Bifidobacterium animalis lactis and use thereof in the preparation of a medicament for the treatment of graves' disease

Abstract

The present application relates to the field of biological medical technology, and in particular to animal bifidobacterium lactis and its application in the preparation of a drug for treating Graves' disease. The animal bifidobacterium lactis is named as animal bifidobacterium lactis (Bifidobacterium animalis lactis) NCU-42, which has been preserved in the China General Microbiological Culture Collection Center, located at No. 1, Beichen West Road, Hua-yuan District, Beijing, on April 17, 2025, and the preservation number is CGMCC No. 34254. The animal bifidobacterium lactis NCU-42 has probiotic properties and can improve hyperthyroidism and Th9 / Treg cell immune imbalance, thereby preventing or treating Graves' disease. Bifidobacterium animalis subsp. lactis ​ ​

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to Bifidobacterium lactis subsp. animalis and its application in the preparation of drugs for treating Graves' disease. Background Technology

[0002] Graves' disease (GD), a core type of autoimmune thyroid disease, is essentially an organ-specific immune disorder mediated by thyrotropin receptor antibody (TRAb). Typical features include thyrotoxicosis, diffuse goiter, and in some patients, thyroid-associated ophthalmopathy. Rarely, it is accompanied by a characteristic skin lesion—pretibial myxedema. Current clinical interventions for GD include three basic therapies: antithyroid drug therapy, radioactive iodine therapy, and thyroidectomy.

[0003] Antithyroid drugs are mainly thionamide compounds. Among them, methimazole (MMI) and propylthiouracil (PTU) block hormone synthesis by inhibiting thyroid peroxidase activity and are suitable for patients with mild to moderate goiter, mild hyperthyroidism symptoms, low TRAb levels, severe eye disease, or pregnancy. Although antithyroid drugs are the first choice, there are concerns about the potential for serious complications such as acute liver failure in the early stages of treatment, a relapse rate of up to 50% after remission in half of the patients, and the unclear immune regulation mechanism and molecular basis of remission.

[0004] Radioactive iodine therapy destroys thyroid tissue using beta rays, achieving a cure rate of up to 80%, but requires lifelong thyroid hormone replacement therapy. Regarding safety assessments, the latest meta-analysis shows no significant increase in overall cancer risk, but the potential upward trend in the incidence of thyroid, gastric, and kidney cancer warrants attention, suggesting a need for more comprehensive evidence-based medicine to support clinical decisions.

[0005] Surgical intervention, through thyroidectomy, achieves a radical cure with a success rate of nearly 100%. It is suitable for patients with significant thyroid hyperplasia, compressive symptoms, or suspected malignancy. However, the inevitability of permanent hypothyroidism after surgery limits its application. Summary of the Invention

[0006] In view of this, the present invention provides Bifidobacterium lactis subsp. animalis and its application in the preparation of drugs for treating Graves' disease, thereby solving at least one problem existing in the prior art.

[0007] In a first aspect, the present invention provides a strain of Bifidobacterium lactis (Bifidobacterium animalis subsp. lactis). Bifidobacterium animalis subsp. lactisThe above-mentioned Bifidobacterium lactis subspecies NCU-42 has been deposited at the China General Microbiological Culture Collection Center (CGMCC) at No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing. The deposit date is April 17, 2025, and the accession number is CGMCC No. 34254.

[0008] The 16S rRNA sequencing results of the above-mentioned Bifidobacterium lactis subspecies NCU-42 are shown in SEQ ID No. 1, which is as follows:

[0009]

[0010] The aforementioned *Bifidobacterium lactis* subsp. NCU-42 exhibits probiotic properties, demonstrating strong acid and bile salt resistance, inhibition of pathogenic bacteria, and antioxidant capacity, as well as non-hemolytic safety. Furthermore, *Bifidobacterium lactis* subsp. NCU-42 can improve thyroid function and histopathological progression in mice, and can be used to prepare drugs for treating Graves' disease.

