Application of tea exosome in preparation of medicine for protecting intestinal barrier and treating irritable bowel syndrome

A technology for irritable bowel syndrome and intestinal barrier, applied in the fields of genetic engineering and life sciences, can solve the problem of unclear regulation of tea exosomes on intestinal barrier, and achieve the effect of protecting function

Pending Publication Date: 2022-06-21
成都市第三人民医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It is unclear whether tea exosomes have a modulatory effect on the intestinal barrier and whether tea exosomes have a therapeutic effect on irritable bowel syndrome

Method used

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  • Application of tea exosome in preparation of medicine for protecting intestinal barrier and treating irritable bowel syndrome
  • Application of tea exosome in preparation of medicine for protecting intestinal barrier and treating irritable bowel syndrome
  • Application of tea exosome in preparation of medicine for protecting intestinal barrier and treating irritable bowel syndrome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] In this example, plant exosomes with a vesicle structure were isolated from tea leaves.

[0050] The BCA kit was purchased from Thermo Fisher Scientific in the United States, the transmission electron microscope (JEM-1400 in Japan) was provided by Chengdu Xuhai Rundong Biotechnology Co., Ltd., the nanoparticle tracking analyzer (Particle Metrix) was produced by Meerbusch in Germany, and the ultracentrifuge (L-100XP) is produced by Beckman Company of the United States.

[0051] Freshly picked buds and 1-2 new leaves of Fuding Dabai (abbreviated as FD), Meizhan (abbreviated as MZ) and Laochuancha (abbreviated as LCC) were used as three types of tea for follow-up research. The exosomes in the three kinds of tea leaves were separated by differential centrifugation. The specific steps were as follows: after the tea leaves were washed with phosphate buffered saline (PBS), they were ground into a homogenate in a mortar, centrifuged at 1000 g for 10 min, and the supernatant was...

Embodiment 2

[0055] In this example, the absorption of tea exosomes in intestinal epithelial cells (Caco-2) was determined.

[0056] Caco-2 was purchased from Changsha Aibiwei Biotechnology Co., Ltd., 24-well cell slide (WHB-24-CS-10) was purchased from Shanghai Wohong Biotechnology Co., Ltd., and reagent DiI (CAS41085-99-8) was purchased from the United States AAT Bioquest company, DAPI (BL105A) was purchased from biosharp company, and fluorescence microscope (Leica Microsystems) was produced by German Wetzlar company.

[0057] DiI-labeled exosomes: Prepare DiI powder with DMSO to a solution with a working concentration of 1 mM, filter through a 0.22 μm filter membrane, and store in the dark. Take 30 μg of exosomes and incubate with 1 μM DiI in the dark for 30 min, and resuspend in PBS. Centrifuge at 100,000g for 2h to remove excess DiI dye.

[0058] DiI-labeled exosomes were co-cultured with cells: Caco-2 cells were inoculated onto cell slides at a concentration of 1×10 4 After 24 hou...

Embodiment 3

[0063] This example investigates the effect of tea exosomes on the barrier function of an in vitro intestinal barrier model.

[0064] Intestinal epithelial cells (CaCo-2) used in the experiment were purchased from Changsha Aibiwei Biotechnology Co., Ltd.; lipopolysaccharide (LPS) was purchased from Sigma-Aldrich (Shanghai) Trading Co., Ltd. 12-well transwell plates (pore size 0.4 μm, 3460), 96-well and 6-well cell culture plates were purchased from Corning Company. Total RNA extraction kit (MiPure Cell / Tissue miRNA Kit, RC201), reverse transcription kit ( III RT SuperMix for qPCR, R323-01) and qPCR kit (ChamQ Universal SYBR qPCR Master Mix, Q711-02 / 03) were purchased from Nanjing Nuoweizan Biotechnology Co., Ltd. ZO-1 antibody (21773-1-AP), Occludin antibody (27260-1-AP) were purchased from proteintech company.

[0065] In vitro intestinal epithelial barrier model based on Transwell chambers: CaCo-2 cells were seeded on 12-well Transwell culture plates at a density of 1×10 ...

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Abstract

The invention provides application of a tea exosome in preparation of a medicine for protecting an intestinal barrier and/or a medicine for treating an irritable bowel syndrome and the medicine for protecting the intestinal barrier and/or treating the irritable bowel syndrome. The tea exosome can be effectively absorbed by intestinal epithelial cells, and the cells are protected from being damaged by lipopolysaccharide by improving the expression of intercellular tight junction proteins ZO-1 and Occludin, so that the barrier function of the intestinal epithelial cells is protected.

Description

technical field [0001] This application relates to the fields of genetic engineering and life sciences, specifically, the application of tea exosomes in the preparation of drugs for protecting the intestinal barrier and treating irritable bowel syndrome. Background technique [0002] The gut barrier maintains homeostasis by preventing the deleterious movement of antigenic molecules and microorganisms from the lumen of the gastrointestinal tract while allowing digestion products and water to enter the body, including physical barriers provided by interepithelial tight junctions, secretory barriers (including antimicrobial peptides, mucus and fluid) and the immune barrier (including cells and molecules of the innate and adaptive immune systems). [0003] Irritable bowel syndrome (IBS) is a common functional bowel disorder characterized by chronic abdominal pain or discomfort and changes in bowel habits. IBS is a common disorder affecting 9-23% of the general population, 80% o...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K36/82A61P1/00
CPCA61K36/82A61P1/00
Inventor 龚前园郭元彪何旭孙悦珊刘蕾张彤彤
Owner 成都市第三人民医院
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