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Application of mir‑127 inhibitors in anti-inflammatory and lung injury protection drugs

A 1. mir-127, anti-mir-127 technology, applied in the field of biomedicine, can solve the problem of unclear and imperfect research on the immune response regulation mechanism, so as to reduce the production of pro-inflammatory factors and weaken the inflammatory response. , The effect of preventing lung inflammation and damage

Active Publication Date: 2017-11-14
HANGZHOU NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many miRNAs have been found to be related to the immune response, many researches on the regulatory mechanism of miRNA on the immune response are not clear and not perfect, and there is no report on the regulatory role of miR-127 in the inflammatory response.

Method used

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  • Application of mir‑127 inhibitors in anti-inflammatory and lung injury protection drugs
  • Application of mir‑127 inhibitors in anti-inflammatory and lung injury protection drugs
  • Application of mir‑127 inhibitors in anti-inflammatory and lung injury protection drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1: miR-127 can be activated by the LPS-TLR4-NF-κB signaling pathway

[0034] (1) Expression of miR-127 stimulated by different TLR ligands

[0035] First, we stimulated primary mouse peritoneal macrophages with TLR4 ligand LPS (100ng / mL), collected cells at 0, 2, 4, 8, 16, and 24 hours after stimulation, extracted RNA and reverse transcribed , Quantitative polymerase chain reaction (polymerase chain reaction, PCR) method was used to detect the dynamic changes of miR-127 expression level in macrophages. The results indicated that the activation of TLR4 could activate the expression of miR-127. The expression of miR-127 first decreased after 2 hours of LPS stimulation, down to 10-20% of the base value; then increased rapidly, and the expression of miR-127 was the highest after 16 hours of stimulation, up to 7 times of the base value around; then began to decline ( figure 1 a).

[0036] The above results prove that miR-127 can be activated by the LPS-TLR4 signa...

Embodiment 2

[0040] Example 2: miR-127 regulates the expression of inflammatory factors and the polarity transformation of macrophages

[0041] (1) The effect of miR-127 on the production of IL-6, TNF-α, IL-1β and IL-10 in macrophages activated by LPS

[0042] We examined the mRNA and protein levels of TLR4-triggered macrophage production of inflammatory and effector cytokines. Mouse primary macrophages were transfected with miR-127mimic and anti-miR-127 for 24 hours, stimulated with LPS (100ng / mL), collected cells at 0, 3, 6, and 12 hours after stimulation, extracted RNA and reacted Transcription, quantitative PCR method was used to detect the gene transcription levels of IL-6 and TNF-α. The results showed that after TLR4 activation in the control group, the gene transcription levels of IL-6 and TNF-α in macrophages reached the highest peak at 6 hours. 6 gene transcription levels ( image 3 a), the gene transcription levels of TNF-α and IL-1β also showed a certain upward trend ( imag...

Embodiment 3

[0052] Example 3: Bcl6 is the target gene of miR-127

[0053] (1) Expression of Bcl6 in LPS-stimulated macrophages

[0054] Through bioinformatics analysis, we found that there may be a specific target site paired with the miR-127 seed sequence region in the 3'UTR region of Bcl6. Subsequently, we stimulated mouse peritoneal macrophages with LPS (100ng / mL), collected cells at 0, 1, 2, 3, 4, and 12 hours after stimulation, prepared protein samples as described in the method section, and detected them by Western blot Changes in protein expression of Bcl6. The results showed that the expression of Bcl6 increased with the stimulation of LPS, and the expression was the highest at 2 hours, and then gradually weakened as the stimulation time increased. Compared with the previous experimental results, it shows that the protein change of LPS-induced Bcl6 and the expression of miR-127 just show the opposite expression law, which suggests that miR-127 may be involved in the expression r...

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Abstract

The present invention discloses the application of miR-127 inhibitor in the preparation of anti-inflammation and lung injury protection medicine. Inhibition of miR‑127 is effective in preventing lung inflammation and injury induced by lung injury. Inhibitors of miR-127 can reduce the production of pro-inflammatory factors, weaken the inflammatory response of the airway, and reduce the infiltration of inflammatory cells. The present invention discloses that miR-127 inhibitors can be used to prepare medicines for preventing acute lung injury and its complications, and the action mechanism of miR-127 inhibitors for preventing acute lung injury and its complications. The miR-127 inhibitor targets Bcl6 and inhibits LPS-induced inflammatory response by regulating the Bcl6 / Dusp1 / JNK pathway.

Description

technical field [0001] The invention belongs to the technical field of biomedicine and relates to the application of miR-127 inhibitors in the preparation of anti-inflammation and lung injury protection drugs. Background technique [0002] Acute lung injury is characterized by the release of a large number of inflammatory factors in the pulmonary interstitium, the accumulation of inflammatory cells, cellulose deposition and pulmonary edema. A dysregulated inflammatory response causes severe tissue damage and a continued deterioration of lung function. The causes of acute lung injury and its complications have not been thoroughly studied by the scientific community, nor have effective treatments been developed. Therefore, the mortality rate of acute lung injury is as high as 40%. [0003] microRNAs (miRNAs, small RNAs) are a class of single-stranded small RNA molecules with a length of 18-25 nucleotides, which mainly form RNA-induced silencing complexes (RNA-induced silenci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00A61P29/00A61P11/00
Inventor 史丽云应航洁卢哲
Owner HANGZHOU NORMAL UNIVERSITY
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