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Application of miR-20a to aspects of promoting nerve regeneration and repairing nerve injury

A technology of mir-20a and nerve injury, applied in the field of biomedicine, to achieve the effect of promoting growth

Pending Publication Date: 2020-08-11
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are few reports on the role of miR-20a in peripheral nerve injury. The present invention aims to further explore the regulation of differentially expressed miR-20a on DRG neurons during peripheral nerve injury repair.

Method used

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  • Application of miR-20a to aspects of promoting nerve regeneration and repairing nerve injury
  • Application of miR-20a to aspects of promoting nerve regeneration and repairing nerve injury
  • Application of miR-20a to aspects of promoting nerve regeneration and repairing nerve injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Investigating the expression changes of miR-20a in DRG tissue after rat sciatic nerve injury

[0055] 1. Preparation of rat sciatic nerve injury model

[0056] Adult healthy male C57BL / 6J mice, weighing 20 g, were randomly divided into 6 groups, 6 rats in each group. After compound anesthesia, the skin of the left lower limb was incised to expose the sciatic nerve parallel to the midsection of the femur on the left side, which was pinched with toothless forceps for 30 seconds, and then the skin was sutured. After the operation, the animals were fed under suitable conditions to relieve their pain, with 12 hours of light every day, and free access to water and food. 0 h, 3 h, 9 h, 1 d, 4 d, and 7 d after sciatic nerve crushing in rats were obtained from L4-L6 DRG tissues on the crushing side.

[0057] 2. RNA extraction and qRT-PCR of DRG tissue after rat sciatic nerve injury

[0058] L4-6 DRG tissues of SD rats at different time points after 0 h, 3 h, 9 h, ...

Embodiment 2

[0069] Example 2 Overexpression of miR-20a in vitro promotes axonal growth of DRG neurons

[0070] 1. Extraction of primary DRG neurons

[0071] DRG neurons come from adult male rats. The dorsal root ganglia are removed and placed in dissection solution HA, digested with an appropriate amount of 3.3 mg / ml collagenase, at 37 °C for 90 min. Discard the collagenase, add an appropriate amount of 0.25% trypsin to digest, 37 ℃, about 20min.

[0072] Trypsinization was terminated with PBS containing 10% fetal bovine serum, and the supernatant was discarded after centrifugation. To remove glial cells, suspend the cells with 15% bovine serum albumin, centrifuge, and discard the supernatant. Cells were resuspended in neuronal medium, passed through a 200-mesh sieve, and seeded into poly-lysine-coated plate wells.

[0073] 2. Transient electrostaining of neuronal cells

[0074] Use Rat Neuron Nucleofector™ Solution (Lonza Company) to suspend the digested DRG neuronal cells, add 1 μ...

Embodiment 3

[0078] Example 3 Overexpression of miR-20a in vivo promotes axonal growth of DRG neurons

[0079] Select adult healthy male rats, after compound anesthesia, shave the back of the rats, disinfect with iodine, cut the skin, cut the muscles at the ilium, expose the spine at L5, inject 30 μl of control with a BD needle Or Cy3-labeled agomir, a chemical mimic of miR-20a, after which muscle and skin were sutured. Rats were raised in a suitable environment, with free access to food and water, and 12 h of light every day. After 48 h, the second injection was performed, which was the same as the first injection. After another 72 hours, the sciatic nerve was crushed (the model preparation method of the sciatic nerve crush was the same as that in Example 1), the rat hair around the left leg was shaved off with a razor, the skin was cut after povidone iodine disinfection, and the muscle was further cut , exposing the left sciatic nerve. Generally, the upper end of the bifurcation is ...

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Abstract

The invention discloses application of miR-20a to preparation of a medicine for promoting nerve regeneration and repairing nerve injury, and further discloses application of the miR-20a to aspects ofpromoting DRG neuron axon regeneration and repairing peripheral nerve injury. The application is characterized by comprising the following processes: S1, the expression change of the miR-20a in DRG tissue after sciatic nerve injury of rats is detected; S2, the miR-20a is overexpressed in vitro to promote growth of DRG neuron axons; and S3, the miR-20a is overexpressed in vivo to promote growth ofthe DRG neuron axons. The miR-20a can participate in repairing of peripheral nerve injury by regulating growth of the DRG neuron axons, so that the important effect of microRNA (miRNA) in the nerve injury repair process can be better understood, and a new target is provided for treatment after nerve injury.

Description

technical field [0001] The invention belongs to the field of biomedical technology, and relates to the application of miR-20a in the preparation of drugs for promoting nerve regeneration and repairing nerve damage, in particular to an application of miR-20a in the preparation of drugs for promoting nerve regeneration and repairing nerve damage, miR-20a Application in promoting DRG neuron axon regeneration and peripheral nerve injury repair. Background technique [0002] The repair of peripheral nerve injury is a difficult problem in clinical practice, which brings great burden to the society and family. Compared with the central nervous system, peripheral nerves can spontaneously regenerate after injury, but due to the limited regeneration speed, it is difficult to restore function. Fully and in-depth exploration of the cellular and molecular mechanisms of peripheral nerve injury regeneration will help promote the recovery of peripheral nerve function and provide a theoreti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7088A61P25/00A61P25/02C12Q1/6841C12Q1/6851G01N33/68
CPCA61K31/7088A61P25/00A61P25/02C12Q1/6851C12Q1/6841G01N33/68C12Q2521/107C12Q2531/113C12Q2545/101C12Q2563/107C12Q2543/10C12Q2545/113
Inventor 周松林姚淳赵莉莉于彬顾晓松
Owner NANTONG UNIVERSITY
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