Search

CN-121975654-A - Lactobacillus paracasei extracellular vesicle with function of regulating iCAF as well as preparation method and application thereof

CN121975654ACN 121975654 ACN121975654 ACN 121975654ACN-121975654-A

Abstract

The invention relates to the technical field of biology, in particular to a lactobacillus paracasei extracellular vesicle with a function of regulating iCAF. The invention provides a lactobacillus paracasei extracellular vesicle with a function of regulating iCAF, which has a spherical structure, has a particle size distribution of 50nm to 200 nm and expresses at least one marker protein of exosome marker proteins CD63, CD9 and TSG 101. The lactobacillus paracasei extracellular vesicles are used for preparing medicines for inhibiting the tumor promotion and drug resistance functions of the fibroblasts related to the inflammatory cancers and preparing medicines for inhibiting the tumor promotion and metastasis functions of the fibroblasts related to the inflammatory cancers.

Inventors

  • XU KE
  • XU HANZHENG
  • LIU HAN
  • YU XIAOBO
  • WANG ZHAOJUN
  • LIU JINLONG

Assignees

  • 上海大学温州研究院
  • 温州市中心医院
  • 苏州上海大学创新中心
  • 苏州申博益维生物医药科技有限公司

Dates

Publication Date
20260505
Application Date
20251225

Claims (10)

  1. 1. The lactobacillus paracasei extracellular vesicle with the function of regulating iCAF is characterized by having a spherical structure, having a particle size distribution of 50 nm to 200 nm and expressing at least one marker protein of exosome marker proteins CD63, CD9 and TSG 101.
  2. 2. Use of an extracellular vesicle of lactobacillus paracasei according to claim 1 for the preparation of a medicament for inhibiting a tumor-promoting drug-resistant function of inflammatory cancer-related fibroblasts.
  3. 3. The use of claim 2, wherein inhibiting inflammatory cancer-associated fibroblast activity comprises down-regulating expression of markers PDGFRA and/or IL-6 that inhibit inflammatory cancer-associated fibroblasts, and/or inhibiting the level of phosphorylation of STAT3 signaling pathway.
  4. 4. Use of an extracellular vesicle of lactobacillus paracasei according to claim 1 for the preparation of a medicament for inhibiting inflammatory cancer related fibroblast tumor metastasis.
  5. 5. The use according to any one of claims 2 or 4, wherein the extracellular vesicles of lactobacillus paracasei mediate an inhibitory effect on iCAF by miR-26a-5 p.
  6. 6. A method for preparing the extracellular vesicles of lactobacillus paracasei having the function of controlling iCAF according to claim 1, comprising the following steps: (1) Inoculating lactobacillus paracasei into MRS liquid culture medium for culture, and collecting culture solution; (2) Centrifuging the culture solution, collecting supernatant, and filtering and sterilizing by using a filter membrane; (3) Ultracentrifugation is carried out on the filtered supernatant, the supernatant is removed, and sediment containing extracellular vesicles of lactobacillus paracasei is obtained; (4) And re-suspending the precipitate in a buffer solution to obtain the lactobacillus paracasei extracellular vesicles.
  7. 7. The method for preparing extracellular vesicles of Lactobacillus paracasei with function of controlling iCAF according to claim 6, wherein in the step (2), the centrifugation is performed at 4 ℃ for 20 minutes under 10000 g, and the pore size of the filter membrane is 0.22 μm.
  8. 8. The method for producing an extracellular vesicle of Lactobacillus paracasei having a function of controlling iCAF according to claim 6, wherein in the step (3), the condition of the ultracentrifugation is 4 ℃ and 150000 g, and the centrifugation is performed for 2 hours.
  9. 9. The method for preparing extracellular vesicles of lactobacillus paracasei having function of controlling iCAF according to claim 6, wherein in step (4), the buffer is PBS buffer.
  10. 10. A pharmaceutical composition comprising the lactobacillus paracasei extracellular vesicles of claim 1 and a pharmaceutically acceptable carrier.

