Lactobacillus delbrueckii capable of killing cervical epithelial cells or cervical cancer cells infected by HPV (human papillomavirus) and application of lactobacillus delbrueckii

Abstract

The invention belongs to the technical field of biological medicines, and particularly relates to lactobacillus delbrueckii with a killing effect on HPV (human papillomavirus) infected cervical epithelial cells or cervical cancer cells and application of the lactobacillus delbrueckii. The classification name of the lactobacillus delbrueckii is Lactobacillus delbrueckii, the preservation number is CGMCC No.32049, the preservation time is September 24, 2024, the preservation place is China General Microbiological Culture Collection Center (CGMCC), and the address of the preservation unit is Institute of Microbiology, Chinese Academy of Sciences, No.3, No.1 Yard, West Beichen Road, Chaoyang District, Beijing. The lactobacillus delbrueckii disclosed by the invention is expected to become a treatment scheme for treating HPV infection, cervical intraepithelial neoplasia and even cervical cancer, and has a relatively high clinical application value and a wide market prospect.

Description

Technical Field

[0001] The present invention belongs to the field of biomedicine technology, and in particular relates to a Lactobacillus delbrueckii having a killing effect on HPV-infected cervical epithelial cells or cervical cancer cells and a use thereof. Background Art

[0002] Lactobacilli play a crucial role in maintaining a healthy vaginal ecosystem, significantly influencing metabolic and immune regulation in the lower reproductive tract. Lactobacilli and their metabolites offer protection against bacterial vaginosis, aerobic vaginitis, viruses, fungi, and protozoa. Therefore, vaginal Lactobacilli homeostasis is a crucial component of vaginal microecological homeostasis. Inflammatory responses caused by various infections contribute to the reactivation of latent HPV, disrupting the balance of the vaginal microecological ecosystem and leading to decreased local vaginal immune function, facilitating HPV invasion. Various vaginal infections may also affect the secretion of certain cytokines in the cervical epithelium, disrupting the immune balance of cervical tissue.

[0003] It usually takes 10-30 years from HPV infection to invasive cancer. 90% of infections can be cleared by the immune system within 2 years, and only 10% develop into persistent infection. Immunosuppression (such as HIV infection) significantly increases the risk of persistent infection. If persistent infection exists, cervical intraepithelial neoplasia (CIN) will occur later, and then develop into invasive cancer. HPV vaccination (primary prevention), regular screening (TCT+HPV testing), and treatment of precancerous lesions constitute the three pillars of eliminating cervical cancer.

[0004] Although a variety of drugs (such as antivirals and immunomodulators) are used to treat HPV infection, drug treatment for HPV-related precancerous lesions remains under investigation, and effective treatment options are yet to be developed. Long-term or frequent use of certain antiviral drugs (such as interferon) can lead to viral gene mutations, resulting in drug resistance. Chemotherapeutic drugs (such as cisplatin) can also develop resistance in cervical cancer, which is related to epigenetic changes and regulation of signaling pathways. In Western medicine, drug resistance can lead to poor treatment outcomes, necessitating timely adjustments to treatment plans. While immunotherapy (such as PD-1 / PD-L1 inhibitors) has shown potential in the treatment of cervical cancer, some patients do not benefit due to a dysregulated immune microenvironment or decreased T cell function. Currently, surgical resection is the main treatment for cervical precancerous lesions, but surgery cannot completely eliminate the HPV virus and carries the risks of pain, bleeding, and infection. It can also lead to cervical shortening, scarring, and contracture, potentially impacting fertility. Furthermore, there is a risk of recurrence after surgical treatment.

