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Application of Sirt1 in repairing corneal nerve injury

A technology for nerve injury and cornea, which is applied in the application field of silent signal regulator 1 in the repair of corneal nerve injury, and can solve the problems of corneal nerve regeneration and corneal nerve injury repair with little effect.

Inactive Publication Date: 2016-09-28
QINGDAO CENT HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In recent years, surgical methods such as amniotic membrane transplantation and corneal limbal stem cell transplantation have appeared to promote the healing of persistent corneal epithelial defects. The problem

Method used

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  • Application of Sirt1 in repairing corneal nerve injury
  • Application of Sirt1 in repairing corneal nerve injury
  • Application of Sirt1 in repairing corneal nerve injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1: Sirt1 Promotes Axonal Growth of Mouse Trigeminal Ganglion Cells

[0028] The culture medium of trigeminal ganglion cells ( Medium, Invitrogen / Gibco, Cat. No. 21103-049), plus B27 (10X) (Invitrogen / Gibco, Cat. No. 17504-044). Routine culture of trigeminal ganglion cells, 1000 cells were seeded into 30mm culture dishes, and then in 5% CO 2 Cultured at 37°C, replaced with new medium every 3 days, and added Sirt1 factor at the same time, fixed the cells after 9 days of culture, and stained with neuron-specific marker Tubulin III to observe the axonal growth of trigeminal ganglion cells. Such as figure 1 As shown, after adding 20-50ng / ml Sirt1 to the culture medium, the length of the axon of the trigeminal ganglion cell was significantly increased, and the branch of the axon was significantly increased, indicating that Sirt1 can effectively promote the growth of the axon of the trigeminal ganglion cell.

Embodiment 2

[0029] Example 2: The combination of Sirt1 and miR-182-5p agonist promotes axonal growth of mouse trigeminal ganglion cells

[0030] The culture medium of trigeminal ganglion cells ( Medium, Invitrogen / Gibco, Cat. No. 21103-049), plus B27 (10X) (Invitrogen / Gibco, Cat. No. 17504-044). Routine culture of trigeminal ganglion cells, 1000 cells were seeded into 30mm culture dish, Sirt1 (20ng / ml) was added to the dish, and then in 5% CO 2 Cultured at 37°C, replaced with new medium every 3 days, added miR-182-5p agonist at a concentration of 0.1 or 0.5nmol / ml, fixed cells after 9 days of culture, and stained with neuron-specific marker Tubulin III , to observe the axonal growth of trigeminal ganglion cells. Such as figure 2 As shown, after adding 0.1 or 0.5 nmol / ml miR-182-5p agonist to the culture medium, the axon length of trigeminal ganglion cells was significantly increased, and the axon branch was significantly increased, indicating that Sirt1 and miR-182-5p stimulated The...

Embodiment 3

[0031] Example 3: The effect of exogenous addition of Sirt1 on the repair of corneal nerve damage in normal mice

[0032] Twelve C57BL / 6 mice (6-8 weeks old) were randomly divided into control group and experimental group, and the epithelium in the central 3mm area of ​​the right eye cornea of ​​the mice was scraped using a 3mm trephine drill and an epithelial scraper. 5 μl PBS was injected subconjunctivally in the right eye, and 50 ng Sirt1 (dissolved in 5 μl PBS solution) was injected subconjunctivally in the right eye of the mice in the experimental group, and the neuron-specific marker Tubulin III staining was used to observe the expression level of mice in each group 21 days after modeling. Corneal Nerve Regeneration, representative photographs see image 3 . Such as image 3As shown, subconjunctival injection of 50ng Sirt1 can effectively promote the regeneration of corneal nerves.

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Abstract

The invention provides application of Sirt1 in repairing corneal nerve injury. It is discovered herein that Sirt1 may promote the repairing of corneal nerve injury and may also promote the preparing of corneal epithelial injury as well as the axonal growth of trigeminal ganglion cells; the Sirt1 may be used to culture trigeminal ganglion cells, repair persistent corneal epithelial defect, repair diabetic corneal nerve injury and repair nervous keratitis. Moreover, corneal nerve injury can be more effectively repaired by the joint use of silencing signal regulator 1 and miR-182-5p agonist.

Description

technical field [0001] The invention belongs to the technical field of ophthalmic disease treatment, and in particular relates to the application of silencing signal regulator 1 (Sirt1) in repairing corneal nerve damage. Background technique [0002] The cornea is an avascular, transparent tissue whose nutrients mainly come from tears, aqueous humor, and nerve fibers. The cornea is rich in nerve fibers and is one of the most nerve-dense tissues in the human body. It has a keen sense of cold, heat, pain, touch and pressure. The corneal nerve originates from the ophthalmic branch of the trigeminal nerve and plays an important role in maintaining the corneal tissue, especially the function and anatomical integrity of the corneal epithelium. Corneal nerve injury will cause corneal sensory loss, resulting in neurotrophic corneal epithelial disease, the main manifestations include repeated epithelial erosion, slow epithelial regeneration, persistent epithelial defect, ulceration,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K45/06A61P25/02A61P27/02A61P3/10C12N5/079
CPCA61K38/1709A61K45/06C12N5/0618C12N2501/00C12N2501/65
Inventor 王晔牟晓峰张蓉慕莹
Owner QINGDAO CENT HOSPITAL
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