[0011] Secondly, the present invention provides the use of the above-mentioned animal Bifidobacterium lactis subsp. NCU-42 in the preparation of a drug for the prevention or treatment of Graves' disease.

[0012] Thirdly, the present invention provides a drug for the prevention or treatment of Graves' disease, comprising the aforementioned Bifidobacterium lactis subsp. NCU-42.

[0013] In some optional embodiments, the dosage form of the above-mentioned drug for preventing or treating Graves' disease is a liquid formulation.

[0014] In some alternative embodiments, the aforementioned medicaments for the prevention or treatment of Graves' disease may also include pharmaceutically acceptable excipients.

[0015] In some alternative embodiments, the pharmaceutically acceptable excipients described above are water or physiological saline.

[0016] In some optional embodiments, the concentration of the above-mentioned Bifidobacterium animalis subsp. lactis NCU-42 is 0.1 × 10⁻⁶. 9 CFU / mL - 10 × 10 9 CFU / mL. Preferably, the concentration of the above-mentioned Bifidobacterium animalis subsp. lactis NCU-42 is 1×10⁻⁶ CFU / mL. 9 CFU / mL - 5 × 10 9 CFU / mL. Most preferably, the concentration of the above-mentioned Bifidobacterium animalis subsp. lactis NCU-42 is 1×10⁻⁶ CFU / mL. 9 CFU / mL.

[0017] Due to the adoption of the above technical solutions, the embodiments of the present invention have the following beneficial effects: a strain of Bifidobacterium lactis NCU-42 is provided, which has probiotic properties and can improve hyperthyroidism and Th9 / Treg cell immune imbalance, thereby preventing or treating Graves' disease. Attached Figure Description

[0018] Figure 1 This is a phylogenetic tree diagram of Bifidobacterium lactis subsp. NCU-42 in an embodiment of the present invention.

[0019] Figure 2This is a growth curve diagram of Bifidobacterium lactis subsp. NCU-42 in an embodiment of the present invention.

[0020] Figure 3 The results of the acid resistance test of Bifidobacterium lactis subsp. NCU-42 in the embodiments of the present invention are shown.

[0021] Figure 4 The results of the bile salt tolerance test of Bifidobacterium lactis subsp. NCU-42 in the embodiments of the present invention are shown.

[0022] Figure 5 The results of the antioxidant and reducing abilities of Bifidobacterium lactis subsp. NCU-42 in the embodiments of the present invention are shown.

[0023] Figure 6 The results of the antibacterial experiment of Bifidobacterium lactis subsp. NCU-42 in the embodiments of the present invention are shown.

[0024] Figure 7 This is a statistical chart of the diameter of the inhibition zone of Bifidobacterium lactis subsp. NCU-42 in the embodiments of the present invention.

[0025] Figure 8 The results of the hemolytic activity test of Bifidobacterium lactis subsp. NCU-42 in this embodiment of the invention are shown.

[0026] Figure 9 These are photographs of thyroid tissue from various groups of mice in the embodiments of this invention.

[0027] Figure 10 The graph shows the total weight of the thyroid gland and trachea of ​​mice in each group in this invention embodiment, where * indicates P<0.05, ** indicates P<0.01, and *** indicates P<0.001.

[0028] Figure 11 The results of HE staining of the thyroid glands of mice in each group in the embodiments of the present invention are shown.

[0029] Figure 12 The results of thyroid hormone FT3, FT4, TSH and antibody TRAb detection in each group of mice in the embodiments of the present invention are shown.

[0030] Figure 13 CD4 in each group of mice in the embodiments of the present invention + A statistical chart showing the percentages of Treg cells and Th9 cells in T cells, as well as the ratio of Th9 cells to Treg cells in peripheral blood.

[0031] Figure 14 The CCR9 cells in each group of mouse Th9 cells in the embodiments of the present invention + Percentage of cells and CCR9 in Treg cells + Percentage chart of cells.

