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Application of microRNA-26a in preparation of drug for prevention or treatment of pulmonary fibrosis

1. Microrna-26a, pulmonary fibrosis technology, applied in gene therapy, drug combination, pharmaceutical formulation, etc., can solve the problem of unidentified key microRNA subtypes

Active Publication Date: 2013-11-20
HARBIN MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although more and more microRNAs have been discovered as biomarkers, determinants and therapeutic targets of human diseases, the key microRNA subtypes in pulmonary fibrosis have not been identified, their functions and disease-specific targets. The tropic route of drug delivery remains a challenge for researchers in the field

Method used

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  • Application of microRNA-26a in preparation of drug for prevention or treatment of pulmonary fibrosis
  • Application of microRNA-26a in preparation of drug for prevention or treatment of pulmonary fibrosis
  • Application of microRNA-26a in preparation of drug for prevention or treatment of pulmonary fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1, the expression of microRNA-26a is reduced in pulmonary fibrosis

[0021] In this experiment, tracheal injection of bleomycin was used to establish a model of pulmonary fibrosis in C57BL / 6 mice. Choose healthy adult C57BL / 6 mice aged 2-3 months, anesthetized with 1% pentobarbital sodium, expose the trachea, and inject bleomycin (BLM) or saline (Saline) into the trachea, 5mg / kg body weight or 50ul / 30g body weight, the pulmonary fibrosis model was formed after 28 days.

[0022] The lung tissue of the mice was fixed in 4% paraformaldehyde solution for pathological staining. It was found that the mice injected with BLM had pulmonary fibrosis, while the mice injected with normal saline had no fibrosis. The determination of hydroxyproline content found that compared with the saline group, the hydroxyproline content in the lung tissue of the mice in the BLM group was significantly increased. RNA was routinely extracted from mouse lung tissue, and real-time PCR rev...

Embodiment 2

[0024] Example 2. Inhibiting the expression of microRNA-26a can induce the occurrence of pulmonary fibrosis

[0025] Healthy adult C57BL / 6 mice aged 2-3 months were selected, and cholesterol-modified antagomiR-26a (cholesterol was linked to the 5' end of antagomiR-26a to increase its transmembrane property and specifically inhibit the expression of miR-26a) was tested. Tracheal injection, divided into three groups (1) injection of normal saline group (saline group); (2) injection of cholesterol-modified antagomiR-26a group; (3) injection of cholesterol-modified antagomiR-NC group (negative control). The injection dose was 10 mg / kg, the time point was the 1st, 2nd, 8th, 9th, 15th, and 16th day, and the samples were taken three weeks later. It was found that after the injection of cholesterol-modified antagomiR-26a, the collagen deposition in the lung tissue of the mice increased significantly, and the hydroxyl Mice with elevated proline levels developed lung fibrosis (*p<0.05 v...

Embodiment 3

[0027] Example 3. Expression of miR-26a can significantly inhibit pulmonary fibrosis in mice

[0028] Choose healthy adult C57BL / 6 mice aged 2-3 months, use cholesterol-modified microRNA-26a (agomiR-26a, connect cholesterol to the 5' end of microRNA-26a complementary link, increase the transmembrane property of microRNA, and overexpress miR -26a) transtracheal injection, divided into four groups (1) injection of normal saline group (saline group); (2) injection of BLM group; (3) injection of BLM+agomiR-26a group; (4) injection of BLM+agomiR-NC group (negative control). The injection dose of agomiR-26a / agomiR-NC is 10 mg / kg, and the injection time points are as follows: image 3 shown.

[0029] Inject cholesterol-modified agomiR-26a or agomiR-NC into the mouse trachea for 3 days, then inject BLM into the mouse trachea to create a mouse model of pulmonary fibrosis, and take samples after 4 weeks ( image 3 A-a), Masson staining results showed that early injection of agomiR-26...

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Abstract

The invention discloses the application of microRNA-26a in preparation of a drug for prevention or treatment of pulmonary fibrosis. According to the application of the microRNA-26a in preparation of the drug for prevention or treatment of the pulmonary fibrosis, experiments prove that microRNA-26a (miR-26a) expressions in experimental pulmonary fibrosis mice and clinical pulmonary fibrosis patients are substantially reduced; antagomiR-26a modified by cholesterol or LNA-AMO-26a modified by a locked nucleic acid (LNA) technology is used for specific inhibition of the miR-26a, and results show that mice pulmonary fibrosis can be caused; more importantly, the application further founds that: through use of the antagomiR-26a modified by the cholesterol or the LNA-miR-26a modified by the locked nucleic acid (LNA) technology for overexpression of the miR-26a in pulmonary tissues, the generation of the experimental pulmonary fibrosis can be effectively relieved; and the microRNA-26a modified by the cholesterol and the locked nucleic acid (LNA) are more safe relative to adenovirus transfection used in the experiments in the prior art. The application of the microRNA-26a in preparation of the drug for prevention or treatment of the pulmonary fibrosis provides a new pathophysiology mechanism for the generation of the pulmonary fibrosis, and provides the new drug for prevention and treatment of pulmonary fibrosis diseases.

Description

technical field [0001] The invention relates to a new medical application of endogenous non-coding small RNAs, in particular to the application of microRNA-26a in the prevention and treatment of pulmonary fibrosis, and belongs to the field of prevention and treatment of pulmonary fibrosis. Background technique [0002] Idiopathic pulmonary fibrosis (IPF) is a chronic interstitial lung disease of unknown etiology. Causes loss of lung elasticity and alveolar surface area, resulting in impairment of gas exchange and lung function. Its clinical manifestations were progressive dyspnea accompanied by irritating dry cough, and eventually died of respiratory failure. The incidence of IPF is high, the survival rate is low, and the prognosis is poor. After diagnosis, the average survival rate is 3 to 5 years, and its 5-year survival rate does not exceed 50%, which is extremely harmful to human health and life. However, the pathogenesis of pulmonary fibrosis remains unclear and there...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P11/00
Inventor 杨宝峰单宏丽梁海海吕延杰
Owner HARBIN MEDICAL UNIVERSITY
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