The application of cecropin polypeptide as an anti-inflammatory drug

The technology of an anti-inflammatory drug and cecropin, which is applied in the field of biomedicine, can solve problems such as staying, and achieve the effects of low synthesis cost, strong anti-inflammatory effect and good application prospect.

Active Publication Date: 2020-10-09
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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  • The application of cecropin polypeptide as an anti-inflammatory drug
  • The application of cecropin polypeptide as an anti-inflammatory drug
  • The application of cecropin polypeptide as an anti-inflammatory drug

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Aedes aegypti natural immune polypeptide cecropin inhibits LPS-induced nitric oxide (NO) production in mouse peritoneal macrophages and 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. Firstly, the effect of five cecropins in Aedes aegypti on the transcription level of nitric oxide synthase was detected.

[0049] C57BL / 6 mouse peritoneal macrophages were plated in 24-well cell culture plates (2.5×10 5 cells / well), cultured with RMPI-1640 medium (purchased from U.S. Gbico Company) added with 2% fetal bovine serum, 100 U / ml ampicillin and 100 μg / ml streptomycin sulfate, after the cells adhered, as figure 1 As marked, add 100ng / ml lipopolysaccharide (LPS, from Escherichia coli 0111:B4, purchased from Sigma) to the cells, and add 5μM AeaeCec1, AeaeCec2, AeaeCec3, AeaeCec4, AeaeCec5 respectively for synergistic effect Aeae...

Embodiment 2

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

[0059] C57BL / 6 mouse peritoneal macrophages or human PBMCs were plated in 24-well cell culture plates (2.5×10 5Cells / well), cultivated with the RMPI-1640 medium (purchased from U.S. Gbico Company) that was added with 2% fetal bovine serum, 100 U / ml ampicillin and 100 μg / ml streptomycin sulfate. Add 100ng / ml lipopolysaccharide (LPS, from E. coli Escherichia coli 0111:B4, purchased from Sigma), and add 5μM AeaeCec1, AeaeCec2, AeaeCec3, AeaeCec4, AeaeCec5 respectively, and the experimental group for synergistic effect detection was added at the same time AeaeCec1-5 (1 μM each). And set up the control group, add an equal volume of PBS. After co-incubating for 6 hours, the cell culture supernatant was collected, and the cytokines tu...

Embodiment 3

[0064] Toxicity of Aedes aegypti Innate Immunity Polypeptide Cecropin to Mammalian Cells

[0065] C57BL / 6 mouse peritoneal macrophages, human PBMCs and Vero E6 cells of the green monkey kidney cell line were plated in 96-well culture plates (2×10 4 Cells / hole) were cultured with RMPI-1640 medium (100 μl / well, purchased from Gbico, USA) added with 2% fetal bovine serum, 100 U / ml ampicillin and 100 μg / ml streptomycin sulfate, until the cells adhered Afterwards, a series of peptides serially diluted 2-fold with serum-free RPMI-1640 medium were added, and after co-cultivation for 24 hours, cell proliferation and toxicity detection reagent CCK-8 (10 μl / well, purchased from Beijing Wobison Technology Co., Ltd. Company), after incubation for 4 hours, the light absorption at 450nm was detected, the cell viability without adding polypeptide was defined as 100%, and the cell viability after different concentrations of polypeptide treatment was calculated.

[0066] The results showed th...

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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 invention relates to the field of biomedicine, in particular to the application of a cecropin polypeptide as an anti-inflammatory drug. Background technique [0002] Infectious diseases have always been a global public problem that threatens human health. Patients infected by pathogenic microorganisms are usually accompanied by an uncontrollable inflammatory response of the body. For example, during bacterial infection, Gram-negative bacteria will release a large amount of cell wall components-lipopolysaccharide, and Gram-positive bacteria will also release a large amount of cell wall components-lipid muramic acid, while lipopolysaccharide and lipid Muramic acid is an agonist of Toll-like receptor 4 and Toll-like receptor 2, respectively, thereby activating a series of inflammatory signaling pathways downstream of immune cells, such as the mitogen-activated protein kinase signaling pathway MAPKs and the nuclear transcription factor signaling pathw...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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