[0032] Figure 15 CD4 in each group of mice in the embodiments of the present invention + CCR9 in T cells + Percentage of Treg cells and CD4 + CCR9 in T cells + Percentage of Th9 cells.

[0033] Figure 16 The CCR9 levels in the peripheral blood of mice in each group in this embodiment of the invention. + Th9 cells and CCR9 + The ratio of Treg cells.

[0034] Figure 17 This invention relates to Graves' disease mouse Th9 cells and cytokines IL-9 and CCR9. + Th9 cells and cytokine IL-9, Treg cells and cytokine IL-10, CCR9 + Correlation analysis results of Treg cells and cytokine IL-10. Detailed Implementation

[0035] The following will provide a clear and complete description of the concept and technical effects of the present invention in conjunction with the embodiments and accompanying drawings, so as to fully illustrate the purpose, solution and effects of the present invention.

[0036] Isolation and identification of Bifidobacterium animalis subsp. lactis NCU-42:

[0037] Bacteria were isolated and cultured from fecal samples of healthy volunteers and Graves' disease patients from the Second Affiliated Hospital of Nanchang University, Jiangxi Province. Fecal samples from healthy volunteers contained high abundances of common probiotics such as Bifidobacterium. Different concentrations of bacterial suspensions were prepared using a serial dilution method, and samples of appropriate dilutions were plated (30 μL / plate) and incubated simultaneously in aerobic and anaerobic culture systems for 24-48 hours. Based on colony morphological characteristics (including but not limited to morphological parameters, apparent color, and microstructure), 20-40 typical single colonies were screened and transferred to liquid culture medium for two rounds of activation and proliferation over 24-48 hours. Genomic DNA was extracted from the purified strains, and 16S rDNA sequence amplification and sequencing were performed by Shanghai Sangon Biotech Co., Ltd. The sequencing results are shown in SEQ ID No. 1. Comparative analysis was performed in the NCBI database, and phylogenetic tree construction was used to accurately identify the bacterial species, successfully isolating and culturing *Bifidobacterium animalis* subsp. *lactamella*. Bifidobacterium animalis subsp. lactisIt was named *Bifidobacterium animalis* subsp. *lactobacter* NCU-42 and deposited at the China General Microbiological Culture Collection Center (CGMCC No. 34254). The phylogenetic tree of *Bifidobacterium animalis* subsp. *lactobacter* NCU-42 is as follows: Figure 1 As shown.

[0038] In vitro prebiotic evaluation of Bifidobacterium animalis subsp. lactis NCU-42:

[0039] Bifidobacterium animalis subsp. lactis NCU-42 was inoculated into 100 mL of BSM liquid medium and cultured anaerobically at 37°C. Its optical density was measured at 600 nm at 0 h, 4 h, 8 h, 12 h, 16 h, 20 h, 24 h, 28 h, 32 h, 36 h, 40 h, 44 h, and 48 h. A growth curve was plotted with time on the x-axis and optical density on the y-axis. Figure 2 As shown, the strain enters the stationary phase after about 40 hours of culture and the logarithmic growth phase after about 12-24 hours, indicating that Bifidobacterium animalis subsp. lactis NCU-42 has good growth performance.

[0040] Bifidobacterium animalis subsp. lactis NCU-42 was inoculated into BSM liquid medium and subjected to two activation subcultures at 37°C, each subculture lasting 48 hours. The activated bacterial suspension was then serially diluted: 100 μL of the bacterial suspension was diluted 10 times with sterile PBS buffer. 1 10 3 10 5 The culture was serially diluted, then centrifuged at 4000 rpm for 3 minutes to remove the supernatant. The treated bacterial culture was resuspended in PBS buffer at different pH values ​​(2.0, 3.0, 4.0, 5.0, 7.0) and incubated anaerobically at 37°C for 4 hours. After acid treatment, the bacterial suspension was thoroughly resuspended, and 10 μL of the suspension was spotted onto the surface of BSM solid medium. The culture was then incubated anaerobically at 37°C for another 48 hours before colony counting analysis. Figure 3 As shown, this strain grows stably in an acidic environment with pH 3-7, indicating that it has strong acid resistance.

