Use of miRNA-199a as an agent to inhibit adipogenic differentiation of myoblasts

A technology of miRNA-199a and myoblasts, which is applied in the direction of DNA/RNA fragments, recombinant DNA technology, and the use of vectors to introduce foreign genetic materials, etc., to achieve simple methods, high specificity and sensitivity, and important clinical and scientific research application value Effect

Active Publication Date: 2018-06-29
CHONGQING ACAD OF ANIMAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, the relationship and application of miRNA-199a and the adipogenic differentiation of muscle cells have not yet been researched and patented

Method used

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  • Use of miRNA-199a as an agent to inhibit adipogenic differentiation of myoblasts
  • Use of miRNA-199a as an agent to inhibit adipogenic differentiation of myoblasts
  • Use of miRNA-199a as an agent to inhibit adipogenic differentiation of myoblasts

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1 Drugs Stimulate Adipogenic Differentiation of C2C12 Muscle Cell Line

[0041] By "cocktail adipogenic induction method", C2C12 cells can be gradually transdifferentiated into typical adipocytes, and lose the characteristics of muscle cells ( figure 1 ). These changes include that the cell shape becomes spindle-shaped and gradually rounded, a large number of lipid droplets appear in the cell, the mRNA and protein levels of lipogenic genes FASn, C / EBPα, PPARγ, and Fabp4 in the cell are significantly increased, and the myocyte differentiation factor The expression levels of Myog, MyoD, and MYF5 were down-regulated.

[0042] Adipogenic differentiation of the C2C12 muscle cell line was stimulated by drugs, and miRNA expression differences between adipogenically differentiated C2C12 cells and normal myogenically differentiated C2C12 cells were screened by gene chips, and several major differentially expressed miRNAs were screened out, and then analyzed by bioinform...

Embodiment 2

[0043] Example 2 Screening and verification of adipogenic differentiation-related miRNAs by gene chip technology and quantitative PCR assay

[0044] Materials: adipogenically differentiated C2C12 cells, normal myogenically differentiated C2C12 cells and undifferentiated C2C12 cells.

[0045] Methods: Differentially expressed miRNAs in adipogenically differentiated C2C12 cells and normal myogenically differentiated C2C12 cells were screened by gene chip, and quantitative PCR was used to verify the results.

[0046] High-throughput gene chip detection was used to analyze the expression changes of intracellular miRNAs under different differentiation pathways. The gene chip results showed that the expression levels of 60 miRNAs were significantly different among adipogenic transdifferentiated cells, myogenic differentiated cells and undifferentiated cells ( figure 2 , one-way ANOVA, P<0.05)

[0047] Quantitative PCR was used for verification. The results showed that 10 miRNAs w...

Embodiment 3

[0048] Example 3 Verification that miR-199a inhibits the transdifferentiation of myoblasts into adipocytes

[0049] Transfected cells with synthetic mimic and inhibitor, overexpressed and silenced miR-199a in cells, and found that overexpressing miR-199a significantly inhibited the adipogenic differentiation of C2C12 cells ( image 3 , including the reduction of intracellular lipid droplets and down-regulation of lipogenesis gene expression), but silencing miR-199a had no significant effect on cell differentiation.

[0050] It shows that miR-199a has a negative regulatory effect on the adipogenic differentiation of myocytes.

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Abstract

The invention belongs to the technical field of biology, and relates to application of an miRNA-199a in a reagent for inhibiting adipogenic differentiation of myoblasts. A kit used for inhibiting the myoblasts to be trans-differentiated into adipocytes comprises an miRNA-199a simulant and an miRNA-199a carrier acceptable in pharmacy. According to the application of the miRNA-199a in a reagent for inhibiting adipogenic differentiation of the myoblasts, an Fatpl gene serves as a target gene of the miRNA-199a, low expression of the Fatpl gene indicates that adipogenic differentiation of the myoblasts is successfully inhibited when the kit is used, the miRNA-199a and the kit can be applied to genetic marker assistant breeding and improvement of meat quality reagents, and the important scientific research and application value is avhieved.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to the application of miRNA-199a in a reagent for inhibiting the transdifferentiation of myoblasts into adipocytes. Background technique [0002] microRNA (miRNA) is a kind of small non-coding RNA (non-coding RNA) widely present in animals, plants, bacteria and viruses. It guides the silencing complex to degrade target mRNA or inhibit translation through sequence complementarity. One miRNA can regulate multiple or even hundreds of target genes, and a gene may be regulated by multiple miRNAs at the same time. Through specific / systematic regulation of target genes, miRNAs are widely involved in and regulate various transcriptional activations and signal transductions in cell proliferation, differentiation, metabolism, senescence and apoptosis. [0003] Muscle and adipose tissue are two extremely important organs in humans and animals. In particular, they are both regulated by hormones such...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/113C12N15/85
Inventor 齐仁立黄金秀王琪王敬
Owner CHONGQING ACAD OF ANIMAL SCI
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