Pharmaceutical application of lentivirus in regulating sulfydryl nitrosylation modification level of Septin2

A nitrosylation and horizontal technology, applied in gene therapy, antineoplastic drugs, drug combinations, etc., to achieve the effect of inhibiting the occurrence of aortic aneurysm and aortic dissection

Active Publication Date: 2021-07-09
NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, whether it can undergo sulfhydryl nitrosylation modification and its ro

Method used

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  • Pharmaceutical application of lentivirus in regulating sulfydryl nitrosylation modification level of Septin2
  • Pharmaceutical application of lentivirus in regulating sulfydryl nitrosylation modification level of Septin2
  • Pharmaceutical application of lentivirus in regulating sulfydryl nitrosylation modification level of Septin2

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Experimental program
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Embodiment 1

[0018] 1. Biotin-Switch method to detect thiol-nitrosylation-modified proteins (Cayman Chemical Company S-nitrosylation Protein Detection Kit, Cat. No. 10006518)

[0019] (1) After the cells are treated, remove the medium, wash three times with pre-cooled wash buffer, add 500 μL washbuffer to scrape, and transfer to a 1.5 mL centrifuge tube.

[0020] (2) Centrifuge at 500×g at 4°C for 5 min, and discard the supernatant.

[0021] (3) Place the cells on ice and configure the Blocking buffer. The specific preparation method is as follows: 100 μL of DMF and 900 μL of buffer A are added to the new blocking reagent to form the blocking buffer stock solution. After mixing, add 9 mL of buffer A, which is the Blocking buffer.

[0022] (4) Add 500 μL blocking buffer to each tube to resuspend the cell pellet obtained in (2), and rotate at 4°C for 30 min.

[0023] (5) Centrifuge at 12,000 rpm for 10 min at 4°C, and transfer the supernatant to a 15 mL centrifuge tube.

[0024] (6) Add 4 ...

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Abstract

The invention relates to a pharmaceutical application of a lentivirus in regulating the sulfydryl nitrosation modification level of Septin2, belonging to the field of molecular biology diagnosis of aortic aneurysm and aortic dissection. According to the invention, the occurrence of the aortic aneurysm and the aortic dissection can be effectively inhibited by infecting the lentivirus into macrophages to down-regulate the sulfydryl nitrosation modification of the Cys111 site of the Septin2 protein.

Description

technical field [0001] The invention belongs to the field of molecular biology diagnosis of aortic aneurysm and aortic dissection, in particular to the pharmaceutical use of regulating the level of Septin2 sulfhydryl nitrosylation. Background technique [0002] Aortic aneurysm and aortic dissection (AAD) is a life-threatening disease with a high mortality rate in the event of aortic rupture. Over the past 20 years, major advances have been made in open surgery and endovascular repair of AAD. However, drug therapies are still lacking, therefore, the molecular and cellular mechanisms involved in the formation and development of AAD need to be explored. The main pathological features of AAD are the loss of vascular smooth muscle cells and the remodeling of the extracellular matrix due to the accumulation and activation of inflammatory cells. Inflammatory responses play a crucial role in AAD and largely influence many determinants of aortic wall remodeling. Macrophage-mediate...

Claims

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Application Information

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IPC IPC(8): A61K48/00A61P9/00A61P35/00G01N27/62G01N33/574G01N33/68
CPCG01N33/6893G01N33/57407G01N33/57492G01N33/57496A61K48/0008A61K48/005A61P35/00A61P9/00G01N27/62G01N2800/329G01N2333/47
Inventor 季勇张昊张艳张旭红严科韩艺谢利平
Owner NANJING MEDICAL UNIV
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