[0005] Lactobacilli in the vagina produce substances such as lactic acid and hydrogen peroxide, which lower the vaginal pH, inhibit the growth and reproduction of harmful bacteria, and maintain the vaginal microecological balance. Furthermore, Lactobacilli bind to vaginal epithelial cells to form a biofilm, preventing the adhesion and invasion of pathogens, thereby enhancing the vagina's self-cleaning function and local immune defenses. Lactobacillus preparations are now widely accepted for the treatment of vaginitis. For conditions such as bacterial vaginosis, Trichomonas vaginitis, and vulvovaginal candidiasis, vaginal Lactobacilli can help restore normal vaginal flora after conventional antimicrobial treatment, reducing the recurrence rate of vaginitis.

[0006] However, there are currently no reports on the use of Lactobacillus delbrueckii to prevent HPV infection and treat cervical cancer. Summary of the Invention

[0007] Persistent infection with high-risk HPV is a major cause of cervical intraepithelial neoplasia (CIN) and cervical cancer. To address these shortcomings and deficiencies in the existing technology, the inventors have discovered a novel strain of Lactobacillus delbrueckii that has a targeted killing effect on HPV-infected cervical epithelial cells or cervical cancer cells. This strain of Lactobacillus delbrueckii with this efficacy is expected to become a treatment option for HPV infection, cervical intraepithelial neoplasia, and even cervical cancer.

[0008] In order to achieve the purpose of the present invention, the present invention adopts the following technical solutions:

[0009] In the first aspect, the present invention provides a Lactobacillus delbrueckii, the classification name of the Lactobacillus delbrueckii, the preservation number is CGMCC No.32049, the preservation time is September 24, 2024, the preservation location is the General Microbiology Center of the China Culture Collection Administration (CGMCC), and the preservation unit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0010] As an optional mode, in the above Lactobacillus delbrueckii, the Lactobacillus delbrueckii has a killing effect on HPV-infected cervical epithelial cells or cervical cancer cells, and promotes apoptosis of HPV-infected cervical epithelial cells or cervical cancer cells.

[0011] In a second aspect, the present invention provides a Lactobacillus delbrueckii supernatant, which is prepared from the Lactobacillus delbrueckii described in the first aspect.

[0012] As an optional mode, in the above-mentioned Lactobacillus delbrueckii supernatant, the preparation method of the Lactobacillus delbrueckii supernatant comprises the following steps:

[0013] The Lactobacillus delbrueckii was revived in MRS broth, and the inoculated broth was cultured in an anaerobic or microaerobic incubator at 37°C for 24 to 48 hours. The centrifuge tube was centrifuged at 2000-3000g for 5 to 10 minutes to allow the cells to form a pellet at the bottom of the tube. After centrifugation, the supernatant was carefully aspirated without disturbing the cell pellet.

[0014] Preferably, the method for preparing the Lactobacillus delbrueckii supernatant comprises the following steps:

[0015] To resuscitate the L. delbrueckii in MRS broth, first prepare the broth according to a standard sterile recipe and set the water bath temperature to 37°C. If using a frozen culture, remove the vial from liquid nitrogen and quickly thaw it in a water bath while gently stirring until completely thawed. Then, sterilize the outside of the vial with 70% ethanol. Transfer the thawed culture to a sterile conical tube containing 10 mL of prewarmed MRS broth. Incubate the inoculated broth in an anaerobic or microaerobic incubator at 37°C for 24 to 48 hours. Centrifuge the tube at 2000-3000g for 5 to 10 minutes to allow the cells to form a pellet at the bottom of the tube. After centrifugation, carefully aspirate the supernatant without disturbing the cell pellet.

[0016] As an optional manner, in the above-mentioned Lactobacillus delbrueckii supernatant, the Lactobacillus delbrueckii supernatant has a killing effect on HPV-infected cervical epithelial cells or cervical cancer cells, and promotes the apoptosis of HPV-infected cervical epithelial cells or cervical cancer cells.

[0017] In a third aspect, the present invention provides use of the Lactobacillus delbrueckii described in the first aspect or the supernatant of the Lactobacillus delbrueckii described in the second aspect in the preparation of a medicament for preventing or treating HPV infection.