[0041] BSM basal medium was prepared, and experimental groups were prepared by adding 0.1%, 0.2%, and 0.3% bovine bile salts, respectively. A blank control group without added bile salts was also set up. The activated bacterial strains were inoculated into each medium and incubated in a 37°C anaerobic incubator for 48 hours. After incubation, 10 μL of bacterial suspension was pipetted onto the surface of BSM solid medium using a micropipette. The culture dishes were then further anaerobically cultured under the same conditions for another 48 hours. The surviving cells in each experimental group were quantitatively analyzed using the colony forming unit (CFU) method, and the colony growth and survival rate data under different bile salt concentrations were systematically recorded. Figure 4 As shown, this strain exhibited significant growth (colony count ≥10) in a medium containing 0.3% ox bile salts. 6 The CFU / mL concentration indicates that the strain has a strong tolerance to bile salts.

[0042] The DPPH free radical scavenging rate of *Bifidobacterium lactis* subsp. *lactobiformis* NCU-42 was determined by DPPH free radical scavenging assay; the hydroxyl free radical scavenging rate of *Bifidobacterium lactis* NCU-42 was determined by salicylic acid method; the superoxide free radical scavenging rate of *Bifidobacterium lactis* NCU-42 was determined by pyrogallol method; and the Fe2+ of *Bifidobacterium lactis* NCU-42 was determined by o-phenanthroline spectrophotometry. 2+ The chelating ability was determined by spectrophotometry using potassium ferricyanide as the oxidant to measure the total reducing power of *Bifidobacterium lactis* subsp. *lactobiflorum* NCU-42. For example... Figure 5 As shown, Bifidobacterium animalis subsp. lactis NCU-42 inhibits DPPH free radicals and hydroxyl radicals (OH radicals). - ) and superoxide radicals (O 2- The scavenging abilities of Fe were 96.5%, 62.4%, and 66.2%, respectively. 2+ The chelating ability was 36.3%, and the total reducing power was measured at OD. 700 It is approximately 2.38.

[0043] Bifidobacterium animalis subsp. lactis NCU-42 was inoculated into BSM liquid medium and cultured in an anaerobic incubator at 37°C for 48 h. After centrifugation at 6000 rpm for 10 min, the supernatant was collected. Escherichia coli O157:H7, Salmonella enteritidis, Shigella fexneri, Salmonella typhimurium, Candida albicans, Listeria monocytogenes, β-hemolytic streptococci, and Staphylococcus aureus were revived and inoculated into their respective liquid media, and cultured in a shaker at 37°C for 36 h. The suspensions of each pathogenic bacterium were then spread onto the surface of LB solid medium using the spread plating method, with an inoculation volume of 30 μL per bacterium. Four sterile Oxford cups (Φ=6 mm) were then placed equidistantly on the culture medium surface. 250 μL of centrifuged *Bifidobacterium animalis* subsp. *lactospirum* NCU-42 fermentation supernatant was precisely added to two of the Oxford cups as the experimental group. An equal volume of sterile culture medium was added to the other two cups as a blank control. Each group was tested in duplicate. The mixture was incubated at 37℃, and the size of the inhibition zone was observed every 2 hours. The diameter of the inhibition zone was measured after 8 hours. Figure 6 and Figure 7 As shown, Bifidobacterium animalis subsp. lactis NCU-42 has a good inhibitory effect on Escherichia coli, Salmonella enteritidis, Flexnerella flexneri, Typhoid fever, Candida albicans, Listeria monocytogenes, beta-hemolytic bacteria, Streptococcus, and Staphylococcus aureus. The inhibition zone is greater than 10 mm, indicating that this strain has strong antibacterial properties.

