CN114984059A

CN114984059A - Anti-colorectal cancer probiotic preparation and preparation method and application thereof

Info

Publication number: CN114984059A
Application number: CN202210482045.1A
Authority: CN
Inventors: 段相国; 苏春霞; 杨小娟; 马斌; 陈建; 田金花
Original assignee: Ningxia Medical University
Current assignee: Ningxia Medical University
Priority date: 2022-05-05
Filing date: 2022-05-05
Publication date: 2022-09-02

Abstract

The invention discloses a colorectal cancer resistant probiotic preparation and application thereof in preparation of medicaments for preventing and treating colorectal cancer, and belongs to the technical field of anti-tumor biomedicines. The anti-colorectal cancer probiotic preparation consists of the following components in parts by weight: 18-24% of lactobacillus plantarum powder, 18-24% of bifidobacterium adolescentis powder, 20-23% of stachyose, 10-16% of medlar extract, 2-5% of isomaltooligosaccharide and the balance of maltodextrin; wherein the fructus Lycii extract contains fructus Lycii polysaccharide as main ingredient. The anti-colorectal cancer probiotic preparation is prepared by mixing a medlar extract, probiotic plant lactobacillus capable of improving intestinal colony environment, bifidobacterium adolescentis and auxiliary materials of stachyose, isomaltooligosaccharide and maltodextrin, and can be used for remarkably inhibiting the tumor growth of CRC mice, the inhibition rate reaches 58%, and the mortality rate, especially the late-stage mortality rate of the CRC mice is remarkably reduced.

Description

Anti-colorectal cancer probiotic preparation and preparation method and application thereof

Technical Field

The invention belongs to the technical field of anti-tumor biological medicines, and particularly relates to a colorectal cancer resistant probiotic preparation as well as a preparation method and application thereof.

Background

Colorectal Cancer (CRC), one of the most common cancers threatening human life. Despite the great advances made in the prevention and treatment of CRC by surgery, chemoradiotherapy and biotherapy, the prevention and treatment of CRC still faces significant challenges.

The lycium barbarum polysaccharide is the main component of the traditional Chinese medicine lycium barbarum, and the content of the lycium barbarum polysaccharide in the lycium barbarum extract can reach more than 50%. Studies of Cao Yong show that lycium barbarum polysaccharide has obvious inhibition effect on SW480 cells, the inhibition effect has concentration dependence, and the inhibition rate is 20% -31%. And through electrophoresis experiments, the LBP can inhibit the expression of the anti-apoptosis gene Bcl-2 protein and induce apoptosis. Researches on Jiang Zheng Cao and the like show that the lycium barbarum polysaccharide has an inhibition effect on the growth of colorectal cancer tumor of mice, and may be related to inhibition of angiogenesis and up-regulation of BAI1 expression, but specific action mechanism is to be further clarified. However, when the lycium barbarum polysaccharides are used for treating colorectal cancer, on one hand, the unclear action mechanism is generally considered to be related to the enhancement of the immune function of the organism by the lycium barbarum polysaccharides. On the other hand, the lycium barbarum polysaccharides have single action and lower inhibition effect on the growth of colorectal cancer tumors due to various reasons, and the large dose of lycium barbarum polysaccharides is easy to aggravate the burden of the stomach, influence the digestion of intestines and stomach, and is unfavorable for the treatment of colorectal cancer.

Disclosure of Invention

Based on the above, the invention provides a probiotic preparation for resisting colorectal cancer, which aims to solve the technical problem that the suppression efficiency of small-dose lycium barbarum polysaccharide on colorectal cancer is low in the prior art.

The invention also provides application of the anti-colorectal cancer probiotic preparation in preparation of a medicine for preventing and treating colorectal cancer.

The technical scheme for solving the technical problems is as follows:

the anti-colorectal cancer probiotic preparation consists of the following components in parts by weight: 18-24% of lactobacillus plantarum powder, 18-24% of bifidobacterium adolescentis powder, 20-23% of stachyose, 10-16% of medlar extract, 2-5% of isomaltooligosaccharide and the balance of maltodextrin.