[0018] In a fourth aspect, the present invention provides use of the Lactobacillus delbrueckii described in the first aspect or the Lactobacillus delbrueckii supernatant described in the second aspect in the preparation of a medicament for preventing or treating cervical intraepithelial neoplasia or cervical cancer.

[0019] As an option, in the above use, the medicine further comprises other chemotherapeutic drugs.

[0020] As an option, in the above use, the other chemotherapeutic drug is cisplatin or paclitaxel.

[0021] Preferably, the chemotherapeutic drug is cisplatin, and the Lactobacillus delbrueckii described in the first aspect or the supernatant of Lactobacillus delbrueckii described in the second aspect can enhance the therapeutic effect of cisplatin.

[0022] As an optional mode, in the above use, the dosage form of the drug is an external dosage form. Preferably, the external dosage form is selected from a solution, emulsion, cream, ointment, gel or spray.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The inventors have discovered a new type of Lactobacillus delbrueckii (CGMCC No. 32049) that has a targeted killing effect on HPV-infected cervical epithelial cells or cervical cancer cells. This Lactobacillus delbrueckii with this efficacy is expected to become a treatment option for HPV infection, cervical intraepithelial neoplasia, and even cervical cancer, with high clinical application value and broad market prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0026] Figure 1 : Gram staining results after co-culture of HeLa cells with different lactobacilli. Among them: (A) Lactobacillus crispatus, (B) Lactobacillus johnsonii, (C) Lactobacillus jensenii, (D) Lactobacillus garceri, (E) Lactobacillus fragilis, (F) Lactobacillus vaginalis, (G) Lactobacillus helveticus, (H) Lactobacillus delbrueckii, 20x.

[0027] Figure 2 Microscopic imaging of live HeLa cells co-cultured with Lactobacillus delbrueckii for 24 hours, 100x. Within the red circle, numerous Lactobacilli are seen crawling around the HeLa cells, and the HeLa cells are clearly deformed and on the verge of death.

[0028] Figure 3 Real-time observation of cellular changes in HeLa cells co-cultured with Lactobacillus delbrueckii from 0 to 24 hours, 60x. Blue circles indicate floating HeLa cells that have emerged from the bottom of the culture plate.

[0029] Figure 4 : RT-qPCR detection of HeLa apoptosis-related gene expression. *p<0.01, two-way ANOVA.

[0030] Figure 5: Annexin-V / PI cell apoptosis experiments confirmed that the combination of Lactobacillus delbrueckii supernatant and cisplatin significantly promoted late apoptosis of HeLa cells. Figure A shows the results of flow cytometry analysis showing that in the control group not treated with supernatant, the proportion of annexin V-positive cells was only 1.4%, indicating that the baseline level of cell apoptosis in HeLa cells not treated with supernatant was low. In the experimental group treated with 500μg / mL Lactobacillus delbrueckii supernatant, the proportion of annexin V-positive cells increased significantly, rising to 25.6%. Figure B evaluated the effect of Lactobacillus delbrueckii supernatant combined with chemotherapy drugs (cisplatin / paclitaxel) on HeLa cell apoptosis by Annexin V / PI staining and flow cytometry. The proportion of PI-positive cells in the control group (10nM cisplatin) was 11.2%, while that in the experimental group (500μg / ml Lactobacillus supernatant + 10nM cisplatin) increased to 66.2%. These results suggest that the combined use of Lactobacillus supernatant and cisplatin can significantly promote the late apoptosis of HeLa cells induced by cisplatin.