[0044] After subculturing and activating *Bifidobacterium animalis* in BSM liquid medium, the colonies were streaked onto sheep blood agar plates and incubated at 37°C for 48 hours. The presence of hemolytic zones around the colonies was then observed. *Staphylococcus hminis* was used as a positive control to study the hemolytic activity of *Bifidobacterium animalis* subsp. *lactodi* NCU-42. Figure 8 As shown, Bifidobacterium animalis subsp. lactis NCU-42 does not undergo hemolysis.

[0045] The role of Bifidobacterium animalis subsp. lactis NCU-42 in Graves' disease:

[0046] Thirty-six 6-8 week old female BALB / c mice were randomly divided into groups after one week of acclimatization:

[0047] Group C: Mice were fed normally without any treatment (n=12).

[0048] Group M: Mice were immunized with recombinant adenovirus Ad-TSHR289 via the quadriceps femoris muscle in the thigh for modeling. Viral suspension was prepared with 25 μL of sterile diluent, and the single injection dose was 1 × 10⁻⁶. 9 Virus particles (VP) were administered intramuscularly using a microsyringe. The entire immunization program was performed three times, with each intervention spaced 21 days apart, to establish a stable animal model. After the third immunization, animals were simultaneously treated with PBS via gavage for 4 weeks (n=12).

[0049] Group Bif: Similar to Group M, a Graves' disease model was established. After the third immunization, animal Bifidobacterium lactis subsp. NCU-42 (1×10⁻⁶) was administered simultaneously. 9 CFU / mL) gavage treatment for 4 weeks (n=12).

[0050] The effects of Bifidobacterium animalis on Graves disease mice were evaluated by observing the general condition, thyroid size and morphology of the mice, and by detecting thyroid function, antibodies, inflammatory factors and immune cells.

[0051] like Figure 9 and Figure 10 As shown, compared to group C, mice in group M exhibited symmetrical diffuse enlargement of the thyroid gland, with a smooth surface, intact capsule, and visible radial vascular proliferation; the texture was homogeneous and elastic, with no obvious nodules or cystic changes; and the thyroid gland weight was significantly increased (p < 0.001). Compared to group M, mice in group Bif showed reduced symmetrical diffuse enlargement and vascular proliferation of the thyroid tissue, and a relatively reduced thyroid gland weight (p < 0.001); the gland surface was smooth, the capsule was intact, the texture was homogeneous and elastic, and no obvious nodules or cystic changes were observed.

[0052] HE staining results of mouse thyroid glands are as follows Figure 11 As shown, the thyroid gland structure of mice in group C was intact, with follicular epithelial cells arranged in a single layer of cuboidal structure, neatly arranged, with uniformly sized nuclei located at the base; the follicular lumen was filled with homogeneous pink colloid (eosin staining), with smooth edges; the follicles were relatively uniform in size, regularly arranged, and evenly spaced; interstitial blood vessels were sparsely distributed, without abnormal congestion or fibrosis; and there was no infiltration of lymphocytes or plasma cells. In group M mice, thyroid follicular epithelial cell proliferation, follicular lumen shrinkage, and decreased colloid volume were observed, along with interstitial angiogenesis and increased infiltration of lymphocytes or plasma cells. These pathological features provide histological evidence for hyperthyroidism in Graves' disease. Compared to group M, mice in group Bif showed reduced thyroid follicular epithelial cell proliferation, relatively larger follicular lumen, relatively increased colloid volume, and reduced angiogenesis and lymphocyte or plasma cell infiltration, but the overall levels did not reach the levels of group C mice.

[0053] like Figure 12As shown, the levels of FT3, FT4, and TRAb in group M mice were significantly higher than those in group C, while the levels of TSH were significantly lower than those in group C. Furthermore, after intervention with Bifidobacterium lactis subsp. NCU-42, the levels of FT3 and FT4 in group Bif mice were significantly lower than those in group M, while the levels of TSH, although they rebounded, showed no significant difference, and the levels of TRAb showed a decreasing trend.