Preferably, the wolfberry polysaccharide content in the wolfberry extract is more than or equal to 35 percent by mass.

Preferably, the wolfberry polysaccharide content in the wolfberry extract is more than or equal to 50 percent by mass.

Preferably, the mass ratio of the lactobacillus plantarum powder to the bifidobacterium adolescentis powder is 1: 1.

The application of the anti-colorectal cancer probiotic preparation in preparing a medicament for preventing and treating colorectal cancer.

Preferably, the prepared medicament is used for inhibiting CD4 in spleen ⁺ CD25 ⁺ The proportion of Treg cells, promote the proportion of CD4+ IL-4+ Th2 cells and inhibit TIGIT molecules expressed by the cells, thereby promoting the proportion of CD19+ GL-7+ B cells and realizing the prevention and treatment of colorectal cancer.

Preferably, the type of the medicine is one of oral liquid, tablet, pill, granule, capsule, injection, drop pill, inhalant, syrup, gel or ointment.

Compared with the prior art, the invention has at least the following advantages:

the anti-colorectal cancer probiotic preparation is prepared by mixing a medlar extract (mainly comprising medlar polysaccharide), probiotics lactobacillus plantarum and bifidobacterium adolescentis capable of improving intestinal colony environment and auxiliary materials of stachyose, isomaltooligosaccharide and maltodextrin, and can be used for remarkably inhibiting the tumor growth of CRC mice, the inhibition rate reaches 58%, and the mortality rate, especially the late mortality rate of the CRC mice is remarkably reduced. Flow results show that the anti-colorectal cancer probiotic preparation passes the inhibition of CCD4 in spleen of RC mouse ⁺ CD25 ⁺ The proportion of Treg cells promotes CD4 ⁺ IL-4 ⁺ Th2 cell proportion and inhibiting its expression TIGIT molecule, thereby promoting CD19 ⁺ GL-7 ⁺ The proportion of B cells, thereby achieving the purpose of preventing and treating colorectal cancer.

Drawings

FIG. 1 is a product analysis test report sheet of an extract of Lycium barbarum used in an example of the present invention.

FIG. 2 is a graph comparing the sizes of the tumor volumes of the mice in each group.

Figure 3 is a line graph of mortality in mice as a function of time.

FIG. 4 is a graph showing the results of HE staining of mouse colorectal tissues.

FIG. 5 is a PCOA analysis chart of intestinal flora distribution in mice.

FIG. 6 is a Venn diagram of the intestinal flora distribution of mice.

Fig. 7 is a statistical result of the Venn diagram of the intestinal flora distribution shown in fig. 6.

Fig. 8 is the effect of the anti-colorectal cancer probiotic formulation on the immune microenvironment of CRC mice (a).

Fig. 9 is the effect of the anti-colorectal cancer probiotic formulation on the immune microenvironment of CRC mice (B).

FIG. 10 is a graph of the effect of anti-colorectal cancer probiotic formulation on the immune microenvironment of CRC mice (C)

Figure 11 is a graph of the effect of a probiotic formulation against colorectal cancer on apoptotic proteins in tumor tissue in CRC mice.

Detailed Description

It should be noted that the embodiments and features of the embodiments of the present invention may be combined with each other without conflict. The technical solutions of the present invention will be further described below with reference to the accompanying drawings of the embodiments of the present invention, and the present invention is not limited to the following specific embodiments.

It should be understood that the same or similar reference numerals in the drawings of the embodiments correspond to the same or similar parts. In the description of the present invention, it should be understood that if there are terms such as "upper", "lower", "front", "rear", "left", "right", "top", "bottom", etc., indicating orientations or positional relationships based on the orientations or positional relationships shown in the drawings, it is only for convenience of description and simplicity of description, but does not indicate or imply that the equipment or elements referred to must have a specific orientation, be constructed in a specific orientation, and be operated, and therefore, the terms describing the positional relationships in the drawings are only used for illustrative purposes and are not to be construed as limiting the patent, and the specific meanings of the terms will be understood by those skilled in the art according to specific situations.