[0031] Figure 6 : Cervical cancer cells treated with Lactobacillus delbrueckii supernatant and cisplatin were able to inhibit tumor growth in mice with cervical cancer transplants after inoculation into mice. Figure A is a mouse model construction plan that lasts for 28 days, with drug treatment starting on the treatment day (the 10th day after inoculation of HeLa cells). The study was divided into four groups: a control group (no treatment), Lactobacillus delbrueckii plus cisplatin, Lactobacillus delbrueckii alone, and cisplatin alone. Figure B shows that the tumor size was measured every day for 18 days after the start of treatment. Figure C shows that after the mice were killed on the 28th day, the weight of the tumors in the three study groups was significantly reduced compared with the control group, and the most obvious inhibition of tumor growth was in the Lactobacillus delbrueckii plus cisplatin group. DETAILED DESCRIPTION

[0032] The present invention will be further described below with reference to specific embodiments. It should be understood that the specific embodiments described herein are only used to illustrate the present invention and are not intended to limit the scope of the present invention.

[0033] If no specific techniques or conditions are specified in the examples, the experiments were carried out according to the techniques or conditions described in the literature in the field or according to the product instructions. If no manufacturer is specified for the reagents or instruments used, they are all conventional products that can be purchased through regular channels.

[0034] The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are commercially available products unless otherwise specified.

[0035] Example:

[0036] Example 1: Co-culture experiment of various lactic acid bacteria and HeLa cells

[0037] Co-culturing lactic acid bacteria with HeLa cells is an experimental method used to study the interaction between probiotics and cancer cells.

[0038] HeLa cells were cultured in DMEM containing 10% FBS and 1% penicillin-streptomycin at 37°C and 5% CO2 until they reached 70-80% confluence, while eight lactobacilli strains were cultured in MRS broth under anaerobic or microaerobic conditions at the same temperature for 24-48 hours until significant growth was observed; subsequently, HeLa cells were harvested with 0.25% trypsin-EDTA and resuspended to approximately 1 × 10 5 to 1×10 6 The cells were grown at a density of 10 cells / mL, after which the lactobacillus strains were added at various multiplicity of infection (MOI) ranging from 1:1 to 10:1 and incubated again at 37°C with 5% CO2 for 24-48 hours. Gram-stained smears were then prepared from samples collected after incubation, air-dried and heat-fixed before the Gram-staining process, which included sequential staining with crystal violet, iodine solution, ethanol and safranin for decolorization, followed by rinsing and air-drying. The stained slides were then examined under a light microscope, where Gram-positive lactobacillus cells appeared purple and Gram-negative cells appeared pink, allowing for the assessment of cell viability. Intact and purple-stained lactobacilli indicated viability in the presence of HeLa cells. Reduced viability, including poor purple staining or an increase in pink cells, indicated negative interactions during co-culture.

[0039] The eight Lactobacillus species selected for study include a novel strain of Lactobacillus delbrueckii, newly isolated and identified by the inventors from vaginal secretions of healthy women. Its classification name is Lactobacillus delbrueckii, with a deposit number of CGMCC No. 32049 and a deposit date of September 24, 2024, at the General Microbiology Center of the China Culture Collection Administration (CGMCC). The depository address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

[0040] In addition to the Delbrück strain of Lactobacillus, there are seven other standard strains: Lactobacillus crispatus (DSM 20584), Lactobacillus johnsonii (ATCC 33200), Lactobacillus jensenii (DSM 20557), Lactobacillus gasseri (ATCC 33323), Lactobacillus fragilis (BAA3226), Lactobacillus vaginalis (ATCC 49540), and Lactobacillus helveticus (DSM 20075). These eight types of lactobacilli were co-cultured with HeLa cells for 12 hours and then stained using the Gram staining method described above. The results are shown in Figure 2. Figure 1As shown in the figure, under a 20x microscope, the number of HeLa cells decreased when co-cultured with vaginal Lactobacillus (ATCC 49540), Lactobacillus fragilis (BAA 3226), and Lactobacillus delbrueckii (CGMCC No. 32049), with Lactobacillus delbrueckii (CGMCC No. 32049) showing a significantly better effect than vaginal Lactobacillus and Lactobacillus fragilis. Co-culture with the other strains had no effect on the number of HeLa cells.

