Application of deoxyshikonin in preparation of host defense peptide expression promoter

A technology of deoxyshikonin and host defense peptide, applied in the direction of application, additional food elements, medical preparations containing active ingredients, etc., to achieve the effect of improving antibacterial activity and immune function

Active Publication Date: 2021-02-26
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are few candidates with HDPs-induced expression, so it is urgent to study and find more compounds with HDPs-induced expression

Method used

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  • Application of deoxyshikonin in preparation of host defense peptide expression promoter
  • Application of deoxyshikonin in preparation of host defense peptide expression promoter
  • Application of deoxyshikonin in preparation of host defense peptide expression promoter

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] The screening of embodiment 1.Deoxyshikonin obtains

[0032] 1261 known pure natural products and their derivatives isolated from plants, animals, microorganisms, etc. belonging to more than 30 substances such as alkaloids, lipids, sequence terpenes, diterpenes, pentacyclic triterpenes, and sterols were tested. filter.

[0033] Obtained 48 kinds of compounds with inducing activity to pBD3 gene in IPEC-J2 / pBD3-luc stable reporter cells; through dose-response experiments, it was found that 21 kinds of compounds, including Deoxyshikonin, were tested at three concentrations (5μM, 20μM, 80μM) at least one concentration can increase luciferase activity more than 2-fold.

[0034] Experimental reagents:

[0035] IPEC-J2 / pBD3-luc luciferase reporter cells containing the porcine pBD3 gene promoter (recorded in the article published by the inventors: Deng, Z., et al. 2018. Development of a Cell-Based High-Throughput Screening Assay to Identify Porcine Host Defense Peptide-Induc...

Embodiment 2

[0047] Example 2. Verification of the inducible expression effect of Deoxyshikonin on pBD3 gene on IPEC-J2 cells

[0048] Experimental reagents:

[0049] Original cell line IPEC-J2: healthy piglet small intestinal epithelial cells, donated by Guolong Zhang, Oklahoma State University, USA, and can also be obtained through other commercial channels.

[0050] Deoxyshikonin stock solution: the concentration is 20 mM, the solvent is dimethyl sulfoxide (DMSO), purchased from TargetMol, Boston, MA, USA.

[0051] Experimental steps:

[0052] 1. Culture of IPEC-J2 cells

[0053] IPEC-J2 cell culture adopts DMEM / F12 medium, and its complete culture solution is composed of: DMEM / F12 medium+10% fetal bovine serum (FBS)+1% double antibody (penicillin / streptomycin).

[0054] The frozen cells were taken out of the liquid nitrogen tank and quickly put into a 37°C water bath to thaw. In a biological safety cabinet, suck out the cell suspension and transfer it completely to a centrifuge tub...

Embodiment 3

[0075] Example 3. Effect of Deoxyshikonin on the Induced Expression of Other Host Defense Peptides

[0076] The cultivation and stimulation of cells, the extraction of total RNA, and the determination of mRNA expression are the same as in Example 2, and the primer sequences used are as follows:

[0077] GAPDH, F: 5'-GCTACACTGAGGACCAGGTTG-3' (SEQ ID No. 1),

[0078] R: 5'-CCTGTTGCTGTAGCCAAATT-3'C (SEQ ID No.2);

[0079] pBD2, F: 5'-TGTCTGCCTCCTCTCTTCC-3' (SEQ ID No. 5),

[0080] R: 5'-AACAGGTCCCTTCAATCCTG-3' (SEQ ID No. 6);

[0081] PG1-5, F: 5'-ACGGTGAAGGAGACTGTG-3' (SEQ ID No. 7),

[0082] R: 5'-CGCAGAACCTACGCCTACAA-3' (SEQ ID No.8);

[0083] pEP2C, F: 5'-ACTGCTTGTTTCCAGAGCC-3' (SEQ ID No. 9),

[0084] R: 5'-TGGCACAGATGACAAAGCCT-3' (SEQ ID No. 10).

[0085] The test results are as follows: the compound Deoxyshikonin not only induces the expression of the host defense peptide pBD3 gene in pig IPEC-J2 cells, but also significantly induces the expression of the host defens...

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PUM

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Abstract

The invention discloses application of deoxyshikonin in preparation of a host defense peptide expression promoter, belongs to new application of a compound, and is characterized in that the deoxyshikonin or a derivative thereof is used as a pharmacodynamic active ingredient to be prepared into a required dosage form; wherein the promoter is used for promoting up-regulation expression of host defense peptide genes; and the compound has a good promotion effect on expression of various host defense peptides, does not cause inflammatory reaction, has the capabilities of regulating host endogenousimmunity and improving the body health level, and can be used as a candidate of a new generation of antibiotic substitutes.

Description

technical field [0001] The invention relates to the new application technology of the compound, in particular the application of deoxyshikonin in the preparation of host defense peptide expression promoter. Background technique [0002] Host defense peptides (HDPs) are important effector molecules of the body's innate immune system and an important barrier for the body to resist the invasion of foreign pathogenic bacteria. So far, there are as many as 2385 HDPs derived from animals (http: / / aps.unmc.edu / AP / main.php). Host defense peptides are small positively charged amphipathic peptides that can directly kill bacteria, fungi and viruses, or inhibit their growth. Host defense peptides also have powerful immune regulation functions, including stimulating immune cell proliferation, modifying related gene expression, promoting wound healing, killing cancer cells, etc. [0003] Inducing the synthesis of endogenous HDPs in the host is beneficial to the body's resistance to early...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/122A61P31/00A23K50/60A23K20/111A23K50/30A01K67/02
CPCA61K31/122A61P31/00A23K50/60A23K20/111A23K50/30A01K67/02
Inventor 王晶季海峰张伟王四新王雅民
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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