Application of miRNA-489 to preparation of medicines for treating silicosis

A technology of mirna-489, 1. mirna-489 is applied in the application field of miRNA-489 in the preparation of silicosis drugs, and achieves the effects of quantitative accuracy, improving affinity and increasing stability

Inactive Publication Date: 2016-08-24
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, different from idiopathic pulmonary fibrosis, there are no reports of specific miRNAs for the treatment of silicosis. If tissue miRNAs specific or abnormally expressed in silicosis can be screen

Method used

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  • Application of miRNA-489 to preparation of medicines for treating silicosis
  • Application of miRNA-489 to preparation of medicines for treating silicosis
  • Application of miRNA-489 to preparation of medicines for treating silicosis

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0036] Example 1: Reconstruction of mouse silicosis model

[0037] Silica dust-induced mouse lung fiber animal model: select male C57BL / 6 mice weighing 20-25g for 6-8 weeks, and infuse 50μL of normal saline with 50mg / mL SiO under anesthesia. 2 For the dust suspension, the control group was given normal saline. The mice were treated on the 7, 14 and 28 days, and the lung tissue was retained. The right lower lung lobe was fixed with formaldehyde and embedded in paraffin and sectioned for HE staining. The remaining lung tissues were quickly frozen in liquid nitrogen and stored in a refrigerator at -80°C for later use. Western blot method was used to determine the protein levels of epithelial cell markers (E-cadherin) and mesenchymal cell markers (α-SMA, Vimentin) in lung tissue. The results of pathological sections of mouse lung tissues and fibrosis indicators are combined to evaluate whether the establishment of a mouse lung fibrosis model caused by silica dust is successful.

[00...

Example Embodiment

[0047] Example 2: Reconstruct the qRT-PCR experiment of miRNA-489 levels in a mouse silicosis model

[0048] After confirming that the model was established successfully, RNA was extracted from the lung tissues of 5 mice in the silica dust treatment group on the 7th, 14th, and 28th day and the normal saline group on the 28th day according to the lung tissue pathological section for qRT-PCR detection. Strict quality control was implemented throughout the research process, and each sample was tested three times in a row. All samples were blinded, that is, they were completed without the background of the samples to avoid bias.

[0049] (1) Prepare RNA samples

[0050] Place the sterilized tissue homogenizer on ice, put in 100mg mouse lung tissue, and then add 1.0mL Trizol (LifeTechnologies / ambion, Carlsbad, CA) to grind to a homogenized state; ②Transfer the homogenate to 1.5mL without Add 200μL of chloroform (trichloromethane) to the RNase EP tube, sandwich it between the two plates a...

Example Embodiment

[0055] Example 3: Animal experiment of pulmonary fibrosis in mice caused by silica dust that up-regulates the level of miRNA-489

[0056] (1) Animal model establishment

[0057] A chemically modified miRNA-489 agonist (agomir) was used to up-regulate the level of miRNA-489 in mice. Choose 6-8 weeks of male C57BL / 6 mice weighing 20-25g, divide the mice into control group (normal saline), silica dust group (SiO 2 Suspension), miR-NC+silica dust group (miR-NC+SiO 2 Suspension), miRNA-489 high expression + silica dust group (miRNA-489agomir+SiO 2 Suspension), 50mg / mL SiO prepared by bronchial infusion of 50μL of normal saline under anesthesia 2 Dust suspension, in which the high expression group of miRNA-489 is perfused with SiO in the bronchus 2 After the dust suspension, 5nmol miRNA-489 agomir was injected into the trachea for the first time, and then 2.5nmol miRNA-489 agomir was injected through the tail vein at 7, 14 and 21 days, respectively. Lung tissue and serum were collected on...

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Abstract

The invention provides application of miRNA-489 to preparation of medicines for treating silicosis. The sequence of miRNA-489 is 5'-AAUGACACCACAUAUAUGGCAGC-3'. Experiments prove that whether at the in-vitro cell level or at the in vivo mice silicosis model level, up-regulation of the expression of miRNA-489 has obvious inhibiting effects on silicious dust induced lung inflammations and fibrosis, which prompts that miRNA-489 can become a brand-new target for treating silicosis. miRNA-489 is conducive to research and development of the medicines for treating silicosis and provides a reference for development of medicines for treating other diseases.

Description

technical field [0001] The invention belongs to the field of genetic engineering and life sciences, in particular to the field of gene therapy, and relates to the application of miRNA-489 in the preparation of medicines for treating silicosis. Background technique [0002] Silicosis is a progressive, disabling, incurable fibrosis of lung tissue caused by the accumulation of free silica-containing dust in the lungs. At present, silicosis is still one of the most serious occupational diseases in my country. Data from the National Occupational Disease Reporting System shows that in 2014, a total of 29,972 occupational diseases were reported nationwide, and 26,873 new cases of pneumoconiosis accounted for 89.66% of the total number of occupational disease reports in 2014. There were 13,846 and 11,471 cases of coal workers' pneumoconiosis and silicosis, accounting for 94.21% of the total number of pneumoconiosis cases. Once silicosis occurs, even if there is no more exposure to s...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61P11/00
CPCA61K31/7105
Inventor 倪春辉吴秋云韩磊严玮文吉晓明刘易
Owner NANJING MEDICAL UNIV
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