[0041] Example 2: Live cell microscopy imaging of co-culture of Lactobacillus delbrueckii with HeLa cells or HCerEpiC cells result

[0042] The preparation of cell culture involves culturing HeLa cells in DMEM supplemented with 10% FBS and 1% penicillin-streptomycin in a CO2 incubator until they reach 70-80% confluence, while growing Lactobacillus delbrueckii (CGMCC No. 32049) in MRS broth under anaerobic or microaerobic conditions at 37°C for 24-48 hours until significant growth is achieved; for the co-culture setup, HeLa cells are harvested using trypsin-EDTA and resuspended in DMEM to reach a confluence of approximately 1×10 5 to 1×10 6 Delbrueckii was added to a density of 10 cells / mL, followed by the addition of Lactobacillus delbrueckii at an appropriate multiplicity of infection (MOI) of 1:1 to 10:1, and the co-culture was incubated at 37°C in a humidified atmosphere with 5% CO2 for 24 hours; at the end of the incubation period, the co-culture was prepared for microscopy, HeLa cells or Lactobacillus delbrueckii were selectively labeled, and then a small co-culture volume was transferred to a glass-bottom dish or imaging chamber suitable for live cell microscopy; the samples were continuously monitored to observe the interaction between Lactobacillus delbrueckii and HeLa cells.

[0043] The experimental results showed that after 24 hours, the widespread presence of Lactobacillus delbrueckii was observed, making the HeLa cells almost invisible to the naked eye. Figure 2 The HeLa cells shown are identifiable, and most of them have been infiltrated and damaged to the point of death. This suggests that Lactobacillus delbrueckii can induce the death of HeLa cells. Figure 3 As shown, in the real-time observation of cell changes from 0 to 24 hours, it was observed that the number of Lactobacillus delbrueckii gradually increased over time, and HeLa cells fell off and floated from the bottom of the culture plate, indicating that the HeLa cells were almost dead after 24 hours of co-culture.

[0044] In this study, a normal cervical epithelial cell line (HCerEpiC) was used as a positive control to determine whether L. delbrueckii specifically targets cervical cancer cells (HeLa cells) and causes cell death, using the same experimental methods as described above. HCerEpiC 1 (YC246) and HCerEpiC 2 (YC240) are cervical epithelial cell lines derived from normal cervical cells obtained from healthy women attending the Yale University Hospital outpatient gynecology department. Patient consent was obtained during sample collection to ensure the ethical safety of these cell lines for research purposes.

[0045] Normal cervical cells were exposed to the Lactobacillus delbrueckii supernatant prepared using the methods of the present invention. The results showed that, in contrast to the cell death induced in cancer cells, normal cervical cells did not undergo cell death. Notably, normal cervical cells even proliferated in the presence of Lactobacillus delbrueckii, suggesting its potential beneficial or non-toxic effects on healthy cervical epithelium.

[0046] The preparation method of Lactobacillus delbrueckii supernatant of the present invention comprises the following steps:

[0047] To resuscitate the L. delbrueckii in MRS broth, first prepare the broth according to a standard sterile recipe and set the water bath temperature to 37°C. If using a frozen culture, remove the vial from liquid nitrogen and quickly thaw it in a water bath while gently stirring until completely thawed. Then, sterilize the outside of the vial with 70% ethanol. Transfer the thawed culture to a sterile conical tube containing 10 mL of prewarmed MRS broth. Incubate the inoculated broth in an anaerobic or microaerobic incubator at 37°C for 24 to 48 hours. Centrifuge the tube at 2000-3000g for 5 to 10 minutes to allow the cells to form a pellet at the bottom of the tube. After centrifugation, carefully aspirate the supernatant without disturbing the cell pellet.