In a specific embodiment, the anti-colorectal cancer probiotic preparation consists of the following components in parts by weight: 18-24% of lactobacillus plantarum powder, 18-24% of bifidobacterium adolescentis powder, 20-23% of stachyose, 10-16% of medlar extract, 2-5% of isomaltooligosaccharide and the balance of maltodextrin; wherein the fructus Lycii extract contains fructus Lycii polysaccharide as main ingredient.

For example, a probiotic formulation against colorectal cancer, consists of, by weight: 20% of lactobacillus plantarum powder, 20% of bifidobacterium adolescentis powder, 23% of stachyose, 16% of medlar extract, 5% of isomaltooligosaccharide and the balance of maltodextrin. Wherein the fructus Lycii extract contains fructus Lycii polysaccharide as main ingredient.

In the invention, the lactobacillus plantarum powder and the bifidobacterium adolescentis powder are prepared from a microbial inoculum obtained by a conventional method, for example, the bifidobacterium adolescentis powder can be selected from a microbial inoculum with the model number BAC30 of Jiangsu Weikang biotechnology limited, and the specification of the bifidobacterium adolescentis powder is 500 hundred million cfu/g. The lactobacillus plantarum powder can be selected from a bacterial agent of Jiangsu Weikang biological science and technology limited company with the model number of Lp90, and the specification of the lactobacillus plantarum powder is 3000 hundred million cfu/g.

The medlar extract serving as the main active component in the invention is a product obtained by medlar through methods of water extraction, alcohol precipitation and the like, and the medlar extract mainly contains medlar polysaccharide and possibly also contains carotenes, pigments, alkaloids and fatty acid substances. Preferably, the wolfberry polysaccharide content in the wolfberry extract is more than or equal to 35 percent by mass. In order to further ensure the treatment effect of the prepared anti-colorectal cancer probiotic preparation on colorectal cancer, the wolfberry polysaccharide content in the wolfberry extract is more than or equal to 50 percent by mass.

In the embodiment of the invention, the lycium barbarum polysaccharide powder from industry of lycium barbarum, Ningxia Volbifer, and the related technical index requirements and the actual detection results are shown in figure 1.

Preferably, the mass ratio of the lactobacillus plantarum powder to the bifidobacterium adolescentis powder is 1:1, so that the colony balance of the intestinal tract can be adjusted with the best effect.

In a further embodiment of the present invention, there is provided a use of the above-mentioned anti-colorectal cancer probiotic preparation for the preparation of a medicament for the prevention and treatment of colorectal cancer.

Preferably, the prepared medicament takes the anti-colorectal cancer probiotic preparation as a main active ingredient.

Further, the prepared medicine can inhibit CD4 in spleen ⁺ CD25 ⁺ Proportion of Treg cells, promoting CD4 ⁺ IL-4 ⁺ Th2 cell proportion and TIGIT molecule inhibiting its expression, thereby promoting CD19 ⁺ GL-7 ⁺ B cell ratio, and achieves prevention and treatment of colorectal cancer.

It is to be understood that the present invention is not limited to a specific type of probiotic preparation for colorectal cancer, and the type of the probiotic preparation is one of oral liquid, tablet, pill, granule, capsule, injection, drop pill, inhalant, syrup, gel or ointment, as the conditions allow.

The technical scheme and technical effects of the invention are further explained by specific experimental procedures.

In the present invention, the sources of the various reagents are as follows:

1. laboratory animal

SPF grade C57BL/6J male mice (age: 6 weeks; weight: 18-22g) were purchased from Beijing Huafukang Biotech GmbH (Beijing, China). All animal experiments were performed in strict compliance with the protocols of the Experimental animals center of Ningxia university of medicine, and were approved by the ethical Committee of Ningxia university of medicine (Yinchuan, China, approval No.: IACUC-NYLAC-2019-023).

