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Application of CART to preparation of ischemic brain injury neuroprotective agent medicine

A technology of ischemic brain injury and neuroprotective agent, which is applied in the application field of preparing neuroprotective agent for ischemic brain injury, can solve problems such as lack, and achieve the effects of increasing expression, promoting neuron regeneration, and reducing infarct volume

Pending Publication Date: 2021-12-03
NANJING DRUM TOWER HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is still a lack of such brain-protective drugs in clinical practice.

Method used

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  • Application of CART to preparation of ischemic brain injury neuroprotective agent medicine
  • Application of CART to preparation of ischemic brain injury neuroprotective agent medicine
  • Application of CART to preparation of ischemic brain injury neuroprotective agent medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] Example 1 In vitro study of the therapeutic effect of CART on ischemic stroke neurons

[0022] Culture of primary cortical neurons: Pregnant 15-17d wild-type Kunming mice were killed by dislocation of the neck, and the embryos were collected and placed in a petri dish filled with cold HBSS, the cerebral cortex was separated, digested with 1X trypsin, and placed in a polymer Cultured in 96-well culture plate after lysine treatment, 2~3×10 6 Cells / mL, change medium every 2-3d, co-culture for 10-14d; cell culture at 37℃, 5% CO 2 environment in the incubator.

[0023] Drug treatment: Oxygen-glucose deprivation (OGD) neuron model was prepared by primary cultured neuron cells, and CART (0.2, 0.4, 0.8, 1.6nM) was added for another 24 hours, and normal saline (Saline) was added to OGD as a control group In the experiment, the dose of CART was finally selected as 0.4nM.

[0024] Detection of neuron cell viability: CCK-8 method was used to detect the cell viability of neurons;...

Embodiment 2

[0027] Example 2 In vivo study of the therapeutic effect of CART on brain tissue of ischemic stroke

[0028] Mouse middle cerebral artery occlusion (MCAO)-reperfusion (I / R) model establishment: male C57BL / 6J (B6) mice (about 2 months old, body weight 25±2g) were selected. Mice were anesthetized by intraperitoneal injection of sodium pentobarbital (1%) at a dose of 45 mg / kg. During surgery, the body temperature of the mice was monitored and maintained at 37.0 °C. A 6-0 surgical monofilament nylon suture was inserted into the internal carotid artery through the stump of the external carotid artery, and one side of the internal carotid artery was blocked to induce transient focal cerebral ischemia.

[0029] Drug treatment: 2.5 μg / kg / time of CART55-102 tail vein injection (with normal saline (Saline) group as the control) was given before the start of ischemia 2h reperfusion, and the administration was repeated once every 24 hours. At 24h, 48h, 72h and 1 week after reperfusion, ...

Embodiment 3

[0031] Example 3 The multi-target effect of CART on the neuroprotective effect of the main pathological process of ischemic stroke

[0032] Observation of synaptic ultrastructure under a transmission electron microscope: The brain was taken from the mouse after 24 hours of ischemia and reperfusion, and the complete brain tissue was taken out, the cortex was separated on ice, and the isolated brain tissue was trimmed to less than 1cm 3 The pieces were fixed overnight in 2.5% glutaraldehyde + 4% paraformaldehyde. Gradient dehydration, embedding and solidification were carried out again, and slices were made with an ultrathin microtome to prepare brain tissue slices with a thickness of about 50 nm, which were double-stained with 3% uranyl acetate-lead citrate; the number of synapses, Synaptic vesicle and synaptic cleft widths within the presynaptic membrane. TEM data were processed with Photoshop software, and ImagJ software was used for statistics.

[0033] Determination of ne...

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Abstract

The invention relates to an application of CART to preparation of ischemic brain injury neuroprotective agent medicine, and the CART is prepared into an injection preparation. At present, an effective neuroprotective agent for cerebral arterial thrombosis does not exist clinically, CART is one of few peptide substances which are found at present and can pass through a blood-brain barrier as an endogenous small-molecule neuropeptide, the CART firstly regulates and controls the synaptic plasticity of ischemic neurons, the protein gene expression is regulated and controlled by regulating and controlling cytoskeleton activity, the ischemic neuron synapsin expression and the number of synapses are increased, and neuron axon growth is promoted; secondly, neuronal regeneration is promoted, nerve cell apoptosis is reduced, and ischemic brain injury is relieved. The CART not only can regulate and control the synaptic plasticity of ischemic neurons, but also can promote the regeneration of neurons, and simultaneously acts on a plurality of targets in the pathological process of ischemic brain injury, so that the dual effects exactly accord with the current treatment strategy of cerebral arterial thrombosis.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to the application of CART as a medicine for preparing neuroprotective agent for ischemic brain injury. Background technique [0002] Stroke is one of the three major diseases that threaten human health, about 85% of which are ischemic strokes, which are characterized by high morbidity, high mortality, high disability and high recurrence rate, and are increasingly younger; More than 70% of the survivors have different degrees of dysfunction. The pathological mechanism of ischemic brain injury is complex. In recent years, a large number of studies have been carried out on the mechanism of neuron injury induced by cerebral ischemia, and a series of theories of neuron injury and death have been established. Based on these theories, most neuron injury Although neuroprotective agents have observed good effects in animal experiments, they have not been successfully used clinicall...

Claims

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Application Information

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IPC IPC(8): A61K38/22A61P9/10A61P25/00
CPCA61K38/2271A61P9/10A61P25/00
Inventor 沙杜鹃
Owner NANJING DRUM TOWER HOSPITAL
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