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Application of gene NOVA1 in preparation of medicine for treating nerve cell hypoxia injury

A NOVA1, nerve cell technology, applied in the field of medical biochemistry, can solve problems such as unreported research

Inactive Publication Date: 2015-04-29
YANGZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Nova1, as a nerve-specific splicing factor, is often used to study its alternative splicing regulation network, but the study in hypoxia has not been reported

Method used

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  • Application of gene NOVA1 in preparation of medicine for treating nerve cell hypoxia injury
  • Application of gene NOVA1 in preparation of medicine for treating nerve cell hypoxia injury
  • Application of gene NOVA1 in preparation of medicine for treating nerve cell hypoxia injury

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Construction of Nova-1's Eukaryotic Expression Vector

[0020] We purchased the gene Homo sapiens neuro-oncological ventral antigen 1 (NOVA1), transcript variant 1 NM_002515 BC075038 from Hunan Changsha Yingrun Biotechnology Company, and performed PCR amplification using the cloning vector pCR4-TOPO as a template. See figure 1 The nova1 was amplified by PCR to get a clear band, and Nova1 of 1500bp was obtained (1, 3, DL2000 Marker; 2, 15μl pcr product; 4,7.5μl pcr product). The gel was then recovered, digested with salI and xhoI, and then re-recovered, ligated with the eukaryotic expression vector pcmv-myc digested with salI and xhoI, and transferred to DH5α for screening. And perform restriction enzyme digestion to identify the recombinant plasmid, see figure 2 The recombinant plasmid pcmv-myc-Nova1 was identified by restriction digestion: 1 hole was DL2000 Marker, 2 hole was pcmv-myc-Nova1 digested by salⅠand xhoⅠ, and 3 hole was pcmv-myc digested by salⅠan...

Embodiment 2

[0021] Example 2 The eukaryotic expression vector pCMV-Myc-Nova1 was transfected into PC12 cells.

[0022] (1) Detection of cytotoxicity of Lipo-2000 transfected PC12.

[0023] The PC12 cells were transfected with Lipo-2000 to form a Lipo-2000 transfection group, and the untransfected PC12 cell group was set as a reference.

[0024] The untransfected group and the Lipo-2000 transfected group, each with 5 wells, were used to detect the survival rate of cells at different times by the MTT method, repeated 3 times, the test results showed that the Lipo-2000 transfected group was transfected at 24h, 48h, 72h And after 96h, the cell survival rate did not decrease significantly compared with the untransfected group.

[0025] (2) The vector pCMV-Myc-Nova1 was transfected into PC12 cells.

[0026] PC12 cells were transfected with pCMV-Myc-Nova1 to form a pCMV-Myc-Nova1 group, an untransfected group, and a Pcmv-myc vector and empty vector group without nova1 as a reference.

[0027] (1) Scree...

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Abstract

The invention relates to the field of medical biochemistry and in particular relates to a medicine for treating nerve cell hypoxia injury. A Nova1 hypoxia injury protective effect is explored by constructing a Nova1 eukaryotic expression vector and simulating neuron cell hypoxia conditions. A stable physical hypoxia injury model is built by taking PC12 cells, namely rat adrenal pheochromocytoma, as an object; the Nova1 eukaryotic expression vector Pcmv-myc-Nova1 is constructed by adopting digestion and PCR methods, and sequencing verification is carried out; and a cell transfection experiment is carried out, cellular morphology, RNA and protein level are detected, detection results find that a nerve specificity splicing factor Nova1 has a hypoxia injury protective effect, and a foundation is laid for research of a hypoxia injury protection mechanism in future.

Description

technical field [0001] The invention relates to the field of medical biochemistry, in particular to a medicine for treating nerve cell hypoxic injury. Background technique [0002] The brain is the organ with the most active energy metabolism in the body, with the largest blood flow and oxygen consumption. Cerebral stroke has become one of the major diseases that cause death and disability in the world today, but its mechanism has not been fully elucidated. Oxygen free radical damage, lipid peroxidation, inflammatory cytokines, and nitric oxide-mediated damage were studied; in terms of treatment, measures such as scavenging free radicals, reducing calcium overload, and application of cytoprotective agents were used, but the overall curative effect was poor. micro. The brain is the most complex organ in the body, and the neural-specific splicing factors and their regulation are obviously closely related to the physiological and pathological states of the brain. Nova1, as a ...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P25/00
Inventor 李华玲孔玲吕贝夏靖陈欣虹
Owner YANGZHOU UNIV
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