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Tool for diagnosing glaucoma and drug target for treating glaucoma

A glaucoma and tool technology, applied in the direction of drug combination, pharmaceutical formula, microbial measurement/inspection, etc., can solve problems such as visual function impairment

Active Publication Date: 2020-03-13
广州雏鸟信息科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these tests can diagnose glaucoma, studies have shown that more than 50% of patients with glaucoma diagnosed by morphological and functional means have impaired visual function

Method used

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  • Tool for diagnosing glaucoma and drug target for treating glaucoma
  • Tool for diagnosing glaucoma and drug target for treating glaucoma
  • Tool for diagnosing glaucoma and drug target for treating glaucoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1 Screening of genes differentially expressed in glaucoma patients and normal population

[0055] 1. Sample collection

[0056] During glaucoma surgery, 50 cases of lesion optic nerve head tissue and 45 cases of normal traumatized optic nerve head tissue were collected as controls. All glaucoma patients and their families signed the informed consent.

[0057] 2. Total RNA extraction

[0058] Extracted with TRIzol (Invitrogen) reagent, and stored at -80°C for future use. After adding TRIzol, place it at room temperature for 10 minutes to fully lyse the sample (note: if the next step is not performed, the sample can be stored at -70°C for long-term storage). Add 200 μl of chloroform to every 1ml of TRIzol, shake vigorously and mix well, then place at room temperature for 3-5min to allow natural phase separation. Centrifuge at 12,000 rpm at 4°C for 15 min. The sample will separate into three layers: a yellow organic phase, an intermediate layer and a colorless...

Embodiment 2

[0082] The therapeutic efficacy detection of embodiment 2HCK

[0083] 1. Culture of RGC-5 cell line

[0084] Resuscitate the RGC-5 cells, take the RGC-5 cell line out of the liquid nitrogen tank, and immediately put it in a water bath at 36°C for 1 min, and when there is a small amount of ice, shake the cryovial gently to melt the ice. Centrifuge at 1000r / min for 5min, suck out the supernatant, add DMEM containing 10% serum to resuspend the cells, adjust the density and inoculate in a clean culture bottle. Placed in 95% air, 37°C, 5% CO 2 The cells were cultured in a cell incubator with saturated humidity, and subsequent experiments were performed when the cells reached 70% confluency.

[0085] 2. Interfering RNA design and synthesis

[0086] siRNA against HCK (HCK-siRNA)

[0087] Sense strand: 5'-AUCUUGUAAUGUUUCACGGUATT-3' (SEQ ID NO.5);

[0088] Antisense strand: 5'-CCGUGAAACAUUACAAGAUCCTT-3' (SEQ ID NO. 6).

[0089] The above siRNA was synthesized by Shanghai Gemma Ph...

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Abstract

The invention discloses a tool for diagnosing glaucoma and a drug target for treating the glaucoma. According to the invention, HCK gene expression of a glaucoma patient is proved to be significantlyup-regulated; a glaucoma diagnosis tool is developed by utilizing HCK to distinguish the nature of the glaucoma patient; and the tool can be a kit, a chip and the like. Retinal ganglion cells are cultured in vitro; the damage state of the retinal ganglion cells can be reversed by interfering in the HCK, which indicates the HCK can be used for developing a medicine for the glaucoma; and the medicine achieves the purpose of treating the glaucoma by adjusting the expression level of the HCK.

Description

technical field [0001] The invention relates to the field of diagnosis and treatment of glaucoma, more specifically, the invention relates to a tool for diagnosing glaucoma and a drug target for treating glaucoma. Background technique [0002] Glaucoma is a common and difficult eye disease with intermittent or continuous elevation of intraocular pressure. The onset of the disease is rapid, harmful, and can cause blindness at any time. Sustained high intraocular pressure can cause damage to various parts of the eyeball and visual function, leading to optic atrophy, narrowed field of vision, and decreased vision. Blindness is only a matter of time. Complete blindness can occur within 24-48 hours of the acute attack. If left untreated, the visual field can be completely lost and even blind. Glaucoma is one of the three major blindness diseases that cause blindness in humans, with an incidence rate of 1% in the general population and 2% after the age of 45. Glaucoma is one o...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883A61K45/00A61P27/06
CPCC12Q1/6883A61K45/00A61P27/06C12Q2600/158
Inventor 李成芳马红梅
Owner 广州雏鸟信息科技有限公司
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