2. Microbial inoculum and reagent

Both AOM and DSS were purchased from sigma aldrich (shanghai); AIN-93G feed is provided by broomcorn millet has tai (beijing) biotechnology limited; bifidobacterium adolescentis powder (model: BAC 30; specification: 500 hundred million cfu/g) and Lactobacillus plantarum powder (model: Lp 90; specification: 3000 million cfu/g) were purchased from Jiangsu Weikang Biotechnology Ltd; the fructus Lycii extract is obtained from fructus Lycii polysaccharide powder (more than 35%) of Bairuiyuan fructus Lycii Gmbycis; isomaltose hypgather, maltodextrin and stachyose were all purchased from Shanghai-derived leaf Biotech, Inc.

It is worth noting that the sources of the experimental animals, the microbial inoculum and the reagents are not limited in the invention on the premise of meeting the requirements of the experimental conditions of the invention.

Possibility and mechanism research of anti-colorectal cancer probiotic preparation for inhibiting colorectal cancer

1. Establishment of CRC model mouse

Grouping of animals: SPF grade C57BL/6J mice after two weeks of acclimation were randomly divided into 3 groups of 18 mice each. Respectively a normal mouse control group (NC group), a colorectal cancer mouse control group (RC group) and a colorectal cancer mouse probiotic preparation intervention group (CP group).

NC group only feeds normally;

after the RC group induces a mouse colorectal cancer model by an AOM/DSS method, the model is fed normally;

and (3) inducing a mouse colorectal cancer model by the CP group through an AOM/DSS method, normally feeding, and performing intragastric administration of an anti-colorectal cancer probiotic preparation. 6.6mg of the anti-colorectal cancer probiotic preparation was weighed, dissolved in 0.3ml of PBS and administered to the mice by gavage through a gavage needle. The stomach is drenched once every two days. The anti-colorectal cancer probiotic preparation is prepared by mixing the following components in parts by weight: 24% of lactobacillus plantarum powder, 24% of bifidobacterium adolescentis powder, 23% of stachyose, 16% of medlar extract, 5% of isomaltooligosaccharide and 8% of maltodextrin. The anti-colorectal cancer probiotic preparation is stored at a low temperature of-20 ℃ and is fully dissolved by normal saline when being used.

2. HE staining

Mice were sacrificed by cervical dislocation at week 13. After sacrifice, there are several major tasks: firstly, taking a spleen of a mouse, extracting mononuclear cells, and detecting the phenotype of related immune cells by adopting a flow cytometry method; next, the colorectal of the mouse was removed, washed clean, and the size of the tumor was measured with a vernier caliper. Then, taking down the tumor by using surgical scissors and tweezers, putting a part of the tumor into a freezing storage tube, and storing the marked part of the tumor in a liquid nitrogen tank for extracting tumor tissue protein; the other part was fixed in 4% paraformaldehyde for HE staining.

3. Westernblot detection of expression conditions of related proteins in colorectal cancer tissues of mice

Proteins were extracted from tumor tissues using a whole protein extraction kit (KeyGen Biotech co.ltd.) and then protein concentration was measured using a BCA protein quantification kit (KeyGen Biotech co.ltd.). 30ug of protein samples were electrophoresed by 10% SDS PAGE and then transferred to membranes by wet-transfer. After blocking with 5% skim milk powder, diluted GAPDH (1:2000cat. No. TA-08; BIOSS), BAX (1:1000cat. No.60267-1-Ig proteintech) and Bcl-2 antibody (1:1500cat. No.26593-1-APProteintech) were added and incubated overnight in a 4 degree refrigerator. The following day, the strips were incubated with goat anti-mouse or rabbit IgG horseradish peroxidase (HRP) conjugated secondary antibody (1:5,000; cat. nos. a21010 and a d a21020) for 1h at room temperature and detected by chemiluminescence, followed by analysis using ImageJ software.

4. Extraction of mononuclear cells

The spleen of the mouse was removed and excess tissue, fascia, was removed with scissors. After the spleen is ground, the ground suspension is filtered into a 15mL centrifuge tube by a 200-mesh filter screen, PBS is added to 3mL, the mixture is fully mixed and then added into the centrifuge tube containing a separation solution (Solarbio), and the mixture is centrifuged for 20 minutes at the rotating speed of 2000 r/min. Sucking the leucocyte layer, adding PBS with 5 times of volume, fully and uniformly mixing, centrifuging for 10 minutes at the rotating speed of 1500r/min, counting and dividing tubes.

