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Application of circular RNA CircOGDH as acute ischemic stroke treatment target

A technology of ischemic stroke and therapeutic target, which is applied in the biological field, can solve the problem that the application of pathological changes in stroke has not been reported, and achieve the effect of reducing neuronal apoptosis

Pending Publication Date: 2022-04-05
THE FIRST AFFILIATED HOSPITAL OF JINAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the correlation between CircOGDH and the pathological changes of acute ischemic stroke and its application have not been reported

Method used

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  • Application of circular RNA CircOGDH as acute ischemic stroke treatment target
  • Application of circular RNA CircOGDH as acute ischemic stroke treatment target
  • Application of circular RNA CircOGDH as acute ischemic stroke treatment target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1. Screening process and identification of CircOGDH

[0022] In the early stage, the joint analysis of next-generation sequencing data of middle cerebral artery occlusion (middle cerebral artery occlusion, MCAO) mouse brain tissue and MCAO mouse blood chip data was carried out, and real-time fluorescent quantitative PCR (Quantitative Real-time PCR, qPCR) was carried out. The verification screen obtained the circular RNA CircOGDH generated by the transcription of the oxoglutarate dehydrogenase (Oxoglutarate Dehydrogenase, OGDH) gene (Fig. 1.A-C). The stability of CircOGDH was further confirmed by RnaseR enzyme treatment (Figure 1.D), and RNA in situ hybridization (FISH) was used to find that CircOGDH was mainly expressed in the cytoplasm of ischemic penumbra neurons in MCAO mice (Figure 1.E) .

Embodiment 2

[0023] Example 2. Knocking down CircOGDH can reduce the apoptosis of primary neurons caused by ischemia and hypoxia

[0024] 2.1 Transfection of sh_CircOGDH adenovirus can knock down the expression of CircOGDH in primary neurons

[0025] In order to explore the biological function of CircOGDH in the process of hypoxia-induced neuronal apoptosis, we designed the ADV_GFP_shRNA_CircOGDH adenovirus (sh_CircOGDH for short) targeting the reverse splicing sequence of CircOGDH to knock down the expression of CircOGDH in cells ( Figure 2A ). Transfection of ADV_GFP_shRNA_CircOGDH adenovirus or ADV_GFP_shRNA_CircCtrl adenovirus (negative control) in primary neurons, high expression of GFP (green fluorescence) can be observed under a fluorescent microscope, proving that adenoviruses can invade primary neuronal cells ( Figure 2B ). Through RT-qPCR experiments, we found that transfection of sh_CircOGDH adenovirus could significantly reduce the expression of CircOGDH in primary neuronal ...

Embodiment 3

[0029] Example 3. Knocking down CircOGDH reduces neuronal apoptosis in MCAO model mice

[0030] 3.1 Experimental Design and Construction of MCAO Model Mice

[0031] Experimental design such as Figure 4A As shown, use a stereotaxic apparatus to inject sh_CircCtrl or sh_CircOGDH adenovirus into two parts of the left cortex of the mouse, and the specific coordinates are as follows: 1) 0.2 mm behind the Bregma point, 2 mm left, and 1.5 mm deep; 2) 0.2 mm behind the Bregma point, left 3.0mm, 2.0mm deep. On the 6th day after virus injection, MCAO modeling was carried out ( Figure 4A ), 3 hours after modeling, samples were taken for follow-up experiments. Since the adenovirus carries the GFP fluorescent protein, in order to determine whether the virus injection site is accurate and the spread of the virus, we injected the sh_CircCtrl adenovirus into the cortex of MCAO mice and took samples, and made frozen sections of the brain tissue, and then performed immunofluorescence exper...

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Abstract

The invention provides application of circular RNA CircOGDH as a therapeutic target of acute ischemic stroke, and particularly relates to application of circular RNA CircOGDH to preparation of molecular targeting drugs for preventing and / or treating acute ischemic stroke and application of circular RNA CircOGDH to preparation of neuroprotective agents for acute ischemic stroke. The circular RNA CircOGDH disclosed by the invention is highly expressed in ischemic and anoxic neurons after the acute ischemic stroke is attacked, and neuronal apoptosis can be relieved by knocking down the CircOGDH, so that the CircOGDH is a potential protective target of neuronal apoptosis after the acute ischemic stroke is attacked. The invention provides a new target for developing acute ischemic stroke neuroprotective drugs.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of a circular RNA CircOGDH as a therapeutic target for acute ischemic stroke. Background technique [0002] Acute ischemic stroke (AIS), also known as acute cerebral infarction (AIS), refers to the necrosis of brain tissue caused by the sudden interruption of cerebral blood supply. It has a high incidence and is a disease of human death and disability. main cause. The key to the treatment of acute cerebral infarction is to open the blocked blood vessels as soon as possible and save the ischemic penumbra. At present, intravenous thrombolysis and arterial thrombectomy are the main treatment methods for acute cerebral infarction. The earlier the treatment time, the more benefits. However, both intravenous thrombolysis and arterial thrombectomy have time windows, and patients who exceed the time window will die. These two effective treatments were unacceptab...

Claims

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Application Information

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IPC IPC(8): A61K45/00A61P9/10A61P25/00
Inventor 逯丹徐安定臧健坤苏璇琳李玉峰刘燕芳张添源吴有盛曾智军李克深谭泽锋王世勇
Owner THE FIRST AFFILIATED HOSPITAL OF JINAN UNIV
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