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Application of miR-127 in preparation of medicines for treating muscle diseases

A mir-127, muscle disease technology, applied in the field of muscle disease treatment, can solve the problems of skeletal muscle stem cell survival rate, self-renewal ability and migration ability, inability to express dystrophin, etc.

Active Publication Date: 2016-06-22
THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Such as transplantation of autologous skeletal muscle stem cells into mdx mice, but the autologous transplanted skeletal muscle stem cells still cannot express dystrophin
If allografted skeletal muscle stem cells are used to compensate for the missing dystrophin protein in mdx mice, there will be the following problems: 1) host immune rejection; 2) survival rate, self-renewal ability and migration of skeletal muscle stem cells after transplantation Reduced ability

Method used

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  • Application of miR-127 in preparation of medicines for treating muscle diseases
  • Application of miR-127 in preparation of medicines for treating muscle diseases
  • Application of miR-127 in preparation of medicines for treating muscle diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: miR-127 is highly enriched in skeletal muscle tissue and its expression level increases with the differentiation of C2C12 cells and skeletal muscle stem cells.

[0054] 1. Northern blotting was used to detect the expression abundance of miR-127 in various tissues and organs of 8-week-old mice. 8-week-old mice were selected, and lung, liver, heart, spleen, skeletal muscle, kidney, pancreas, small intestine, brain, and stomach tissues were collected respectively. RNA was first extracted separately, and then Northern Blot hybridization was performed. Specific steps are as follows:

[0055] (1) RNA extraction

[0056] Take 50-100mg of fresh or frozen tissue material, add liquid nitrogen in a mortar and grind it into powder, transfer the powder to a 15mL centrifuge tube with a small amount of liquid nitrogen, add 5mL Trizol to homogenate, let stand at room temperature for 5min; add 1mL chloroform (per mL Trizol (plus 0.2 mL chloroform) was shaken and mixed, and...

Embodiment 2

[0085] Example 2: miR-127 promotes the differentiation of C2C12 cells.

[0086] The expression profile of miR-127 in Example 1 suggests that it may play an important role in muscle cell differentiation. Therefore, a stable transfected cell line with high expression of miR-127 (about 12 times) was firstly constructed, and the cell line was used for functional detection.

[0087] Take 2×10 4 / cm2 The density of C2C12 cells stably expressing miR-127 was planted, and the differentiation medium was changed after 24 hours of culture. After 24 hours and 36 hours of culture, the early differentiation marker molecule myogenin (Myogenin, MyoG) and the late differentiation marker molecule myosin heavy chain ( MHC) expression. Cell immunofluorescence staining found that after 24 hours of differentiation, the overexpression of miR-127 significantly increased the number of MyoG positive cells, and the expression of MyoG significantly increased at the transcriptional and protein levels; af...

Embodiment 3

[0108] Example 3: miR-127 promotes the process of skeletal muscle injury regeneration.

[0109] (1) Establishment of skeletal muscle injury regeneration model

[0110] miR-127 transgenic mice were constructed from Nanjing model animals. In order to determine the function of miR-127 in bone injury regeneration, a CTX injury regeneration model of miR-127 mice was first constructed; mice were injected with 50 μL×10 μMCTX in the tibialis anterior muscle of the right leg, and the same amount of PBS was injected into the left leg as a control.

[0111] After skeletal muscle injury, there is an inflammatory response first, a large number of inflammatory factors are released, and the skeletal muscle stem cells in a resting state are activated. The activated skeletal muscle stem cells proliferate in large numbers and then enter the differentiation program to fuse with the damaged muscle fibers. This model uses the number of MyoD-positive cells to evaluate the number of activated skele...

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Abstract

The invention discloses application of miR-127 in preparation of medicines for treating muscle diseases. The invention provides application of any one of substances (1), (2) and (3) in preparation of medicines for treating muscle diseases: (1) miR-127; (2) a recombinant vector containing a coding gene of miR-127; and (3) a recombinant virus containing the coding gene of miR-127. The miR-127 can be used for preparing medicines for treating muscle diseases so as to effectively treat the muscle diseases.

Description

technical field [0001] The invention relates to the field of treatment of muscle diseases, in particular to the use of miR-127 in the preparation of medicines for treating muscle diseases. Background technique [0002] The life activities of bone organisms are inseparable from the movement and supporting functions of skeletal muscles. Not only that, the substances such as sugar and lipid ingested by our body are metabolized in skeletal muscle to produce energy, which plays a very important role in maintaining the stable state of the whole body. Patients with many diseases, such as tumors, HIV infection, and chronic heart failure, are often accompanied by symptoms of skeletal muscle loss in the late stage. [0003] A remarkable feature of skeletal muscle tissue is the ability to regenerate and repair after injury. This process is highly coordinated, and skeletal muscle stem cells (satellite cells) play an important role in this process. Skeletal muscle stem cells are locate...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61K31/7105C12N5/10A61P21/00
CPCA61K31/7105A61K48/00
Inventor 朱大海张勇翟丽丽武日茂
Owner THE INST OF BASIC MEDICAL SCI OF CHINESE ACAD OF MEDICAL SCI
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