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Application of circular RNA circ-Ankib1 in preparation of medicament for promoting nerve regeneration and repairing nerve injury

A technology for nerve injury and nerve regeneration, applied in the field of nerve regeneration and repair

Active Publication Date: 2019-08-20
NANTONG UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the role of circRNAs in peripheral nervous system injury repair has been rarely reported so far, especially the phenotype regulation of Schwann cells

Method used

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  • Application of circular RNA circ-Ankib1 in preparation of medicament for promoting nerve regeneration and repairing nerve injury
  • Application of circular RNA circ-Ankib1 in preparation of medicament for promoting nerve regeneration and repairing nerve injury
  • Application of circular RNA circ-Ankib1 in preparation of medicament for promoting nerve regeneration and repairing nerve injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Screening and identification of circ-Ankib1

[0030] RNA-seq was performed on the sciatic nerves of SD (Sprague Dawley) rats at different time points (0d, 1d, 4d, 7d, 14d) after sciatic nerve crush. According to the sequencing results, select circRNAs that are all composed of exons and have significantly different expressions of loops >500nt. The sequencing results analyzed the screened circRNAs, and further verified the existence of circRNAs by PCR. According to the design principles of circRNAs primers, back primers were designed for these circRNAs, and NCBI was used to design primers for each circRNA. Select different annealing temperatures according to the primer information for PCR amplification, and then select circRNAs with the expected band size through agarose gel electrophoresis, send them to the company for sequencing, and compare the sequencing results with the original sequence to verify whether they span the circularization location. Finally,...

Embodiment 2

[0057] Example 2: Functional verification of circRNA

[0058] 1. Culture of primary Schwann cells

[0059] Take out the sciatic nerve of a 1-day-old SD rat, cut it into pieces with micro scissors, add 1ml of 3mg / ml collagenase, digest at 37°C for 30min, shake once every 10min; centrifuge at 1200rpm for 5min at room temperature, discard the collagenase, and add trypsin 1ml, placed in a 37°C incubator for about 8 minutes, added 3ml of complete medium to stop digestion, passed through a 200-mesh sieve, 1200rpm, and centrifuged at room temperature for 5min; then added 2ml of complete medium and washed 1-2 times; Cultured in a culture dish coated with poly-L-lysine (PLL) (5% CO 2 , 37℃); on the second day, change to the complete medium containing Ara-c (10μM) to inhibit the rapid proliferation of fibroblasts; on the fourth day, change to medium containing HRG (50ng / ml) and Forskolin (2μM) Complete medium stimulates the growth of cells. After the cells grow to a density above 90%...

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Abstract

The invention discloses an application of circular RNA circ-Ankib1 in the preparation of a medicament for promoting nerve regeneration and repairing nerve injury. The result shows that the down-regulation or inhibition of the expression of circ-Ankib1 can promote the proliferation of Schwann cells, and is beneficial to the repair of peripheral nerve injury.

Description

technical field [0001] The invention belongs to the technical field of nerve regeneration and repair, and in particular relates to the application of circular RNA circ-Ankib1 in the preparation of drugs for promoting nerve regeneration and repairing nerve damage. Background technique [0002] Peripheral nerve injuries are usually caused by traumatic injuries such as stretching, ischemic, penetrating, and crushing. Unlike the central nervous system, axons in the peripheral nervous system have some intrinsic regeneration capacity after injury. Schwann cells are important glial cells in the peripheral nervous system and play an important role in the nervous system. After peripheral nerve injury, Schwann cells proliferate and migrate to form cell cords, and at the same time provide a suitable microenvironment for nerve regeneration after injury by synthesizing and secreting neurotrophic factors and activating immune responses. Studying the key factors and pathways in the proce...

Claims

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Application Information

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IPC IPC(8): A61K31/7105A61P25/00A61P25/02
CPCA61K31/7105A61P25/00A61P25/02
Inventor 于彬姚淳毛苏苏张珊珊顾晓松
Owner NANTONG UNIVERSITY
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