5. Flow cytometry

The immune cells were detected by BD FACSCELESTITM flow cytometry, and the antibodies used in this experiment included surface antibodies AF488-conjugated anti-CD4 anti (BD Biosciences), FITC-conjugated anti-CD19 anti (BD Biosciences), and phytoerythrin-conjugated anti-TIGIT anti (BD Biosciences), as well as intrabodies. The intracellular antibodies include Phocoerythrorin-conjugated anti-forward box P3(anti-FOXP3) antibody (BD biosciences), BV421-conjugated anti-interleukin 4(anti-IL-4) antibody (BD biosciences), AF647-conjugated anti-GL-7antibody (BD biosciences). And incubating the surface antibody for 30 minutes in the dark, detecting on a machine, and dyeing after the intracellular antibody breaks the membrane.

6. Sequencing of 16S rDNA amplicon (16S rDNA amplification Sequencing)

The method of Sequencing mouse feces using 16S rDNA amplicon (16S rDNA amplification Sequencing) was based on double-ended Sequencing by Illumina NovaSeq Sequencing platform (Paired _ End), completed by Beijing NovoGene corporation (Chin, Beijing). Mainly comprises the following steps; extracting genome DNA, amplifying PCR, mixing PCR products, purifying, constructing a library, sequencing and analyzing sequencing data.

7. Discussion of Experimental results

Referring to fig. 2 and table 1, at the same time, the tumor volume of the CP mice intervened with the anti-colorectal cancer probiotic preparation is significantly smaller than that of the RC mice without any drug intervention, the maximum diameter of the tumor of the mice is also significantly reduced, and the inhibition rate calculated by the weight of the tumor of the mice is about 58.4% (inhibition rate ═ 1-CP group tumor weight/RC group tumor weight), which indicates that the anti-colorectal cancer probiotic preparation significantly inhibits the tumor growth of the CRC mice.

Referring to fig. 3 and table 2 together, as time goes on, the RC group mice and CP group mice died about 30 days, and their mortality rates were 16.7% and 5.6%, respectively, which means that the anti-colorectal cancer probiotic preparation can reduce the early-stage mortality rate of CRC mice. In about 60 days, the mice die intensively, and the comprehensive death rates of the RC group mice and the CP group mice respectively reach 66.7 percent and 44.4 percent. And over the period from 30 days to 60 days, the mortality rates of the RC group mice and the CP group mice were 60% and 41.2%, respectively, indicating that the anti-colorectal cancer probiotic formulation had limited effect on the mortality rate of the mid-term CRC mice. In about 90 days, the comprehensive death rate of the RC group mice reaches 77.8 percent, while the comprehensive death rate of the CP group mice is only 50 percent, which shows that the anti-colorectal cancer probiotic preparation obviously improves the survival rate of the CRC mice. Considering the mortality of mice over the period from 60 days to 90 days, the mortality of the RC group mice was 33.3% and the CP group was only 10%, which is probably closely related to the ability of the anti-colorectal cancer probiotic formulation to inhibit the growth of the mouse CRC mouse tumor.

(N:Number,ND:Numberofdeaths,NS:Numberofsurvivors)

With continued reference to fig. 4, the colorectal tissue of the mice in the RC group was severely damaged compared to the blank NC group, while the colorectal tissue of the mice in the CP group was significantly less damaged compared to the RC group, indicating that the anti-colorectal cancer probiotic preparation can effectively slow down the colorectal tissue damage of the CRC mice, and provide effective support for the body to resist the continuous development of colorectal cancer.

Referring to fig. 5-7, PCOA analysis shows that the intestinal flora of CRC mice intervened with the anti-colorectal cancer probiotic formulation is closer to that of normal mice as the intervention time is longer. The Venn plot indicates that at any time point, the number of OTUs shared by CRC mice and normal mice that were intervened with the anti-colorectal cancer probiotic formulation was always greater than the number of OTUs shared by non-intervened CRC mice and normal mice. The above results show that when CRC occurs, the intestinal flora of mice is disordered, and the anti-colorectal cancer probiotic preparation can effectively reverse the disorder.

