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Application of short peptide MPM in the preparation of medicine for treating diseases related to muscle cell differentiation

A muscle cell, short peptide technology, applied in the field of biomedicine, to achieve the effect of improving differentiation ability and great application prospects

Active Publication Date: 2019-11-05
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the role of the short peptide MPM in muscle development and functional regulation, and its application in the diagnosis and treatment of diseases related to muscle cell differentiation have not been reported yet.

Method used

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  • Application of short peptide MPM in the preparation of medicine for treating diseases related to muscle cell differentiation
  • Application of short peptide MPM in the preparation of medicine for treating diseases related to muscle cell differentiation
  • Application of short peptide MPM in the preparation of medicine for treating diseases related to muscle cell differentiation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Example 1 Expression changes of MPM in the process of myogenic differentiation of mouse myoblast cell line C2C12 cells

[0057] 1. Inducing myogenic differentiation of cells

[0058] (1) Inoculation of mouse myoblast cell line C2C12: C2C12 cells were inoculated at an appropriate density in a 6-well plate, and cultured in high-glucose DMEM containing 10% fetal bovine serum.

[0059] (2) Induction of myogenic differentiation: when the cells grew to 90% density, the cell culture medium was replaced with high-glucose DMEM containing 2% horse serum to induce myogenic differentiation. Use RNA extraction reagent TRIzol and protein extraction solution 1×SDS cell lysate to collect differentiation 0 days (GM), 1 day (DM 1), 2 days (DM 2), 3 days (DM 3), 4 days (DM 4 ), 5 days (DM 5), 6 days (DM 6) of RNA and protein.

[0060] 2. Western blot

[0061] (1) Before electrophoresis: prepare Tricine-SDS-PAGE (Tricine-SDS-polyacrylamide gel);

[0062] (2) Electrophoresis: The protei...

Embodiment 2

[0077] Example 2 Expression changes of MPM in the development of mouse skeletal muscle

[0078] 1. RNA and protein extraction

[0079] Skeletal muscle tissues of mice at different developmental stages (1 day, 7 days, 2 weeks, 3 weeks) were cut, and the tissue was lysed with TRIzol and 1×SDS cell lysate, and RNA and protein were extracted;

[0080] 2. Western blotting, the method is the same as step 2 of Example 1;

[0081] 3. Real-time quantitative PCR (qPCR) detection, method is the same as embodiment 1 step 3;

[0082] 4. Result analysis

[0083] Such as figure 2 As shown, the RNA and protein levels of MPM in the skeletal muscle tissues of 1-day-old, 7-day-old, 2-week-old, and 3-week-old mice were detected, and it was found that the expression of MPM gradually increased with the development of mouse skeletal muscle , suggesting that MPM plays a key role in the development of mouse skeletal muscle.

Embodiment 3

[0084] Example 3 The expression changes of MPM in the process of mouse skeletal muscle regeneration

[0085] 1. Injecting cardiotoxin to induce skeletal muscle regeneration in mice

[0086] Take 5-week-old C57BL / 6J mice, inject 50 μL of 20 μM cardiotoxin into the left gastrocnemius muscle, and inject 50 μL of 20 μM cardiotoxin into the right side as a control, collect the gastrocnemius muscles of 1 day, 3 days, 5 days, and 7 days after injection, and use TRIzol and 1 ×SDS cell lysate to lyse tissue, extract RNA and protein;

[0087] 2. Western blotting, the method is the same as step 2 of Example 1;

[0088] 3, real-time quantitative PCR (qPCR) detection, with embodiment 1 step 3;

[0089] 4. Result analysis

[0090] Such as image 3 As shown, the RNA and protein levels of MPM in the mouse skeletal muscle tissue of the gastrocnemius muscle of 1 day, 3 days, 5 days, and 7 days after the injection of cardiotoxin were detected, and it was found that the expression of MPM incr...

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Abstract

The invention discloses application of a short peptide MPM in the preparation of a medicine for treating diseases related to muscle cell differentiation. An amino acid sequence of the short peptide MPM is shown in SEQ ID NO. 1 or SEQ ID NO. 2, a nucleotide sequence of LINC00116 encoding the short peptide MPM is shown in SEQ ID NO. 3 or SEQ ID NO. 4. The application of the short peptide MPM in thepreparation of the medicine for treating the diseases related to the muscle cell differentiation finds that the short peptide MPM plays a key role in cell myogenic differentiation, skeletal muscle development and regeneration, muscle fiber development and regeneration; high MPM expression promotes cell myogenic differentiation, while low expression inhibits the myogenic differentiation of cells, and the expression level of MPM directly affects the maximum grip and mouse exercise tolerance. Compared with normal people, the expression level of the MPM is decreased in the diseased muscles of patients with tibial muscular dystrophy (DMD), Duchenne muscular dystrophy (TMD), rhabdomyosarcoma of different pathological types and the muscles of the elderly; and it indicates that the short peptide MPM is associated with muscle cell differentiation, the differentiation of the muscle cells can be promoted by promoting the expression of the MMP, and the medicine for treating diseases related to themuscle cell differentiation is prepared.

Description

technical field [0001] The present invention relates to the technical field of biomedicine, and more specifically, relates to the application of the short peptide MPM in the preparation of drugs for the treatment of diseases related to muscle cell differentiation. Background technique [0002] Muscle is the largest organ in the human body, and muscle differentiation is very important for muscle development and regeneration. The process of muscle differentiation is a process regulated by multiple genes. Mutations in any link will lead to muscle differentiation disorders and diseases related to muscle cell differentiation, such as sarcopenia, muscle atrophy, and rhabdomyosarcoma. [0003] Sarcopenia, also known as skeletal sarcopenia, is common in the elderly and is mainly manifested as a decrease in skeletal muscle mass and weakened muscle strength. With the increase of age, sarcopenia will eventually affect the functions of various organs in the elderly, including the decli...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61P21/00A61P35/00
CPCA61K38/1709A61P21/00A61P35/00
Inventor 庄诗美朱颖林仪芳
Owner SUN YAT SEN UNIV
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