops fertilization)The zebrafish Tg (fli1: EGFP) is placed in a 24-well cell culture plate, 15 strips are placed in each well, PBS is added into a blank group, 0.03 mu g/mL of PTK787(Sigma-Aldrich) solution is added into a model group, a bacterial suspension group and a fermentation supernatant group, after incubation for 24 hours at 28 ℃, the PTK787 solution is abandoned, PBS is added into the blank group and the model group respectively, inactivated or inactivated bacterial suspension is added into the bacterial suspension group, inactivated or inactivated fermentation supernatant is added into the fermentation supernatant group, after incubation for 48 hours at 28 ℃, 1mL in each well, tricaine is used for anesthetizing the zebrafish, the growth of the intestinal veins is observed and photographed under a fluorescence microscope, and then the number of the intestinal veins is counted. Statistical processing of data and experimental data by using SPSS19.0 software

Data are presented using one-way analysis of variance. Each experimental group was compared to the model group: p<0.05，**P<0.01，***P<0.005。

As can be seen from FIGS. 5 and 6, the blank group of zebra fish has good development of the inferior intestinal vein and regular and ordered growth of blood vessels (10.73 +/-0.56 strips); the complete type inferior intestinal veins of the zebra fish in the model group (PTK787) are obviously reduced (shown by an arrow) (5.93 +/-0.36 strips), and the differences are obvious (P <0.005) compared with the blank group (10.73 +/-0.56 strips) (figure 6), which shows that the zebra fish inferior intestinal vein injury model is successfully constructed.

As can be seen from FIGS. 5 and 6, the bacterial suspension without inactivation of Bifidobacterium bifidum NX-7 and the zebra fish in the fermented supernatant group have good development of the inferior intestinal vein, regular and ordered growth of blood vessels, and budding phenomenon in the inferior intestinal vein (shown by an arrow in FIG. 5); meanwhile, the number of the intestinal veins of the zebra fish in the fermented supernatant group of the bifidobacterium bifidum NX-7 inactivated bacterial suspension is 8.00 +/-0.38 and 8.80 +/-0.49 respectively, and the average difference is obvious (P <0.01) compared with that of the model group (5.93 +/-0.36) (figure 6). In addition, the bifidobacterium bifidum NX-7 inactivated bacterial suspension and the zebra fish enterogastric veins of the fermented supernatant group are good in development, the blood vessels grow regularly and orderly, and the zebra fish enterogastric veins of the inactivated fermented supernatant group also have a budding phenomenon (shown by an arrow in fig. 5); meanwhile, the number of the intestinal veins of the zebra fish in the bifidobacterium bifidum NX-7 inactivated bacterial suspension and the fermented supernatant group is respectively 7.67 +/-0.40 and 8.00 +/-0.41, and the average difference is obvious (P <0.05) compared with that in the model group (5.93 +/-0.36) (figure 6). Therefore, the results show that the inactivated and non-inactivated bacterial suspension and the fermentation supernatant of the bifidobacterium bifidum NX-7 can obviously promote the damage repair of blood vessels in vivo, have the potential of promoting the regeneration and reconstruction of microcirculation and show the effect of treating ischemic diseases in vivo.

Example 5 Effect of Bifidobacterium bifidum NX-7 on the repair of injury to the tail fin of Zebra fish

Selecting normal-developing wild type AB line zebra fish (3dpf) and placing the zebra fish in a 6-hole cell culture plate, cutting the tail fin of the zebra fish by using a scalpel under a stereoscopic microscope, taking a picture at 0dpa (day post amplification) for recording, then transferring the zebra fish to a 96-hole cell culture plate, adding PBS (phosphate buffer solution) into a model group, adding non-inactivated or inactivated bacterial suspension into a bacterial suspension group, adding non-inactivated or inactivated fermented supernatant into a fermented supernatant group, wherein each hole is 200 mu L, each group is 20, after incubation is carried out to 3dpa, anesthetizing the zebra fish by using tricaine, and placing the zebra fish under the stereoscopic microscope for taking a picture for recording. Zebra fish tail fin lengths at 0dpa and 3dpa were counted as L1 and L2 using Image J software, respectively. The difference between L1 and L2 is the zebrafish tail fin regrowth length. Statistical processing of data and experimental data by using SPSS19.0 software

As can be seen from FIGS. 7 and 8, the tail fin of the zebra fish of the model group is not completely grown, and the regrowth length of the tail fin is 63.49 + -2.12 μm. The tail fins of the zebra fish of the bifidobacterium bifidum NX-7 inactivated bacterial suspension and the fermentation supernatant group are almost completely grown, the regrowth lengths of the tail fins are 83.06 +/-3.50 mu m and 91.74 +/-4.39 mu m respectively, and the difference is significant compared with that of a model group (63.49 +/-2.12 mu m) (P is less than 0.005). In addition, tail fins of zebra fish of the bifidobacterium bifidum NX-7 inactivated bacterial suspension and the fermentation supernatant group are almost completely grown, the regrowth lengths of the tail fins are 77.42 +/-2.89 mu m and 85.85 +/-3.64 mu m respectively, and the difference is significant compared with a model group (63.49 +/-2.12 mu m) (P < 0.05). Therefore, the results show that the suspension and the fermentation supernatant of the inactivated and non-inactivated bifidobacterium bifidum NX-7 can promote the damage repair of the tail fin of the zebra fish and have the potential of enhancing the self-repair capability of the vascular tissue.