Referring to fig. 8-10, flow results show that the anti-colorectal cancer probiotic preparation inhibited CD4 in spleen of CRC mice ⁺ CD25 ⁺ The proportion of Treg cells promotes CD4 ⁺ IL-4 ⁺ Th2 cell proportion and inhibiting its expression TIGIT molecule, thereby promoting CD19 ⁺ GL-7 ⁺ Proportion of B cells.

Please refer to fig. 11, which shows that the anti-colorectal cancer probiotic preparation promotes high expression of BAX protein in tumor tissues of CRC mice.

Specifically, based on the inhibition effect of the Treg cells in tumor immunity, the change of the Treg cells in CRC is firstly detected so as to determine whether the prevention and treatment effect of the anti-colorectal cancer probiotic preparation on CRC is related to the regulation of the proportion of the Treg cells. As shown in FIG. 8, the RC group mice CD4 compared with the normal mice ⁺ CD25 ⁺ The proportion of Treg cells was significantly increased, indicating CD4 ⁺ CD25 ⁺ Elevation of Treg cells is closely associated with the development of CRC. Thus, CD4 is inhibited ⁺ CD25 ⁺ The proportion of Treg cells may have a beneficial effect on the control of CRC. Analysis shows that the probiotic preparation can effectively inhibit the CD4 raised in CRC ⁺ CD25 ⁺ Treg cells, which show the intervention effect of the anti-colorectal cancer probiotic preparation and the down-regulation of CD4 ⁺ CD25 ⁺ The proportion of Treg cells is closely related.

As described above, it has been determined that the proportion of Treg cells is elevated in CRC and that intervention with a probiotic formulation against colorectal cancer is effective in suppressing the elevated Treg cells in CRC. It is well known that the suppressive effect of Treg cells in CRC is mainly accomplished by suppressing effector T cells. Therefore, next, it was investigated which types of effector T cells were mainly suppressed by Treg cells in CRC. As shown in FIG. 11, the RC group mice CD4 compared to the normal mice ⁺ IL-4+ Th2 cells were significantly reduced, indicating CD4 ⁺ IL-4 ⁺ The decrease in Th2 cells was closely associated with the development of CRC. Notably, the anti-colorectal cancer probiotic preparation can effectively promote CD4 ⁺ IL-4 ⁺ Th2 cell, demonstrating effects of probiotic intervention and promotion of CD4 ⁺ IL-4 ⁺ Th2 cells. In addition, CD4 was detected ⁺ IL-4 ⁺ A Th2 cell expressing TIGIT. TIGIT is an immunosuppressive molecule, similar to PD1, whose high expression inhibits effector T cell function. FIG. 11 shows that, after CRC occurs, CD4 ⁺ IL-4 ⁺ The expression of TIGIT of Th2 cells was also significantly increased, indicating that not only CD4 was involved in CRC ⁺ IL-4 ⁺ Th2 cells are decreased in number and their functions are also inhibited. The results show that the anti-colorectal cancer probiotic preparation can effectively up-regulate CD4 by down-regulating Treg cells ⁺ IL-4 ⁺ Th2 cells and downregulating TIGIT expression thereof.

Since the main function of Th2 cells is to promote the production of antibodies by B cells, humoral immunity is mediated. Therefore, the following studies have examined the change of B cells in CRC. CD19 ⁺ GL-7 ⁺ The B cell is a germinal center B cell, and can be mainly differentiated into plasma cells secreting antibodies, mediating humoral immunity, and also differentiated into memory B cells. As shown in FIG. 8, the RC group mice CD19 compared with the normal mice ⁺ GL-7 ⁺ Significant reduction of B cells, indicating CD19 ⁺ GL-7 ⁺ The reduction of B cells is closely related to the development of CRC. Notably, the probiotic formulation has the effect of promoting CD19 ⁺ GL-7 ⁺ The B cell capability shows that the anti-colorectal cancer probiotic preparation can play a beneficial role in CRC by down-regulating Treg and up-regulating Th2 cells and further up-regulating B cells.