Description

Lactobacillus paracasei extracellular vesicle with function of regulating iCAF as well as preparation method and application thereof Technical Field The invention relates to the technical field of biology, in particular to a lactobacillus paracasei extracellular vesicle with a function of regulating iCAF and a preparation method and application thereof. Background The intestinal microbiota contains pathogenic bacteria (such as Fusobacterium nucleatum and Escherichia coli) and probiotics (such as Clostridium butyricum and Lactobacillus paracasei) for promoting colorectal cancer, and affects colorectal cancer development together with viruses and fungi. The probiotics have the advantages of definite mechanism, wide application, intestinal tract protection, immunoregulation, antibiosis, anti-inflammatory, anti-tumor and the like after years of researches. Probiotics are required to colonize the intestinal tract and are closely related to host metabolism, immunity and intestinal health. Lactobacillus paracasei inhibits tumor proliferation by regulating inflammatory factors, enhancing immune regulation, relieving inflammatory bowel disease, and regulating apoptosis proteins, active oxygen, etc., and extracellular vesicles thereof participate in cell communication. One function that bacteria often ignore is the release of Extracellular Vesicles (EV), a nano-sized vesicle composed of lipid bilayer membranes for the transport of various biomolecules. Emerging studies have found that bacteria can regulate intestinal health by producing Bacterial Extracellular Vesicles (BEVs). BEV is a class of cellular products secreted by gram-negative and positive bacteria. These vesicles are typically 20-400 nanometers in diameter and have a bilayer lipid membrane structure. Under the protection of the membrane, the BEV encapsulates various substances including virulence factors, proteins, nucleic acids and lipids. Bacterial EVs are able to cross epithelial cells, and thus can be found in extracellular spaces and in various biological fluids, and play a critical role in the transport of micrornas (mirnas), messenger RNAs (mrnas) and proteins from parent cells to recipient cells. Based on the above, the invention provides a lactobacillus paracasei extracellular vesicle with function of regulation iCAF, and a preparation method and application thereof. Disclosure of Invention In order to achieve the aim, the invention provides the lactobacillus paracasei extracellular vesicle with the function of regulating iCAF, and the preparation method and the application thereof. In one aspect, the invention provides a lactobacillus paracasei extracellular vesicle with a regulatory iCAF function, which has a spherical structure, has a particle size distribution of 50 nm to 200 nm, and expresses at least one marker protein of exosome marker proteins CD63, CD9 and TSG 101. On the other hand, the invention also provides application of the lactobacillus paracasei extracellular vesicles in preparing medicines for inhibiting inflammatory cancer related fibroblast tumor promotion drug resistance. Further, the inhibiting inflammatory cancer-associated fibroblast activity comprises down-regulating expression of a marker PDGFRA and/or IL-6 that inhibits inflammatory cancer-associated fibroblasts, and/or inhibiting the level of phosphorylation of STAT3 signaling pathway. On the other hand, the invention also provides application of the lactobacillus paracasei extracellular vesicles in preparing medicines for inhibiting inflammatory cancer related fibroblast tumor metastasis function. Further, the Lactobacillus paracasei extracellular vesicles mediate the inhibition of iCAF by miR-26a-5 p. On the other hand, the invention also provides a preparation method of the lactobacillus paracasei extracellular vesicles with function of regulating iCAF, which comprises the following steps: (1) Inoculating lactobacillus paracasei into MRS liquid culture medium for culture, and collecting culture solution; (2) Centrifuging the culture solution, collecting supernatant, and filtering and sterilizing by using a filter membrane; (3) Ultracentrifugation is carried out on the filtered supernatant, the supernatant is removed, and sediment containing extracellular vesicles of lactobacillus paracasei is obtained; (4) And re-suspending the precipitate in a buffer solution to obtain the lactobacillus paracasei extracellular vesicles. Further, in the step (2), the centrifugation condition is that the temperature is 4 ℃,10000 g and the centrifugation is carried out for 20 minutes, and the pore size of the filter membrane is 0.22 mu m. Further, in the step (3), the condition of the ultracentrifugation is 4 ℃ and 150000 g, and the ultracentrifugation is performed for 2 hours. Further, in step (4), the buffer is a PBS buffer. In another aspect, the invention also provides a pharmaceutical composition comprising the lactobacillus paracasei extracellular vesicles and a pharmaceutically