[0048] Example 3: Lactobacillus delbrueckii acts on apoptosis-related proteins and signaling pathways Caspase and Bcl-2 family Protein promotes apoptosis in HeLa cells

[0049] The concentration of effective protein in the supernatant of Lactobacillus delbrueckii (prepared as in Example 2) was determined by BCA method, and the supernatant was prepared with normal saline to test concentrations of 0 μg / mL, 100 μg / mL, 500 μg / mL and 1000 μg / mL for subsequent studies. Figure 4As shown, in HeLa cells treated with 500 μg / mL Lactobacillus delbrueckii supernatant, the mRNA expression levels of Caspase-3, Caspase-9, and Bax were significantly upregulated, indicating that Lactobacillus delbrueckii activated apoptotic signaling pathways in these cells. The upregulation of Caspase-3 and Caspase-9 suggests enhanced execution of apoptosis. The increased expression of the pro-apoptotic protein Bax further suggests that Lactobacillus delbrueckii promotes apoptosis by shifting the balance toward apoptotic signaling. The significant decrease in Bcl-2 mRNA expression suggests that the cells' anti-apoptotic defenses are weakened, making tumor cells more susceptible to apoptosis. These results suggest that Lactobacillus delbrueckii affects the proliferation of cervical cancer cells by affecting apoptosis-related signaling pathways.

[0050] Example 4: Annexin-V / PI apoptosis assay confirmed that when Lactobacillus delbrueckii supernatant was used in combination with cisplatin Significantly promotes late apoptosis of HeLa cells

[0051] 3 × 10 cells per well in a 6-well plate 4 HeLa cells were inoculated at a density of 10 cells / mL. The cells were allowed to adhere and grow for 24 hours. Following this incubation period, the cells were treated with 500 μg / mL of the prepared Lactobacillus delbrueckii supernatant (prepared as in Example 2) as determined by the BCA assay, as well as with 10 nM cisplatin or 2 nM paclitaxel. Experiments were initiated 12 hours after treatment.

[0052] like Figure 5 As shown in A, the flow cytometry results showed that the control group that did not receive any Lactobacillus delbrueckii supernatant treatment (0 μg / mL) had only 1.4% Annexin V positive cells, indicating that the baseline level of apoptosis in untreated HeLa cells was low. In contrast, the experimental group that received 500 μg / mL Lactobacillus delbrueckii supernatant showed a significant increase in the proportion of Annexin V positive cells, rising to 25.6%. This difference highlights the strong apoptotic effect of Lactobacillus delbrueckii at the tested concentration. Figure 5 As shown in Figure B, the proportion of PI-positive cells in the control group (10 nM cisplatin) was 11.2%, while in the experimental group (500 μg / mL Lactobacillus delbrueckii supernatant + 10 nM cisplatin), it increased to 66.2%. This suggests that the combination of Lactobacillus delbrueckii supernatant and cisplatin significantly promotes cisplatin-induced late apoptosis in HeLa cells. However, this significant effect was not observed when 500 μg / mL Lactobacillus delbrueckii supernatant was combined with 2 nM paclitaxel.

[0053] Example 5: Mouse human cell xenograft (CDX) model shows that Lactobacillus delbrueckii enhances and maintains the therapeutic effect of cisplatin

