Elephant-sourced bacillus licheniformis and application thereof

Abstract

The invention discloses an elephant source bacillus licheniformis and application thereof. The preservation number of the bacillus licheniformis strain L517 is GDMCC 67825, the bacillus licheniformis strain L517 is classified and named as bacillus licheniformis, and the preservation date is February 26, 2026. Research finds that the bacillus licheniformis has a remarkable treatment effect on acute colitis of mice; and the traditional Chinese medicine composition also has a remarkable treatment effect on mouse subcutaneous colon cancer. The anti-cancer mechanism is that the infiltration of GZMB < + > CD8 < + > cytotoxic T cells in a tumor microenvironment is remarkably promoted in vivo through a supernatant metabolite of a culture medium, and a tumor suppressor gene P53 can be directly up-regulated. It is found for the first time that the elephant intestinal tract source bacillus licheniformis with evolutionary disease-resistant advantages shows good anti-proctitis and colon cancer activity, and can become an effective medicine for treating proctitis and colon cancer in the future.

Description

Technical Field

[0001] This invention belongs to the field of biomedicine and microbiology, and specifically relates to the application of Bacillus licheniformis from elephants in the preparation of anti-inflammatory or anti-tumor drugs. Background Technology

[0002] Colorectal cancer (CRC) is one of the leading causes of death and morbidity among gastrointestinal malignancies worldwide, posing a serious public health problem. Despite the availability of various treatment options for CRC, such as surgery, chemotherapy, and targeted therapy, the prognosis for advanced CRC remains poor, and the disease is susceptible to treatment resistance and immune escape.

[0003] The pathogenesis of colorectal cancer (CRC) is a complex, multifactorial process, with chronic intestinal inflammation being a significant contributing factor. Increasing research elucidates a significant correlation between gut microbiota dysbiosis and its metabolites and inflammatory bowel disease (IBD) and CRC. Elephants possess unique cancer resistance, traditionally attributed to their multiple copies of the TP53 gene. However, as herbivores, elephants' intestines are colonized with a large number of cellulose-decomposing bacteria, such as Bacillus. The potential of these symbiotic bacteria in maintaining host immunity and fighting cancer remains largely unexplored. Therefore, developing novel, more effective anti-colorectal cancer microecological drugs from naturally resistant animals is a pressing technical challenge in this field. Summary of the Invention

[0004] The purpose of this invention is to provide a Bacillus licheniformis and its application in the preparation of anti-inflammatory or anti-tumor drugs.

[0005] To achieve the above objectives, the present invention adopts the following technical solution: a Bacillus licheniformis strain L517, with accession number GDMCC 67825, classified as Bacillus licheniformis, and accession date of February 26, 2026.

[0006] The application of the above-mentioned Bacillus licheniformis strain L517 in the preparation of antitumor drugs.

[0007] Furthermore, the tumor was colorectal cancer.

[0008] The application of the above-mentioned Bacillus licheniformis strain L517 in the preparation of anti-inflammatory drugs.

[0009] Furthermore, the anti-inflammatory drugs are anti-colitis drugs.

[0010] Furthermore, the Bacillus licheniformis strain L517 used in the drug is live Bacillus licheniformis from elephants, culture supernatant, or its metabolites.

[0011] Furthermore, the drug may also include pharmaceutically acceptable carriers or conventional edible excipients.

[0012] Furthermore, the drug is in oral dosage form.

[0013] This invention reveals that *Bacillus licheniformis* derived from elephants exhibits significant therapeutic effects on acute colitis in mice, restoring the Th17 / Treg cell balance; it also demonstrates significant therapeutic effects on colon cancer in mice, significantly increasing the infiltration of GZMB+CD8+ T cells within the tumor microenvironment. This invention is the first to discover that *Bacillus licheniformis* derived from elephants exhibits excellent anti-colitis and colon cancer activity, potentially becoming an effective drug for the future treatment of colitis and colon cancer. Attached Figure Description

[0014] Figure 1 This is an example of the acute DSS modeling process and the regulatory effect of Bacillus licheniformis treatment on the acute colitis phenotype and immune cells.

[0015] Figure 2 This is an example of the subcutaneous colorectal cancer modeling process and the effects of Bacillus licheniformis treatment on tumor growth inhibition and immune microenvironment regulation.

[0016] Figure 3 This is a diagram illustrating the effect of Bacillus licheniformis culture supernatant on inducing apoptosis in colorectal cancer cells and upregulating TP53 expression. Detailed Implementation

[0017] To enable those skilled in the art to better understand the present application, the technical solutions in the embodiments of the present application will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present application, and not all embodiments. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative effort should fall within the scope of protection of the present application.

