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Application of long-chain non-coding RNA MRAK053938 in the preparation of drugs for treating pulmonary fibrosis

A long-chain non-coding, pulmonary fibrosis technology, applied in the direction of DNA / RNA fragments, gene therapy, drug combination, etc., can solve the problems of ineffective inhibition of pulmonary fibrosis and aggravation of pulmonary fibrosis

Inactive Publication Date: 2015-09-23
BINZHOU MEDICAL COLLEGE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Traditional treatment is mainly based on corticosteroids. Clinical research results show that these drugs cannot effectively inhibit the occurrence of pulmonary fibrosis, but can aggravate the degree of pulmonary fibrosis in some severe patients. It is urgent to find new and effective treatments for pulmonary fibrosis Methods

Method used

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  • Application of long-chain non-coding RNA MRAK053938 in the preparation of drugs for treating pulmonary fibrosis
  • Application of long-chain non-coding RNA MRAK053938 in the preparation of drugs for treating pulmonary fibrosis
  • Application of long-chain non-coding RNA MRAK053938 in the preparation of drugs for treating pulmonary fibrosis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Expression of lncRNA-MRAK053938 in normal lung tissue and fibrotic tissue

[0022] 1. Materials SD rats, male, weighing 200 ± 20 grams, clean grade, provided by the Experimental Animal Center of Shandong Luye Pharmaceutical Co., Ltd., certificate number: SYXK (Lu) 20030020, in a clean grade animal observation room, adaptive feeding 3 days; normal lung epithelial cell line (RLE-6TN), fibroblast cell line (RL1) were purchased from ATCC cell bank; RPMI1640 medium, high-glucose DMEM medium (DMEM-LG), fetal bovine serum (fetal bovine serum, FBS) were purchased from Hyclone Company of the United States; TGF-β1 was purchased from Invitrogen Company; riboFECT TM CP transfection reagent (Ruibo Biotechnology Co., Ltd., Guangzhou, China); SYBR Green PCR Master Mix (TAKARA, Dalian, China).

[0023] 2. Method

[0024] 1. Prepare rat pulmonary fibrosis model: 30 SD rats are divided into normal group and model group at random, and rats are anesthetized with 4% chloral hy...

Embodiment 2

[0042] Example 2. Effects of siRNA interference fragments specific to MRAK053938 on transdifferentiation of myofibroblasts

[0043] 1. Cell culture and grouping: select RLE-6TN and RL1 cells in logarithmic growth phase, make single cell suspension with 0.25% trypsin, and inoculate in 6-well plate. A blank control group, a TGF-β1 stimulation group, a transfection group, and a transfection control group were set up; when the cells grew to 70-80% confluent, the transfection group and the transfection control group were treated with riboFECT respectively. TM CP transfection reagent transfected cells with siRNA interference fragment and control siRNA at a final concentration of 50nM. The blank control group was cultured with serum-free medium, and the other groups were stimulated with 5nM TGF-β1. Cells were collected after 72 hours of culture.

[0044] 2. Western blot detection of expression changes of α-SMA

[0045] Strictly follow the instructions of the Western and IP cell lys...

Embodiment 3

[0047] Compared with the blank control group, the expression of α-SMA in the TGF-β1 stimulation group was significantly up-regulated, indicating that TGF-β1 can induce the transformation of cells into myofibroblasts; compared with the TGF-β1 stimulation group, RLE-6TN / siRNA-MRAK053938 The expression of α-SMA in the group and RL1 / siRNA-MRAK053938 group was significantly down-regulated, with a difference of 3.43 and 1.79 times, respectively, suggesting that reducing the expression of lncRNA-MRAK053938 can inhibit the transdifferentiation of RLE-6TN cells and the activation of RL1 cells (see figure 2 ). Example 3. Effects of siRNA interference fragments specific to MRAK053938 on the migration of myofibroblasts

[0048] 1. Cell scratch test: select cells RLE-6TN and RL1 in the logarithmic growth phase, make a single-cell suspension with 0.25% trypsin, and inoculate them on a special culture dish of the living cell workstation. Set up TGF-β1 stimulation group, transfection contro...

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Abstract

The invention relates to an application of a long-chain non-coding RNA-MRAK053938 gene sequence in prevention and treatment of idiopathic pulmonary fibrosis. An interference sequence, which is specifically targeted on lnc RNA-MRAK053938, is designed and then is transfected to cells to reduce the expression of lnc RNA-MRAK053938 so as to prevent and treat pulmonary fibrosis.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and specifically relates to the application of a long-chain non-coding RNA-MRAK053938 gene sequence in the prevention and treatment of idiopathic pulmonary fibrosis. Background technique [0002] Long noncoding RNA (long noncoding RNA, lncRNA) is a class of RNA molecules with a length of more than 200nt discovered for the first time when Okazaki et al. sequenced the full-length complementary DNA (cDNA) library of mice in 2002, accounting for 4 % to 9%, they lack protein coding function, are located in the nucleus or cytoplasm, and have obvious cell, tissue specificity and developmental stage specificity. As a new class of non-coding RNA with regulatory functions, lncRNAs have attracted extensive attention from researchers for their powerful regulatory functions and complex regulatory mechanisms. With the continuous discovery of lncRNAs, researchers are paying more and more attention to their e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113A61K48/00A61P11/00
Inventor 吕长俊宋晓冬张瑾锦曲桂武
Owner BINZHOU MEDICAL COLLEGE