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Application of cecropin polypeptide as anti-inflammatory drug

An anti-inflammatory drug, cecropin technology, applied in the field of biomedicine, can solve problems such as retention, and achieve the effect of low synthesis cost, good application prospects, and small molecular weight

Active Publication Date: 2018-01-19
SUZHOU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, research on the anti-inflammatory physiology and pharmacology of mosquitoes is currently limited to the research level of the sputum complex and the crude extract of the sputum gland, and no specific anti-inflammatory components have been identified, which needs further elaboration

Method used

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  • Application of cecropin polypeptide as anti-inflammatory drug
  • Application of cecropin polypeptide as anti-inflammatory drug
  • Application of cecropin polypeptide as anti-inflammatory drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Aedes aegypti natural immune peptide cecropin inhibits LPS-induced nitric oxide (NO) production in mouse peritoneal macrophages and human peripheral blood mononuclear cells (human PBMCs)

[0047] (1) Inhibit the transcription of inducible nitric oxide synthase (iNOS) in mouse peritoneal macrophages stimulated by LPS

[0048] iNOS is a synthetase necessary for NO production. First, the effect of five cecropins of Aedes aegypti on the transcription level of nitric oxide synthase was tested.

[0049] C57BL / 6 mouse peritoneal macrophages were plated in a 24-well cell culture plate (2.5×10 5 Cells / well), cultured with RMPI-1640 medium (purchased from Gbico, USA) supplemented with 2% fetal bovine serum, 100U / ml ampicillin and 100μg / ml streptomycin sulfate. After the cells adhere to the wall, such as figure 1 As noted, add 100ng / ml lipopolysaccharide (LPS, from Escherichia coli 0111:B4, purchased from Sigma) into the cells, and add 5μM AeaeCec1, AeaeCec2, AeaeCec3, AeaeCec4, AeaeCec5,...

Embodiment 2

[0058] Aedes aegypti natural immune peptide cecropin inhibits lipopolysaccharide (LPS)-induced mouse macrophages (mouseperitoneal macrophages) and human peripheral blood mononuclear cells (human PBMCs) pro-inflammatory cytokine production

[0059] Plate C57BL / 6 mouse peritoneal macrophages or human PBMCs in a 24-well cell culture plate (2.5×10 5 Cells / well), cultured in RMPI-1640 medium (purchased from Gbico, USA) supplemented with 2% fetal bovine serum, 100U / ml ampicillin and 100μg / ml streptomycin sulfate. After the cells adhere to the wall, the cells Add 100ng / ml lipopolysaccharide (LPS, from Escherichia coli 0111:B4, purchased from Sigma), and add 5μM AeaeCec1, AeaeCec2, AeaeCec3, AeaeCec4, AeaeCec5, and the experimental group for synergistic effect detection. AeaeCec1-5 (1 μM each). And set up a control group, add an equal volume of PBS. After a total of 6 hours of incubation, the cell culture supernatant was collected, and the cytokine tumor necrosis factor-α (TNF-α), inter...

Embodiment 3

[0064] Toxicity of Aedes aegypti natural immune polypeptide cecropin to mammalian cells

[0065] C57BL / 6 mouse peritoneal macrophages, human PBMCs and green monkey kidney cell line Vero E6 cells were plated in 96-well culture plates (2×10 4 Cells / well), cultured with RMPI-1640 medium (100μl / well, purchased from Gbico, USA) supplemented with 2% fetal bovine serum, 100U / ml ampicillin and 100μg / ml streptomycin sulfate, and wait for the cells to adhere to the wall After that, a series of peptides diluted in 2 times in serum-free RPMI-1640 medium were added. For 24 hours of co-cultivation, each well was added with cell proliferation and toxicity detection reagent CCK-8 (10μl / well, purchased from Beijing Watbison Technology Co., Ltd. Company), after 4 hours of incubation, detect the light absorption at 450nm, define the cell viability without added polypeptide as 100%, and calculate the cell viability after treatment with different concentrations of polypeptide.

[0066] The results show...

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Abstract

The invention relates to an application of cecropin polypeptide as an anti-inflammatory drug. An amino acid sequence of the cecropin polypeptide is shown as SEQ ID No.1, SEQ ID No.2, SEQ ID No.3, SEQID No.4 or SEQ ID No.5. The invention also discloses the anti-inflammatory drug, comprises the cecropin polypeptide, and the amino acid sequence of the cecropin polypeptide is one or more of SEQ ID No.1-SEQ ID No.5. The invention also discloses five natural immunity polypeptides and the application of cooperative effect of the polypeptides in preparation of the anti-inflammatory drug, and especially relates to the five natural immunity polypeptides sourced from Aedes aegypti and the application of the cooperative effect of the polypeptides.

Description

Technical field [0001] The present invention relates to the field of biomedicine, in particular to the application of cecropin polypeptide as an anti-inflammatory drug. Background technique [0002] Infectious diseases have always been a global public issue threatening human health. Patients infected by pathogenic microorganisms are usually accompanied by uncontrollable inflammation. For example, in the process of bacterial infection, gram-negative bacteria will release a large amount of cell wall component-lipopolysaccharide, and gram-positive bacteria will also release a large amount of cell wall component-lipoteichoic acid, and lipopolysaccharide and lipid Muric acid is an agonist of Toll-like receptor 4 and Toll-like receptor 2, which activates a series of inflammatory signal pathways downstream of immune cells, such as mitogen-activated protein kinase signaling pathway MAPKs and nuclear transcription factor signaling pathway NF- kappa B, etc., resulting in the body over pr...

Claims

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Application Information

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IPC IPC(8): A61K38/17A61P31/04A61P29/00
CPCY02A50/30
Inventor 卫林徐薇杨洋周延东
Owner SUZHOU UNIV
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