[0018] It should be noted that the terms "comprising" and "having" and any variations thereof in the specification, claims and accompanying drawings of this application are intended to cover non-exclusive inclusion. For example, a process, method, system, product or device that includes a series of steps or units is not necessarily limited to those steps or units that are explicitly listed, but may include other steps or units that are not explicitly listed or that are inherent to such process, method, product or device.

[0019] Example 1: Treatment with Bacillus licheniformis can improve the DSS-induced acute colitis phenotype.

[0020] The process of establishing an acute colitis model was as follows: 8-week-old C57BL / 6 mice were randomly divided into a normal control group, a DSS group, and a treatment group. The treatment methods for each group were as follows:

[0021] Normal group: Drink pure water during the modeling process;

[0022] DSS group: Mice drank pure water containing 2.5% (w / v) DSS for 7 consecutive days, and then replaced with normal pure water;

[0023] Treatment group: The modeling protocol was the same as that of the DSS group. Throughout the experiment, mice were gavaged with 200 µL of bacterial suspension (2*10^9 CFU) every other day.

[0024] After modeling, mice were anesthetized, and their spleens and colons were harvested to measure colon length. Colon tissue was subjected to routine H&E staining. Flow cytometry was used to analyze single-cell suspensions of the spleen. Results showed that, compared to the DSS group, Bacillus licheniformis treatment significantly increased mouse survival rate and body weight, increased colon length, reduced splenomegaly, and improved intestinal pathology. Flow cytometry also showed that the treatment group significantly inhibited pro-inflammatory Th17 cells and increased the proportion of Foxp3+ regulatory T cells (Tregs) and anti-inflammatory M2 macrophages.

[0025] Example 2: Treatment with Bacillus licheniformis can inhibit the growth of subcutaneous colorectal cancer.

[0026] Subcutaneous colorectal cancer modeling process: 3-5×10 5 One MC-38 mouse colorectal cancer cell line was subcutaneously injected into the right ventral side of C57BL / 6 mice. The treatment methods for each group are as follows:

[0027] Control group: PBS was administered throughout the modeling process;

[0028] Treatment group: Throughout the modeling process, mice were orally administered Bacillus licheniformis (2*10^9 CFU) every other day.

[0029] After the model was established, mouse tumors were harvested and weighed. The tumor tissue was then prepared into a single-cell suspension using a MACS Octo dissociator for flow cytometry analysis.

[0030] The results showed that, compared with the control group, the tumor volume and weight of mice treated with Bacillus licheniformis were significantly reduced. Flow cytometry results revealed that Bacillus licheniformis significantly increased the percentage of GZMB+CD8+ cytotoxic T cells in the tumor area, thereby enhancing its ability to kill tumor cells.

[0031] Example 3: Bacillus licheniformis metabolites induce apoptosis and upregulate p53 expression in colorectal cancer cells.

[0032] Bacillus licheniformis was cultured in LB broth, the supernatant was collected and filtered for sterilization, and after resuspending, it was added to the culture medium of MC38 and HCT116 colorectal cancer cells at a volume ratio of 20% and incubated for 24 hours.

[0033] The results showed that the culture supernatant of Bacillus licheniformis significantly induced apoptosis in both cancer cell lines. LC-MS / MS metabolomics analysis revealed that the supernatant was enriched with metabolites such as valeric acid and nicotinamide mononucleotide (NMN). RT-qPCR analysis confirmed that the expression level of the tumor suppressor gene Trp53 in the treated MC38 cells was significantly higher than that in the untreated control group.

[0034] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. A Bacillus licheniformis strain L517, characterized in that: Its accession number is GDMCC 67825, its classification name is Bacillus licheniformis, and its accession date is February 26, 2026.

2. The use of Bacillus licheniformis strain L517 according to claim 1 in the preparation of antitumor drugs.

3. The application according to claim 2, characterized in that: The tumor is colorectal cancer.

4. The use of Bacillus licheniformis strain L517 according to claim 1 in the preparation of anti-inflammatory drugs.

5. The application according to claim 4, characterized in that: The anti-inflammatory drug is an anti-colitis drug.

6. The application according to any one of claims 2 to 5, characterized in that: The drug uses Bacillus licheniformis strain L517, which is live Bacillus licheniformis from elephants, culture supernatant, or its metabolites.

7. The application according to any one of claims 2 to 5, characterized in that: The drug may also include pharmaceutically acceptable carriers or conventional edible excipients.

8. The application according to any one of claims 2 to 5, characterized in that: The drug is in the form of an oral dosage form.

Images