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Anti-inflammatory effect of novel anti-inflammatory molecular DRP, implementation method and application

A technology of use and protein, applied in the fields of biotechnology and medicine, can solve problems such as lack of immune function

Inactive Publication Date: 2010-12-01
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although existing studies have revealed the role of DRP / Nrdp1 in regulating cell growth and apoptosis, there is no research on its immune function.

Method used

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  • Anti-inflammatory effect of novel anti-inflammatory molecular DRP, implementation method and application
  • Anti-inflammatory effect of novel anti-inflammatory molecular DRP, implementation method and application
  • Anti-inflammatory effect of novel anti-inflammatory molecular DRP, implementation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0110] Example 1: Inhibitory effect of overexpression of DRP on the production of inflammatory factors in the macrophage cell line RAW264.7

[0111] First, pcDNA3.1 (purchased from Invitrogen) was used as a eukaryotic expression vector, and the cDNA product of DRP with BamHI / KpnI restriction sites was inserted into the corresponding position of the vector, and amplified in E. Coli strain DH-5α The vector is purified to obtain the eukaryotic expression vector of DRP after sequence identification. Stable transfection (transfection reagent JetPei was purchased from Polyplus company) mouse RAW264.7 cells (purchased from ATCC), and then the resulting stable transfected cell line (1 × 10 5 cells / ml RPMI 1640 medium), treated with 100ng / ml LPS (also known as lipopolysaccharide, which is the ligand of TLR4, purchased from Sigma Company) for different time, collected cells and culture supernatant, prepared cDNA for quantitative RT -PCR (94°C, 30Sec; 58°C, 30Sec; 72°C, 30Sec; a total...

Embodiment 2

[0114] Example 2: The inhibitory effect of overexpression of DRP on the production of inflammatory factors in serum and its effect on endotoxic shock Effect on mouse survival

[0115] First, pShuttle2 in the Adeno-X virus vector system (purchased from Clontech Company) was used as the cloning vector, and the cDNA of DRP was inserted between the SalI and XhoI restriction sites, and the shuttle plasmid was recombined with the viral DNA in HEK293 cells, and amplified. After the amplification, the adenovirus purification kit (purchased from Clontech Company) was used. Will 1×10 7 PFU Ad-DRP was injected into the tail vein of mice (6-week-old male SDF-grade C57BL6 mice, purchased from Sipper BK), and LPS (15 mg / kg) was injected intraperitoneally 48 hours later. After 4 hours, the mouse serum was collected by taking blood from the eyeball, and the expression levels of inflammatory factors TNFα, IL-6 and IL-1β in the serum were detected by ELISA method (purchased from R&D Compan...

Embodiment 3

[0118] Example 3: Inhibitory effect of overexpression of DRP on organ damage caused by endotoxic shock

[0119] A replication-deficient adenoviral vector of DRP was constructed (same as Example 2). Will 1×10 7 PFU Ad-DRP was injected into the mouse tail vein (same as Example 2), and LPS (15 mg / kg) was intraperitoneally injected 48 hours later. After 4 hours, the serum of the mice was collected, and the levels of AST and ALT in the serum were detected by a biochemical automatic analyzer (purchased from SRL Company) as liver injury markers. After 24 hours, the liver, spleen and brain of the mice were harvested to observe the gross pathological changes, and the sections were stained with HE to observe the microscopic pathological changes of the liver (focus on necrosis, vacuole formation and inflammatory cell infiltration, etc.).

[0120] The results of the DRP adenovirus vector on the impact of AST and ALT in mouse serum are as follows: Figure 5 shown. The results showed ...

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Abstract

The invention relates to anti-inflammatory effect of novel anti-inflammatory molecular DRP, an implementation method and application, in particular to application of death resistant protein (DRP) in resisting bacterial infection and an application method. The invention provides the application method of the molecule in resisting bacterial infection and a corresponding immunological principle. The invention discloses application and strategy of the molecule in resisting the bacterial infection, particularly in preventing and treating infectious shock and endotoxic shock caused by the bacterial infection. The invention proves that the molecule can inhibit the generation of inflammatory factors after the bacterial infection, improve the survival rate of bodies in the endotoxic shock and protect the infected organs such as liver and the like from inflammation injury.

Description

technical field [0001] The invention belongs to the field of biotechnology and medicine, in particular, the invention relates to the effect, action mechanism, implementation method and application of death resistant protein (DRP) in bacterial infectious diseases. Background technique [0002] Bacterial infection is a common clinical disease, which can lead to local and systemic infectious manifestations of the body, and can even eventually lead to serious consequences such as septic shock, endotoxic shock, and multiple organ dysfunction syndrome (MODS). Once severe infection leads to shock or multiple organ failure, it causes high mortality (~50-80%) performance. Common clinical bacterial infections, such as Escherichia coli, Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Enterococcus faecalis, etc., can cause local and systemic inflammatory reactions in the body, Cause different degrees of infectious diseases. [0003] For p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/17A61K48/00A61P31/04A61P29/00A61P9/00A61P43/00
CPCY02A50/30
Inventor 陈涛涌汪晨曹雪涛
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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