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Application of spermidine in preparing medicine for treating impaired intestinal barrier function

A spermidine and barrier technology is applied in the application field of spermidine in the preparation of drugs for treating impaired intestinal barrier function, and can solve the problem of intestinal bacteria and endotoxin translocation, increased intestinal barrier permeability, and epithelial cells. Problems such as broken valid connections

Inactive Publication Date: 2019-05-21
ZHEJIANG UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, under external stimuli or damage, the normal morphology of intestinal epithelial cells changes, the arrangement is disordered, and the effective connection between epithelial cells is destroyed, resulting in increased intestinal barrier permeability, causing and aggravating intestinal inflammatory response, resulting in intestinal histopathology damage, which in turn causes the translocation of intestinal bacteria and endotoxin and induces systemic infection

Method used

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  • Application of spermidine in preparing medicine for treating impaired intestinal barrier function
  • Application of spermidine in preparing medicine for treating impaired intestinal barrier function
  • Application of spermidine in preparing medicine for treating impaired intestinal barrier function

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] The human colon cancer cell line Caco2 was purchased from the Cell Bank of the Chinese Academy of Sciences in Shanghai, and cultured in a low-glucose medium plus 20% fetal bovine serum and 100 U / mL penicillin and streptomycin in an incubator at 37°C and 5% CO2. Subculture was performed every four days, and the medium was changed every two days.

[0024] LPS solutions with different final concentrations (0, 0.1, 1, 10, 100, 1000 ng / mL) were used for the test, a total of 6 treatments, 6 replicates for each treatment, and one well of cells for each replicate. According to the results of cell permeability changes and cytotoxic activity after 5 days of treatment, the optimal stress dose was determined to be 100ng / mL for follow-up research.

Embodiment 2

[0026] Determination of Optimum Dose of Spermidine

[0027] Caco-2 cells were cultured in vitro, and the culture conditions were the same as in Example 1. After the cells covered the culture dish, the cells were plated in a 6-well plate, and the optimal concentration was explored with different concentrations (0, 50, 100, 200 μM) of spermidine solutions. The experiment was divided into 5 treatments, each treatment had 6 replicates, and each replicate had 1 well of cells. At the same time, the cells were treated with lipopolysaccharide and spermidine solutions of different concentrations, respectively:

[0028] 1) Blank control group: no treatment group;

[0029] 2) LPS stress model group: 100ng / mL LPS treatment;

[0030] 3) Spermidine treatment group: 100ng / mL lipopolysaccharide treatment, plus 0, 50, 100, 200μM spermidine solutions respectively;

[0031] After 5 days, the cells were uniformly collected for subsequent studies.

Embodiment 3

[0033] Detection of expression level of tight junction protein mRNA in cells by reverse transcription-polymerase chain reaction

[0034] Takara RNAiso Plus was used to extract the total RNA of the cells in Example 2, and the RNA concentration and quality were detected. 2 μL of the RNA stock solution was taken to measure the OD value and concentration at 280nm and 260nm. The OD value between 1.8 and 2.0 meets the concentration requirements of reverse transcription for total RNA. Then, configure the reverse transcription reaction system for the detected mRNA according to the instructions of the reverse transcription kit, mix well and incubate at 37°C for 15 minutes, then reverse transcription at 98°C for 5 minutes, and place the prepared cDNA template at -20°C Store in refrigerator until use.

[0035] use Green Real Time PCR Master Mix (Toyobo, China) was used for real-time quantitative polymerase chain reaction analysis, and the reaction system was 10 μL. Primer sequences w...

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Abstract

The invention discloses application of spermidine in preparing medicine for treating the impaired intestinal barrier function. An in vivo and in vitro cell and intestinal permeability increase model is built through lipopolysaccharide stimulation, and the regulation and mechanism of the spermidine on the intestinal barrier function are studied through mutual corroboration.

Description

technical field [0001] The invention relates to the application of a polyamine substance spermidine, in particular to the application of spermidine in the preparation of medicines for treating impaired intestinal barrier function. Background technique [0002] The intestinal tract is an innate barrier to maintain the balance of the intestinal environment and block pathogenic bacteria and toxins. The intestinal barrier includes mechanical barriers, chemical barriers, microbial barriers and immune barriers. The mechanical barrier is also called the physical barrier, and its physiological structure is based on the mucosal epithelium, lamina propria and muscularis mucosae. Intestinal epithelial cells are tightly arranged through cell junctions, and the connections between cells are composed of tight junctions, adherens junctions and desmosomes, which can effectively block the entry of bacteria, viruses and endotoxins. The chemical barrier is composed of mucus, digestive juices ...

Claims

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Application Information

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IPC IPC(8): A61K31/132A61P1/00
Inventor 倪银华马灵燕傅正伟
Owner ZHEJIANG UNIV OF TECH
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