Application of osteopontin as a target molecule in regulating intestinal flora colonization

A technology for regulating intestinal flora and osteopontin, which is applied in the fields of analytical materials, biomaterial analysis, microbial measurement/inspection, etc., to achieve the effect of enhancing adhesion and improving expression

Active Publication Date: 2022-02-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, OPN is a protein secreted by mammalian cells, which is obviously different from the above-mentioned substances that can significantly regulate the intestinal flora of the body. Whether it can be used as a target to regulate the colonization of intestinal flora It is to improve the colonization of probiotics in the intestine and the specific mechanism, and there is no relevant research report so far

Method used

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  • Application of osteopontin as a target molecule in regulating intestinal flora colonization
  • Application of osteopontin as a target molecule in regulating intestinal flora colonization
  • Application of osteopontin as a target molecule in regulating intestinal flora colonization

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1: OPN can affect the colonization of bacterial flora in the intestinal tract of mice

[0024] Eighteen 4-week-old C57BL / 6 mice were selected and fed under the same diet for 24 weeks to determine the content of OPN in the plasma of the mice, and 16S rRNA sequencing was performed on the ileocecal mucosal flora of the mice. Correlation analysis was carried out between the level of OPN in mouse plasma and the abundance of flora, and the results were as follows: figure 1 As shown, the abundance of bacteria such as Olsenella, Bifidobacterium, Turicibacter, Barnesiella, and Lactobacillus was negatively correlated with the OPN level in plasma, The abundance of Mucispirillum, Bacteroides, Alistipes, Dorea, Desulfovibrio and other flora was positively correlated with the level of OPN in plasma.

[0025] Further, 12 4-week-old wild-type and OPN knockout C57BL / 6 mice were respectively selected, and after 24 weeks of feeding under the same dietary conditions, 16s rRNA sequ...

Embodiment 2

[0028] Example 2: OPN can affect the adhesion of bacteria to intestinal epithelial cells

[0029] Intestinal epithelial cells HT-29 were selected for co-culture with Lactobacillus. First, HT-29 cells were cultured in RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin / streptomycin at 37°C, CO 2 The concentration is 8%, and cultured in a carbon dioxide incubator until the cell density is about 10 6 pcs / hole. Lactobacilli were inoculated into LB medium without antibiotics and cultured at 37°C, and counted when they grew to the logarithmic growth phase. Add lactobacillus into the culture medium containing HT-29 cells according to the quantity ratio of 100:1, and then add OPN antibody, OPN (recombinant OPN) and control protein BSA at a final concentration of 1 μg / mL respectively, and use them after co-cultivation for 2 hours Crystal violet stains bacteria and cells and observes them through an oil lens. The result is as image 3 As shown, it can be seen that ...

Embodiment 3

[0031] Example 3: OPN inhibits the expression of cell-associated adhesion factors by binding to cell surface receptors or with the assistance of the Notch signaling pathway

[0032] Intestinal epithelial cells HT-29 were cultured in RPMI-1640 medium containing 10% fetal bovine serum and 1% penicillin / streptomycin at 37°C, CO 2 The concentration is 8%, and cultured in a carbon dioxide incubator until the cell area is about 60% of the orifice plate area. Add OPN cell surface receptor αv, α4 antibody, and OPN antibody (for inhibiting the combination of OPN and its receptor) at a final concentration of 10 μg / mL to the above-mentioned culture medium containing HT-29 cells, or add DAPT (for inhibiting Notch signaling pathway), the final concentrations were 20 μmol / L and 40 μmol / L, respectively. After 2 hours of culture, OPN was added to the culture medium, and the cells and culture medium were collected after continuing to culture for 24 hours, and the transcription level or protei...

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Abstract

The invention discloses the application of osteopontin as a target molecule in regulating intestinal flora colonization; specifically: inhibiting the expression of OPN gene or blocking the combination of OPN and cell receptors, thereby increasing the number of probiotics in the intestinal tract; blocking OPN Binding to cellular receptors or inhibiting Notch signaling can enhance bacterial adhesion to intestinal epithelial cells. The present invention confirms that osteopontin affects the adhesion between cells and bacteria by inhibiting the expression of cell adhesion factors, thereby inhibiting the colonization of bacteria in the intestinal tract.

Description

technical field [0001] The invention relates to an application of osteopontin as a target molecule in regulating colonization of intestinal flora. Background technique [0002] There are a large number of bacteria in the human intestinal tract, the number exceeds 1000 trillion. These intestinal flora participate in the host's nutrient absorption, energy metabolism, immune regulation and other physiological processes, forming a micro-ecosystem with the host. At the same time, the host's health status, life and eating habits also affect the proportion of different bacteria in the intestinal flora. In recent years, many studies have shown that intestinal flora is closely related to the occurrence and development of various human diseases such as diabetes, hypertension, hyperlipidemia, cancer, and nervous system diseases. The metabolites produced by different bacteria in the human gut during their growth and proliferation are considered to be a key factor affecting the health ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/68C12Q1/689C12Q1/6869C12Q1/6851C12Q1/04
CPCG01N33/68C12Q1/689C12Q1/6869C12Q1/6851G01N2333/4728
Inventor 刁宏燕王凯航曾萍章旭君陈佳宁毕珂凡
Owner ZHEJIANG UNIV
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