Application of reagent for down-regulating Czib gene expression in preparation of medicine for treating or improving pulmonary fibrosis

A gene expression and pulmonary fibrosis technology, applied in the field of biomedicine, can solve problems such as obvious side effects and achieve the effect of inhibiting the occurrence of pulmonary fibrosis

Pending Publication Date: 2022-03-01
HENAN NORMAL UNIV
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  • Abstract
  • Description
  • Claims
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AI Technical Summary

Problems solved by technology

Two anti-fibrotic drugs approved by the US FDA, Nintedanib (nintedanib: a small molecule tyrosine kinase inhibitor) and Pirfenidone (pirfenidone: a growth factor and collagen precursor production inhibitor) are only effective in slowing down the disease. Exacerbation has some positive effects, but both drugs have significant side effects

Method used

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  • Application of reagent for down-regulating Czib gene expression in preparation of medicine for treating or improving pulmonary fibrosis
  • Application of reagent for down-regulating Czib gene expression in preparation of medicine for treating or improving pulmonary fibrosis
  • Application of reagent for down-regulating Czib gene expression in preparation of medicine for treating or improving pulmonary fibrosis

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Experimental program
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Effect test

Embodiment 1

[0059] Expression changes of Czib in a bleomycin-induced mouse model of lung fibrosis.

[0060] The construction of the bleomycin mouse model: the mice were anesthetized by intramuscular injection of pentobarbital sodium (30 mg / kg). Draw out 0.3ml of bleomycin (Bleo) normal saline solution (dissolved in 0.2-0.3 normal saline according to the ratio of 5 mg / kg) with the syringe, align the needle tip of the syringe with the cricoid cartilage space, penetrate the trachea and draw back the air, then raise the animal with the left hand With the head at a certain angle, push the needle with the right hand to inject slowly along the back wall of the trachea, and then continue to inject 0.3ml of air, then immediately erect the animal, rotate left and right, and pat the mouse on the chest and back to promote the uniform distribution of the drug in the lungs, the control group (Sal) Inject an equal volume of normal saline in the same way. After 21 days, the samples were taken, part of w...

Embodiment 2

[0062] Expression of Czib in bleomycin-induced A549 cells.

[0063] Construction of A549 cell injury model: A549 cells were treated with bleomycin for 72 hours, the total protein was extracted, and the expressions of Czib and fibrosis marker protein aSMA were detected by qPCR and Western blot. The result is as figure 2 The up-regulated expression of aSMA indicated that the induction of the A549 cell injury model was successful, and the expression of Czib was significantly up-regulated in this cell model.

Embodiment 3

[0065] Preparation of effective Czib siRNA.

[0066] Using siRNA design software, according to GenBank to obtain the Czib mRNA sequence of rats (NM_001304759.2), search the AA sequence and record the 19 nucleotides adjacent to the 3' end of each AA, and screen out the GC content between 30-55% siRNA. Further screening is carried out according to the siRNA design principles, and the homology of the screened siRNA sequences is searched by BLAST in the genome database of GenBank, and sequences with more than 3 base mismatches with non-homologous genes are selected to exclude For the possibility of non-specific inhibition, on the basis of meeting the above conditions, three siRNAs that inhibit the expression of Czib were screened out (see Table 1).

[0067] Table 1 siRNA sequences targeting Czib

[0068]

[0069]

[0070] Note: In the siRNA sequence, 5'-3' is the sense strand, and 3'-5' is the antisense strand.

[0071] Synthesize the siRNA sequences shown in Table 1, and...

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Abstract

The invention discloses application of a reagent for down-regulating Czib gene expression in preparation of a medicine for treating or improving pulmonary fibrosis, relates to the technical field of biomedicine, and provides that the gene Czib has a remarkable relationship with the occurrence of pulmonary fibrosis for the first time by researching a lung fiber regulation function gene. It is found that the effect of inhibiting pulmonary fibrosis can be achieved by down-regulating the expression level of the Czib gene. In addition, a specific target spot aiming at Czib is screened, the siRNA designed according to the target spot can effectively inhibit bleomycin-induced A549 cell injury, apoptosis, mitochondrial membrane potential drop and cell epithelial-mesenchymal transition, and a new way and research direction are provided for treatment of pulmonary fibrosis and cell injury and inhibition of cell epithelial-mesenchymal transition.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to the application of a reagent for down-regulating Czib gene expression in the preparation of medicines for treating or improving pulmonary fibrosis. Background technique [0002] Pulmonary fibrosis, the terminal outcome of a variety of interstitial lung diseases, is a chronic, progressive, lethal disease characterized pathologically by destruction of the lung parenchyma, marked changes in alveolar epithelial cell phenotype, deposition of extracellular matrix, and fibroblast Abnormal proliferation and accumulation. More than 200 reasons are now known to cause pulmonary fibrosis, such as genetic diseases, autoimmune diseases, exposure to toxic substances in the environment, drugs and radiation, etc. Among them, pulmonary fibrosis without a cause is called idiopathic pulmonary fibrosis Idiopathic Pulmonary Fibrosis (IPF). According to statistics, there are about 5 million new I...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/713A61P11/00
CPCA61K31/713A61P11/00
Inventor 张春艳赵亚彬余梦丽刘萍段若雨
Owner HENAN NORMAL UNIV
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