Second, research on optimal material configuration of anti-colorectal cancer probiotic preparation

In order to further explore the optimal configuration of each material in the anti-colorectal cancer probiotic preparation, the following experimental processes are set:

after the SPF grade C57BL/6J mice adapted for two weeks are randomly divided into 10 groups of 18 mice each, and after the mouse colorectal cancer model is induced by the AOM/DSS method, the mice of each group are subjected to intragastric administration according to the material component distribution ratio in each example in the table 3, wherein the intragastric administration volume of each mouse is 0.3mL, the intragastric administration amount of each mouse is 6.6mg, and the intragastric administration is performed once every two days. The mortality rate of the mice in each group was counted 90 days after the administration, see table 3.

TABLE 3 Material configuration study of anti-colorectal cancer probiotic preparation

As can be seen from table 3, the probiotic preparation formed by the lactobacillus plantarum powder and/or the bifidobacterium adolescentis powder, stachyose, isomaltooligosaccharide and maltodextrin did not exhibit a strong inhibitory effect on the mortality of CRC mice although the intestinal flora distribution of CRC mice could be improved to some extent. Compared with the RC group, the composition formed by the medlar extract, stachyose, isomaltose hypgather and maltodextrin can obviously improve the survival rate of CRC mice, and the survival rate of the CRC mice in 60 days is improved by 15-25%.

In the first to sixth embodiments, the adopted lactobacillus plantarum powder, bifidobacterium adolescentis powder, medlar extract, stachyose, isomaltooligosaccharide and maltodextrin form the anti-colorectal cancer probiotic preparation, the mortality rate of the CRC mouse in 90 days is only 35% -50%, and is reduced by 25% -47.5% compared with the RC group, and is reduced by 10% -37% compared with the comparative ratio IV, which indicates that the synergistic effect of the lactobacillus plantarum powder, the bifidobacterium adolescentis powder and medlar extract further improves the inhibition efficiency of colorectal cancer, and improves the survival rate of the CRC mouse. And the inhibition efficiency of the colorectal cancer shows a certain dependence on the dosage of the medlar extract.

It should be understood that the above-described embodiments of the present invention are merely examples for clearly illustrating the present invention, and are not intended to limit the embodiments of the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. This need not be, nor should it be exhaustive of all embodiments. Any modification, equivalent replacement, and improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. The anti-colorectal cancer probiotic preparation is characterized by comprising the following components in parts by weight: 18-24% of lactobacillus plantarum powder, 18-24% of bifidobacterium adolescentis powder, 20-23% of stachyose, 10-16% of medlar extract, 2-5% of isomaltooligosaccharide and the balance of maltodextrin.

2. The anti-colorectal cancer probiotic preparation according to claim 1, wherein the content of lycium barbarum polysaccharides in the lycium barbarum extract is not less than 35% by mass.

3. The anti-colorectal cancer probiotic preparation according to claim 2, wherein the content of lycium barbarum polysaccharides in the lycium barbarum extract is not less than 50% by mass.

4. The anti-colorectal cancer probiotic formulation according to claim 1, wherein the mass ratio of the lactobacillus plantarum powder to the bifidobacterium adolescentis powder is 1: 1.

5. Use of a probiotic formulation against colorectal cancer according to any one of claims 1 to 4, for the preparation of a medicament for the prevention and treatment of colorectal cancer.

6. The use of claim 5, wherein the medicament is prepared by inhibiting CD4 in the spleen ⁺ CD25 ⁺ Proportion of Treg cells, promoting CD4 ⁺ IL-4 ⁺ Th2 cell proportion and TIGIT molecule inhibiting its expression, thereby promoting CD19 ⁺ GL-7 ⁺ B cell ratio, and achieves prevention and treatment of colorectal cancer.

7. The use of claim 5, wherein the drug is in the form of one of an oral liquid, tablet, pill, granule, capsule, injection, drop pill, inhalant, syrup, gel, or ointment.