[0054] The 28-day study used a cell-derived xenograft (CDX) model to demonstrate the differential efficacy of targeted cancer therapy. The protocol is available at Figure 6A, specifically divided into four groups: control group (NT), Lactobacillus delbrueckii plus cisplatin (L.del + Cisplatin), Lactobacillus delbrueckii (L.del) alone, and Cisplatin alone. The cell line used in the study was HeLa cells (ATCC CCL-2), and the mice were 6-week-old female BALB / c nude mice. Cisplatin (Cisplatin, purchased from Promega USA). At the beginning of the study, 6×10 6 100 μL HeLa cell suspension of 100 cells was subcutaneously injected into the flank of anesthetized mice. The size of the tumor was measured with a caliper every day from the time it could be felt, and the tumor volume was calculated using the following formula: Volume = (length × width × depth) / 2. The treatment period started on the 10th day after tumor injection and lasted for 7 days. The treatment methods were: Cisplatin group, each mouse was injected with 0.01 mL of 4 mg / mL cisplatin per day during the treatment period; Lactobacillus delbrueckii group, each mouse tumor cell was injected with 0.01 mL of Lactobacillus delbrueckii supernatant during the treatment period; Lactobacillus delbrueckii plus cisplatin group, the treatment during the treatment period was the sum of the treatments of the cisplatin and Lactobacillus delbrueckii groups. Figure 6 As shown in B, the tumor volume of the groups receiving Lactobacillus delbrueckii, cisplatin, and Lactobacillus delbrueckii plus cisplatin treatment all showed a reduction and sustained maintenance. Among them, the tumor volume reduction effect of the Lactobacillus delbrueckii plus cisplatin group was the most significant, suggesting that the combination of the two may have a synergistic anti-tumor effect. Figure 6 As shown in C, the tumors obtained after euthanasia were measured and weighed. Figure 6 The changes in tumor volume were consistent with those shown in B. Among them, the Lactobacillus delbrueckii plus cisplatin group had the most significant effect on reducing tumor weight, suggesting that the combination of the two may have a synergistic anti-tumor effect.

[0055] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A Lactobacillus delbrueckii, characterized in that: The classification name of the Lactobacillus delbrueckii is (Lactobacillusdelbrueckii), the preservation number is CGMCC No.32049, the preservation time is September 24, 2024, the preservation location is the General Microbiology Center of the China Culture Collection Administration (CGMCC), and the preservation unit address is the Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, Beichen West Road, Chaoyang District, Beijing.

2. Lactobacillus delbrueckii according to claim 1, characterized in that: The Lactobacillus delbrueckii has a killing effect on HPV-infected cervical epithelial cells or cervical cancer cells, and promotes the apoptosis of HPV-infected cervical epithelial cells or cervical cancer cells.

3. A Lactobacillus delbrueckii supernatant, characterized in that: The Lactobacillus delbrueckii supernatant is prepared from the Lactobacillus delbrueckii described in claim 1.

4. The Lactobacillus delbrueckii supernatant according to claim 3, wherein: The preparation method of the lactobacillus delbrueckii supernatant comprises the following steps: The Lactobacillus delbrueckii was revived in MRS broth, and the inoculated broth was cultured in an anaerobic or microaerobic incubator at 37°C for 24 to 48 hours. The centrifuge tube was centrifuged at 2000-3000g for 5 to 10 minutes to allow the cells to form a pellet at the bottom of the tube. After centrifugation, the supernatant was carefully aspirated without disturbing the cell pellet.

5. The Lactobacillus delbrueckii supernatant according to claim 3, wherein: The Lactobacillus delbrueckii supernatant has a killing effect on HPV-infected cervical epithelial cells or cervical cancer cells, and promotes the apoptosis of HPV-infected cervical epithelial cells or cervical cancer cells.

6. Use of the Lactobacillus delbrueckii according to claim 1 or the supernatant of Lactobacillus delbrueckii according to claim 3 in the preparation of a medicament for preventing or treating HPV infection.

7. Use of the Lactobacillus delbrueckii according to claim 1 or the supernatant of Lactobacillus delbrueckii according to claim 3 in the preparation of a medicament for preventing or treating cervical intraepithelial neoplasia or cervical cancer.

8. The use according to claim 6 or claim 7, characterized in that: The drug may also include other chemotherapeutic drugs.

9. The use according to claim 8, characterized in that: The other chemotherapy drug is cisplatin or paclitaxel.

10. The use according to claim 6 or claim 7, characterized in that: The dosage form of the drug is an external dosage form, and the external dosage form is selected from a solution, emulsion, cream, ointment, gel or spray.

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