[0054] High-precision flow cytometry was used to analyze peripheral blood CD4 counts in healthy control mice (Group C), model mice (Group M), and mice treated with Bifidobacterium lactis subsp. NCU-42 (Bif group). + T cell subsets were detected. Results are as follows: Figure 13 As shown, Th9 cells (CD4+) in the peripheral blood of mice in group M + IL9 + The proportion of [unspecified group] was significantly higher than that of the healthy control group (5.11±0.48% vs 1.15±0.11%). p <0.0001), while Treg cells (CD4) + CD25 + FoxP3 + The proportion decreased significantly (1.63±0.25% vs 6.57±0.97%). p <0.0001), suggesting an imbalance of Th9 and Treg cells in Graves' disease mice; and after treatment with Bifidobacterium lactis subsp. NCU-42, Th9 cells (CD4+) decreased. + IL9 + The proportion of [specific group] was significantly lower than that of group M (1.28±0.48% vs 5.11±0.48%). p <0.0001), while Treg cells (CD4) + CD25 + FoxP3 + The proportion of [specific group] was significantly higher than that of group M (6.51±0.82% vs 1.63±0.25%). p <0.0001). Notably, the Th9 / Treg cell ratio was significantly upregulated in the M group (3.17±0.29 vs 0.18±0.03). p <0.0001), and the Th9 / Treg cell ratio was significantly lower in the M group after treatment with Bifidobacterium lactis subsp. NCU-42 (0.19±0.01 vs 3.17±0.29). p <0.0001). These results indicate that Bifidobacterium lactis subspecies NCU-42 can improve the Th9 / Treg immune imbalance in Graves disease mice.

[0055] Based on the crucial role of CCR9 in intestinal homing, further analysis was conducted to determine whether Bifidobacterium animalis could improve CCR9. + Th9 cells (CCR9) + CD4 + IL9 + ) and CCR9 + Treg cells (CCR9) + CD4 + CD25 + FoxP3 + Imbalance of immune cells. The result is as follows: Figure 14 As shown, the CCR9 of group M is lower than that of group C. + The proportion of Th9 cells among all Th9 cells was significantly increased (4.56±0.65% vs 2.52±1.12%). p =0.001), while CCR9 + The proportion of Treg cells was significantly reduced (2.55±0.68% vs 5.06±0.88%). p <0.001); and CCR9 after treatment with Bifidobacterium lactis subsp. NCU-42. + The proportion of Th9 cells was significantly lower in the M group (3.41±0.73% vs 4.56±0.65%). p =0.34), while CCR9 + The proportion of Treg cells was significantly higher in the M group than in the M group (4.71±1.15% vs 2.55±0.68%). p =0.001).

[0056] In addition, such as Figure 15 As shown, compared with group M, mice in group Bif had a higher CCR9 level. + Th9 cells account for a significant portion of CD4+. + The proportion of T cells was significantly reduced (0.12±0.03% vs 0.79±0.15%). p <0.0001), while CCR9 + Treg cells account for a significant portion of CD4+. + The proportion of T cells was significantly increased (0.87±0.17% vs 0.14±0.05%). p <0.0001).

[0057] More noteworthy is that, such as Figure 16 As shown, CCR9 after treatment with Bifidobacterium animals + Th9 / CCR9 + The Treg cell ratio was significantly downregulated (0.15±0.05 vs 6.10±2.16). p <0.0001).

[0058] These results indicate that Bifidobacterium lactis subspecies NCU-42 can improve the Th9 / Treg immune imbalance in Graves disease mice.

