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Application of plasma exosome CircOGDH as diagnostic biomarker of acute ischemic stroke

A technology of ischemic stroke and biomarkers, applied in the biological field, can solve problems such as the lack of biomarkers to assist diagnosis

Active Publication Date: 2022-03-15
THE FIRST AFFILIATED HOSPITAL OF JINAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of the lack of biomarkers to assist in the diagnosis of acute ischemic stroke, the present invention proposes the application of plasma exosomal CircOGDH as a diagnostic biomarker for acute ischemic stroke

Method used

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  • Application of plasma exosome CircOGDH as diagnostic biomarker of acute ischemic stroke
  • Application of plasma exosome CircOGDH as diagnostic biomarker of acute ischemic stroke
  • Application of plasma exosome CircOGDH as diagnostic biomarker of acute ischemic stroke

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] Example 1. Ischemic-hypoxic neurons transport CircOGDH to peripheral blood through exosomes

[0032] In order to verify the scientific hypothesis of "whether CircOGDH can be transported from ischemic brain tissue to peripheral blood through exosomes", we constructed an adenovirus-packaged shRNA that targets the back-splicing site of CircOGDH to knock down the expression of CircOGDH , and carry GFP fluorescent protein, the English abbreviation is ADV_GFP_shRNA_CircOGDH, referred to as sh_CircOGDH; the English abbreviation of the negative control (negative control, NC) adenovirus is ADV_GFP_shRNA_NC, referred to as sh_CircCtrl, and it has been verified by experiments that the sh_circOGDH adenovirus can knock down CircOGDH in mouse brain tissue. In addition, we further designed the following experiment: the mice were divided into SHAM (sham operation)+sh_CircCtrl group, middle cerebral artery occlusion model (MCAO)+sh_CircCtrl group and MCAO+sh_CircOGDH group, and mice brai...

Embodiment 2

[0033] Example 2. High expression of exosomal CircOGDH in ischemic hypoxic neurons

[0034] 2.1 Extraction and identification of primary neuronal exosomes

[0035] After the primary neurons were cultured to the fifth day, they were divided into normoxia group and ischemia-hypoxia-reperfusion (anoxia-ischemia(AI) / reperfusion) group for modeling. Purification of exosomes from primary neuronal culture medium. Transmission electron microscopy showed that the average size of exosomes collected from primary neuron culture medium was about 100nm ( Figure 2a ). The size of the primary neuronal exosomes of the two groups was further confirmed by Nanoparticle Tracking Analysis (NTA). 150nm, basically consistent with the particle size of exosomes ( Figure 2b ). Western blot experiments confirmed that CD63, TSG101 and HSP70 exosomal protein markers existed in primary neuronal exosomes ( Figure 2c ).

[0036] 2.2 High expression of CircOGDH in primary neuronal exosomes after isch...

Embodiment 3

[0038] Example 3. High expression of CircOGDH in exosome plasma from patients with acute ischemic stroke (AIS)

[0039] 3.1 Extraction and identification of plasma exosomes

[0040] We purified exosomes from the plasma of non-cerebrovascular disease controls (NCD) subjects using a polymer precipitation method. Transmission electron microscopy showed that the average size of exosomes collected from patient plasma was about 100 nm ( Figure 4a ). It was further confirmed by nanoparticle tracking analysis (NTA) that the size of exosomes in the plasma of subjects in the NCD control group, the particle size corresponding to the peak value of the NCD control group was 126nm, which basically conformed to the size of the exosomes ( Figure 4b ).

[0041] 3.2 CircOGDH expression level increased in plasma exosomes of AIS patients

[0042] We extracted exosomes from the plasma of the research subjects and detected the expression of CircOGDH in the exosomes, including 31 cases in the ...

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Abstract

The invention provides an application of plasma exosome CircOGDH as a diagnostic biomarker of acute ischemic stroke, and particularly relates to an application of plasma exosome CircOGDH as a diagnostic biomarker in preparation of a detection reagent, a diagnostic kit or a diagnostic preparation of acute ischemic stroke. The circular RNA CircOGDH disclosed by the invention is highly expressed in ischemic and hypoxic neurons after the acute ischemic stroke is attacked, and is transferred to peripheral blood from brain tissues through exosomes, so that the expression of the exosomes CircOGDH in plasma is increased, and the circular RNA CircOGDH can be used as a diagnostic biomarker for detecting the acute ischemic stroke. In addition, the plasma exosome CircOGDH disclosed by the invention, as a biomarker for diagnosing the AIS, has relatively high sensitivity and specificity and has a clinical popularization value.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of plasma exosome CircOGDH as a diagnostic biomarker for acute ischemic stroke. Background technique [0002] Acute ischemic stroke (AIS), also known as cerebral infarction, refers to local brain tissue ischemia and hypoxic necrosis caused by sudden interruption of blood supply to the brain due to various reasons, resulting in corresponding neurological dysfunction. A class of clinical syndromes with defects, which have a high incidence rate and are the main cause of human death and disability. The key to the treatment of acute ischemic stroke 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883C12N15/11
CPCC12Q1/6883C12Q2600/158C12Q2600/178C12Q2600/166
Inventor 徐安定逯丹刘燕芳张添源李玉峰臧健坤吴有盛曾智军李克深黄立安张玉生谭泽锋王世勇黎耀杰杨振国
Owner THE FIRST AFFILIATED HOSPITAL OF JINAN UNIV
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