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Application of trefoil factor 2 in preparation of medicine for treating and preventing lung/bronchial acute inflammation diseases

A trefoil factor, acute inflammation technology, applied in the field of biomedicine, can solve the problem of unknown disease protection effect, and achieve the effect of reducing respiratory tissue damage, reducing risk and damage, and inhibiting inflammatory response

Active Publication Date: 2016-05-18
SHANDONG RUIYING PIONEER PHARMA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, the protective effect of TFF2 on diseases caused by acute airway inflammation remains unknown

Method used

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  • Application of trefoil factor 2 in preparation of medicine for treating and preventing lung/bronchial acute inflammation diseases
  • Application of trefoil factor 2 in preparation of medicine for treating and preventing lung/bronchial acute inflammation diseases
  • Application of trefoil factor 2 in preparation of medicine for treating and preventing lung/bronchial acute inflammation diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1: Establishment of mouse models infected with highly pathogenic influenza and low pathogenic influenza

[0020] The present embodiment adopts high dose of influenza virus H7N9 strain A / Shanghai / 4664T / 2013 (3.5*10 5 Infection amount of half tissue cells TCID50 / 50ul volume) and H9N2 virus strain A / Chicken / Shanghai / F / 98 (1.7*10 7 EID50 / 50ul (volume of half of chicken embryos) was used to infect C57 mice by intranasal drops, and 40ul / mouse was anesthetized with ketamine before infection. Put the mice into IVC cages, weigh the mice for 14 consecutive days, and observe the survival and survival status of the mice. 6 hours, 1 day, 2 days, 3 days, 7 days, and 14 days after infection, lung tissues were taken, frozen in liquid nitrogen, and used for later use ( figure 1 A). figure 1 As shown in B, the body weight of H7N9-infected mice continued to decrease. On the 7th day, most of the mice lost more than 30% of their body weight, while the H9N2-infected mice lost 12.6...

Embodiment 2

[0021] Example 2: Transcriptome chip analysis and screening of significant differential gene expression in lung tissue of mice infected with severe influenza and mild influenza, and TFF2 was screened as a protective molecule

[0022] In this example, the Agilent2100 system was used to detect the genome-wide expression profiles of the lung tissues of mice infected with H7N9 and H9N2 at the time points of 0 day, 6 hours, 1 day, 2 days, 3 days, 7 days and 14 days. like figure 2 A, We set the P value as 0.01, and screened 188 genes with significant differences, among which 85 were highly expressed in the H7N9 group, and 103 were highly expressed in the H9N2 group. The highly expressed genes in the typical H7N9 group include: Olfr11, IFN13, IFNb1, Krt17, these genes are related to inflammation; while the highly expressed differential genes in the H9N2 group include TFF2, Gp2, Muc5b, Muc5a, Clca3, these genes are related to host defense and epithelial cells Restoration is closely ...

Embodiment 3

[0023] Example three: Cloning of TFF2 gene and eukaryotic expression of TFF2 protein

[0024] This embodiment first clones the gene according to the TFF2 sequence (the nucleotide sequence of human TFF2 protein (hTFF2) is shown in SEQ ID NO: 3, the amino acid sequence is shown in SEQ ID NO: 4, and the nucleotide sequence of mouse TFF2 protein (mTFF2) is shown in SEQ ID NO : 5, the amino acid sequence is shown in SEQIDNO: 6), after transfecting the recombinant pSV1.0-TFF2 or pSV1.0-TFF2-6xHis plasmid into 293T cells, the eukaryotic expression of the TFF2 protein was detected by WB, The nickel column was eluted with different concentration gradients of imidazole, the target protein was collected, filtered and washed to obtain high-purity TFF2 protein. The specific steps are as follows:

[0025] The cDNA generated by reverse transcription of RNA extracted from the lung tissue of mice infected with H9N2 was used as a template for PCR amplification. The upstream primer was SEQ ID NO...

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Abstract

The invention belongs to the technical field of biomedicine and relates to an application of TFF2 protein in preparation of a medicine for treating and preventing lung / bronchial acute inflammation diseases. In the invention, the significantly different gene expression of the lung tissues of mice infected by severe flu and mild flu are analyzed and screened by a transcriptome chip, and the screened TFF2 is a protective factor; and according to in-vitro synthesis of TFF2 protein, the survival rate of the flu-infected mice can be remarkably increased by dropwise adding the TFF2 protein while the weight loss of mice is reduced. Further study shows that the TFF2 does not influence the flu virus replication but can reduce the secretion of inflammatory factors, promote the repair of the pulmonary mucosa and reduce the damage of lung tissues. Since the mechanism of other respiratory pathogens or physical and chemical factors causing respiratory tissue damage is similar to that of flu virus infection and the protection of TFF2 on flu virus infection is not specific protection but broad-spectrum protection, the TFF2 plays an important protection role in an acute inflammation damage model induced by respiratory pathogens or other factors.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and specifically relates to the application of trefoil factor 2 (TFF2) in the preparation of medicines for the treatment and prevention of lung / bronchial acute inflammatory diseases caused by respiratory pathogen infection or other factors. Background technique [0002] Acute inflammatory diseases of the respiratory tract can be seen in respiratory tract infection, inhalation of high temperature, low temperature or toxic gases, etc. Among them, acute inflammatory diseases caused by respiratory tract infection are the most common, and viral infections are the most common. [0003] Respiratory tract infection pathogens have the characteristics of many types of pathogens, the general susceptibility of the population, fast transmission speed, strong infectivity and difficult control. Once an outbreak occurs, it will seriously threaten human health. In recent years, the epidemic caused by acute re...

Claims

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Application Information

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IPC IPC(8): A61K38/18A61P11/00A61P29/00
CPCA61K38/18
Inventor 徐建青张晓燕傅卫辉
Owner SHANDONG RUIYING PIONEER PHARMA
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