[0059] Based on the fact that IL-9 and IL-10 are characteristic cytokines of Th9 cells and anti-inflammatory cytokines secreted by Treg cells, respectively, ELISA detection and correlation analysis with immune cells further corroborated the effect of Bifidobacterium on improving immune imbalance in Graves' disease mice. Results are as follows... Figure 17 As shown, the IL-9 concentration in group M mice was significantly higher than that in group C (26.57±4.39 pg / mL vs 11.80±1.08 pg / mL). p <0.0001), while the level of IL-10, which has immunomodulatory effects, was significantly reduced (5.55±0.85 pg / mL vs 19.47±2.62 pg / mL). p <0.0001). Th9 cells were significantly positively correlated with the level of the inflammatory cytokine IL-9 (r = 0.7397, P = 0.0005), and CCR9... + Th9 cells were also significantly positively correlated with the level of the inflammatory cytokine IL-9 (r = 0.611, P = 0.0028), therefore Th9 cells and CCR9... + Th9 cells play a pro-inflammatory role. Treg cells and CCR9 cells also play a role. + Treg cells were significantly positively correlated with serum IL-10 levels (r = 0.6718, P = 0.0029; r = 0.742, P = 0.0006). Intervention with *Bifidobacterium lactis* subsp. *animal* NCU-42 improved this "dual imbalance" of elevated Th9-characteristic cytokines and decreased regulatory cytokines, which was corroborated by flow cytometry results, confirming that *Bifidobacterium lactis* subsp. *animal* NCU-42 can improve the immune status of Graves' disease mice with Th9 overactivation and Treg dysfunction.

[0060] The above description is merely a preferred embodiment of the present invention. The present invention is not limited to the above-described embodiments. Any embodiment that achieves the technical effects of the present invention using the same means should fall within the protection scope of the present invention. Within the protection scope of the present invention, various modifications and variations can be made to the technical solutions and / or implementation methods.

Claims

1. The application of Bifidobacterium animalis subsp. lactis NCU-42 in the preparation of drugs for the prevention or treatment of Graves' disease, characterized in that, The aforementioned Bifidobacterium lactis subspecies NCU-42 has been deposited at the China General Microbiological Culture Collection Center (CGMCC), located at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, on April 17, 2025, with accession number CGMCC No. 34254. This Bifidobacterium lactis subspecies NCU-42 has the effect of improving hyperthyroidism and Th9 / Treg cell immune imbalance.

2. A drug for the prevention or treatment of Graves' disease, characterized in that, This includes Bifidobacterium animalis subsp. lactis NCU-42, which has been deposited at the China General Microbiological Culture Collection Center (CGMCC) at No. 3, Courtyard 1, Beichen West Road, Chaoyang District, Beijing, on April 17, 2025, with accession number CGMCC No. 34254. Bifidobacterium animalis subsp. lactis NCU-42 has the effect of improving hyperthyroidism and Th9 / Treg cell immune imbalance.

3. The medicament for preventing or treating Graves' disease according to claim 2, characterized in that, The above-mentioned medications for the prevention or treatment of Graves' disease are in liquid form.

4. The medicament for preventing or treating Graves' disease according to claim 3, characterized in that, The aforementioned medications for the prevention or treatment of Graves' disease also include pharmaceutically acceptable excipients.

5. The medicament for preventing or treating Graves' disease according to claim 4, characterized in that, The pharmaceutically acceptable excipients mentioned above are water or physiological saline.

6. The medicament for preventing or treating Graves' disease according to claim 3, characterized in that, The concentration of the above-mentioned Bifidobacterium lactis subsp. NCU-42 was 0.1 × 10⁻⁶. 9 CFU / mL - 10 × 10 9 CFU / mL.

7. The medicament for preventing or treating Graves' disease according to claim 6, characterized in that, The concentration of the above-mentioned Bifidobacterium lactis subsp. NCU-42 was 1×10⁻⁶. 9 CFU / mL - 5 × 10 9 CFU / mL.

8. The medicament for preventing or treating Graves' disease according to claim 7, characterized in that, The concentration of the above-mentioned Bifidobacterium lactis subsp. NCU-42 was 1×10⁻⁶. 9 CFU / mL.

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