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

<110> Guandong Nanxin medical science and technology Co., Ltd, Lanzhou, Chaishan, Inc

<120> Bifidobacterium bifidum NX-7 and application thereof in preparing medicine for treating ischemic diseases

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agagtttgat cctggctcag 20

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ggttaccttg ttacgactt 19

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cttaccatgc agtcgaacgg gatccatcgg gctttgcttg gtggtgagag tggcgaacgg 60

gtgagtaatg cgtgaccgac ctgccccatg ctccggaata gctcctggaa acgggtggta 120

atgccggatg ttccacatga tcgcatgtga ttgtgggaaa gattctatcg gcgtgggatg 180

gggtcgcgtc ctatcagctt gttggtgagg taacggctca ccaaggcttc gacgggtagc 240

cggcctgaga gggcgaccgg ccacattggg actgagatac ggcccagact cctacgggag 300

gcagcagtgg ggaatattgc acaatgggcg caagcctgat gcagcgacgc cgcgtgaggg 360

atggaggcct tcgggttgta aacctctttt gtttgggagc aagccttcgg gtgagtgtac 420

ctttcgaata agcgccggct aactacgtgc cagcagccgc ggtaatacgt agggcgcaag 480

cgttatccgg atttattggg cgtaaagggc tcgtaggcgg ctcgtcgcgt ccggtgtgaa 540

agtccatcgc ttaacggtgg atctgcgccg ggtacgggcg ggctggagtg cggtagggga 600

gactggaatt cccggtgtaa cggtggaatg tgtagatatc gggaagaaca ccgatggcga 660

aggcaggtct ctgggccgtc actgacgctg aggagcgaaa gcgtggggag cgaacaggat 720

tagataccct ggtagtccac gccgtaaacg gtggacgctg gatgtggggc acgttccacg 780

tgttccgtgt cggagctaac gcgttaagcg tcccgcctgg ggagtacggc cgcaaggcta 840

aaactcaaag aaattgacgg gggcccgcac aagcggcgga gcatgcggat taattcgatg 900

caacgcgaag aaccttacct gggcttgaca tgttcccgac gacgccagag atggcgtttc 960

ccttcggggc gggttcacag gtggtgcatg gtcgtcgtca gctcgtgtcg tgagatgttg 1020

ggttaagtcc cgcaacgagc gcaaccctcg ccccgtgttg ccagcacgtt atggtgggaa 1080

ctcacggggg accgccgggg ttaactcgga ggaaggtggg gatgacgtca gatcatcatg 1140

ccccttacgt ccagggcttc acgcatgcta caatggccgg tacagcggga tgcgacatgg 1200

cgacatggag cggatccctg aaaaccggtc tcagttcgga tcggagcctg caacccggct 1260

ccgtgaaggc ggagtcgcta gtaatcgcgg atcagcaacg ccgcggtgaa tgcgttcccg 1320

ggccttgtac acaccgcccg tcaagtcatg aaagtgggca gcacccgaag ccggtggcct 1380

aaccccttgt gggatggagc cgtctaagta gctca 1415

Claims

1. A Bifidobacterium bifidum NX-7 strain is characterized in that the preservation number is CGMCC No. 20115.

2. Use of bifidobacterium bifidum NX-7 as claimed in claim 1 in the manufacture of a medicament for the treatment of ischemic diseases.

3. Use of bifidobacterium bifidum NX-7 as claimed in claim 1 in the preparation of a medicament for promoting growth of the inferior intestinal vein of zebra fish.

4. Use of bifidobacterium bifidum NX-7 as claimed in claim 1 in the preparation of a medicament for promoting repair of the injury of the inferior intestinal vein of zebra fish.

5. Use of bifidobacterium bifidum NX-7 as claimed in claim 1 in the preparation of a medicament for promoting the repair of tail fin injury of zebra fish.

6. Bifidobacterium bifidum NX-7 as claimed in any of claims 1 to 5 is an inactivated bacterial suspension or an inactivated fermentation supernatant.

7. Bifidobacterium bifidum NX-7 as claimed in any of claims 1 to 5 is an inactivated bacterial suspension or an inactivated fermentation supernatant.