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Renal fibrosis miRNA marker

A kidney fibrosis, miRNA-576 technology, applied in the direction of drug combination, microbial measurement/testing, biochemical equipment and methods, etc.

Active Publication Date: 2017-10-10
昆明医科大学第六附属医院
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Dysregulation of miRNA expression in cells can lead to the occurrence of various diseases including renal fibrosis. The latest research shows that some miRNAs are abnormally expressed in the kidney and urine of patients with renal fibrosis, but which miRNA is related to the occurrence of renal fibrosis , development has not yet reached a consensus

Method used

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  • Renal fibrosis miRNA marker
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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] This example demonstrates that miR-576 is lowly expressed in urine samples from people with renal fibrosis.

[0049] 1. miRNA extraction

[0050] Using Tiangen's miRNA extraction kit, add an equal volume of lysate to every 200 μl of urine, shake and mix for 30 seconds. After standing at room temperature for 5 minutes, centrifuge at 12,000rpm for 10min, take the supernatant, add 200μl chloroform, and shake vigorously for 15 seconds. After standing at room temperature for 5min, centrifuge at 12,000rpm for 15min. Water phase, RNA is mainly in the water phase, transfer the water phase to a new tube, slowly add 1 / 3 volume of absolute ethanol to mix well, transfer to the adsorption column together, place at room temperature for 2min, centrifuge at 12,000rpm for 30 seconds , retain the effluent. Slowly add 2 / 3 volume of absolute ethanol to the volume of the effluent, mix well, and transfer them to the adsorption column together. After standing at room temperature for 2 minut...

Embodiment 2

[0058] This example demonstrates that miRNA-576 is underexpressed in renal fibrosis cell lines.

[0059] 1. EMT cell model

[0060] EMT refers to epithelial-mesenchymal transition and is part of the process of renal fibrosis. Using the renal tubular epithelial cell line HK-2 cells, add TGF-β to the HK-2 cell culture medium to a final concentration of 10ng / mL, and take pictures after culturing for 48 hours: HK-2 cells without adding TGF-β have a typical Epithelial cells have cobblestone-like morphological characteristics, and the intercellular connections are tight; HK-2 added with TGF-β shows a spindle-shaped shape, with obvious intercellular spaces, and the cells are relatively thin, similar to the shape of fibroblasts, as shown in figure 2 shown.

[0061] 2. For miRNA extraction and reverse transcription, refer to the experimental process in Example 1.

[0062] 3.Q-PCR detection, refer to the experimental process in Example 1

[0063] 4. Results

[0064] like image 3...

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Abstract

The invention discloses a miRNA marker which is miRNA-576. The miRNA-576 can be used for judging renal fibrosis progress and severity degree. Inspection proves that the miRNA-576 can effectively distinguish a renal fibrosis sample from a non-renal-fibrosis sample. The miRNA-576 can be used for preparing a drug for inhibiting renal fibrosis on the basis. A novel diagnosis method is provided for clinically diagnosing renal fibrosis at molecular level, and a novel drug target is provided for renal fibrosis gene therapy.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a renal fibrosis miRNA marker and its application, in particular to a renal fibrosis-related miRNA-576 marker and its application. Background technique [0002] miRNA is a 21-22nt non-coding RNA molecule that naturally exists in the body. It is a class of RNA that regulates the expression of target genes through post-transcriptional gene silencing. It is estimated that about 1 / 3 of genes in organisms are regulated by miRNAs. The complex of miRNA and RISC can bind to the complementary sequence in the 5'-UTR or 3'-UTR of the target gene mRNA through base pairing, inhibit protein translation, or trigger mRNA degradation, thereby negatively regulating the expression of the target gene. [0003] Renal fibrosis (including renal interstitial fibrosis and glomerulosclerosis) is the main pathological basis of the final stage of renal damage caused by various reasons. The proliferation and activa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68A61K45/00A61P13/12
CPCA61K45/00C12Q1/6883C12Q2600/158C12Q2600/178
Inventor 朱芸滕兆伟孙阳杨芳卢永新
Owner 昆明医科大学第六附属医院
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