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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 screened out as therapeutic targets, and corresponding therapeutic drugs can be developed, it will have a great impact on silicosis in my country. The status quo of disease treatment will be a powerful impetus

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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

Embodiment 1

[0036] Example 1: Reconstruction of mouse silicosis model

[0037] Establishment of mouse lung fibrosis animal model induced by silica dust: Select male C57BL / 6 mice with a body weight of 20-25g for 6-8 weeks, and perfuse 50mg / mL SiO2 prepared by 50μL normal saline through the bronchus under anesthesia. 2 The dust suspension, the control group was given normal saline, the mice were treated on the 7th, 14th, and 28th day, and the lung tissue was preserved. The right lower lung lobe was fixed in formaldehyde, embedded in paraffin, and sectioned for HE staining. The remaining lung tissues were quickly frozen in liquid nitrogen and stored in a -80°C refrigerator for later use. The protein levels of epithelial cell markers (E-cadherin) and mesenchymal cell markers (α-SMA, Vimentin) in lung tissue were determined by western blot. The results of mouse lung tissue pathological sections and fibrosis indicators were combined to evaluate whether the model of silica dust-induced lung fi...

Embodiment 2

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

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

[0049] (1) Preparation of RNA samples

[0050]Place the sterilized tissue homogenizer on ice, add 100 mg of mouse lung tissue, and then add 1.0 mL Trizol (Life Technologies / ambion, Carlsbad, CA) to grind until homogenized; ② Transfer the homogenate to 1.5 mL Add 200 μL of chloroform (trichloromethane) to the RNase EP tube, sandwich it between two plates and gently i...

Embodiment 3

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

[0056] (1) Animal model establishment

[0057] A chemically modified miRNA-489 agonist (agomir) was used to upregulate miRNA-489 levels in mice. Select male C57BL / 6 mice with a body weight of 20-25g for 6-8 weeks, and 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 perfusing 50μL normal saline through the bronchus under anesthesia 2 Dust suspension, in which the miRNA-489 high expression group was perfused with SiO 2 After the dust suspension, 5 nmol miRNA-489 agomir was injected into the trachea for the first time, and then 2.5 nmol miRNA-489 agomir was injected through the tail vein on 7, 14 and 21 days respectively, and lung tissue and serum were c...

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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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