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Medical application of miR-574-5p to diabetes and related diseases thereof

A technology for diabetes and diseases, applied in the field of biomedicine, can solve problems such as unclear effects and mechanisms

Active Publication Date: 2020-11-10
GENERAL HOSPITAL OF THE NORTHERN WAR ZONE OF THE CHINESE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, its role and mechanism in the development of obesity and diabetes are still unclear

Method used

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  • Medical application of miR-574-5p to diabetes and related diseases thereof
  • Medical application of miR-574-5p to diabetes and related diseases thereof
  • Medical application of miR-574-5p to diabetes and related diseases thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0057] Example 1 The expression of miR-574-5p is reduced in the serum of obese people with insulin resistance and mice.

[0058] (1) Extraction of total RNA from each mouse tissue.

[0059] Normal C57BL / 6 mice were sacrificed by neck dislocation under isoflurane anesthesia, 100 mg of each tissue was extracted, and total RNA was extracted using TRIzol reagent (Invitrogen, Carlsbad, CA). Using PrimeScript with gDNA Eraser TM RT kit (TakaRa) and specific RT primers (Guangzhou Ruibo Biological Co., Ltd.), reverse transcribed 2 μg of total RNA into cDNA. In CFX96Touch TM On the real-time PCR detection system (Hercules), use Fast qPCR mixture (TakaRa) and specific primers (Guangzhou Ribo Biotech Co., Ltd.) were used for real-time quantitative PCR. U6 small nuclear RNA was used as an internal standard for miRNA. Each reaction was repeated three times and analyzed by the 2-ΔΔCt method.

[0060] (2) It was found that miR-574-5p had specific high expression in adipose tissue of m...

Embodiment 2

[0065] Example 2 Fat damage causes miR-574-5p expression to decrease.

[0066] (1) Differentiated 3T3L1 cells were treated with palmitic acid (PA).

[0067] After PA (400 mM) was added to the supernatant of 3T3L1 cells differentiated in vitro for 7 days and cultured for 24 hours, the cells were collected, RNA was extracted, and the expression of miR-574-5p was detected. It was found that PA treatment significantly reduced miR-574-5p expression, as figure 2 As shown in A.

[0068] (2) Obesity model of C57 mice treated with HFD.

[0069] Male C57BL / 6 mice (8-10 weeks old) were purchased from Nanjing Model Animal Center (Nanjing, China). This study was approved by the Northern Theater Command General Hospital Institutional Animal Care and Use Committee. All animal experimental protocols were in accordance with the Guide for the Care and Use of Laboratory Animals published by the National Institutes of Health. Mice were fed a high-fat diet (HFD) for 12 weeks and then divided...

Embodiment 3

[0072] Example 3 miR-574-5p initiates the differentiation of preadipocytes into adipocytes.

[0073] To clarify whether miR-574-5p is involved in adipocyte differentiation. Transfect overexpression of miR-574-5p vector (miR-574-5p mimic) or miR-574-5p inhibitor (inhibitor) in differentiated 3T3L1 cells, and detect its effect on 3T3L1 cell differentiation.

[0074] (1) Cell transfection.

[0075]X-tremeGENE HP DNA transfection reagent (invitrogen) was used to transfect miRNA mimic (100nM), miRNA inhibitor (100nM) and its control (Guangzhou Ruibo Biotech Co., Ltd.) into 3T3-L1 cells. After culturing for 48 hours, the cells were collected, proteins were extracted, and Western blot was performed to detect adipocyte differentiation marker proteins.

[0076] (2) Oil red staining of cells.

[0077] Oil red O solution was purchased from Sigma. Mix 4ml of Oil Red O solution with 6ml of deionized water to get 10ml of Oil Red O working solution. The working solution of Oil Red O mus...

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Abstract

The invention belongs to the field of biomedicine, and relates to medical application of miR-574-5p, in particular to application of the miR-574-5p to preparation of drugs for preventing or treating diabetes and related diseases thereof. The invention provides application of the miR-574-5p and a recombinant vector or a recombinant cell for expressing miR-574-5p nucleic acid molecules to preparation of the drugs for preventing or treating the diabetes and the related diseases thereof. The invention also provides application of a reagent for detecting the miR-574-5p or an active expression levelof the miR-574-5p in serum or plasma to preparation of a kit for predicting and / or evaluating the treatment effect and prognosis of the diabetes and the related diseases thereof. Experiments prove that the miR-574-5p is highly expressed in human adipose tissues, and the expression quantity is remarkably increased along with adipose cell differentiation; the miR-574-5p is increased to promote adipose cell differentiation and resist insulin resistance caused by obesity; the phenomena of the obesity and the insulin resistance can be aggravated by inhibiting the miR-574-5p expression; and in people with similar BMI, the miR-574-5p expression in serum of people with abnormal glucose tolerance and the diabetes can be obviously reduced. The miR-574-5p can be used for detecting, preventing or treating the diabetes and the related diseases.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to the medical use of miR-574-5p, in particular to the use of miR-574-5p in the preparation of drugs for the prevention or treatment of diabetes and related diseases. Background technique [0002] Diabetes mellitus is a metabolic disease characterized by chronic elevated blood sugar, mainly divided into type 1 and type 2, of which type 2 diabetes is particularly common. Type 2 diabetes is mainly characterized by insulin resistance with or without β-cell dysfunction. Diabetes is widely prevalent in the world. The World Health Organization reports that by 2035, the number of people with diabetes worldwide will exceed 600 million, and the global incidence of type 2 diabetes is 5.4%. The Diabetes Epidemiological Survey of the Diabetes Society of the Chinese Medical Association shows that in 2013, there were more than 139 million type 2 diabetes patients in my country, ranking first in the world...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61K48/00A61K45/00A61P3/10A61P5/50A61P1/16C12Q1/6883
CPCA61K31/7105A61K45/00A61P3/10A61P5/50A61P1/16C12Q1/6883C12Q2600/178C12Q2600/118C12Q2600/158C12Q2600/136
Inventor 韩雅玲闫承慧田孝祥李玉莹刘丹
Owner GENERAL HOSPITAL OF THE NORTHERN WAR ZONE OF THE CHINESE PEOPLES LIBERATION ARMY
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