Expression of Thanatin in HEK293 Cells and Investigation of its Antibacterial Effects on Some Human Pathogens
| Autor: | Zahra Mousavi, Ali Javadmanesh, Abbas Tanhaeian, Marjan Azghandi |
|---|---|
| Rok vydání: | 2019 |
| Předmět: |
Calcium Phosphates
0301 basic medicine Cell Survival medicine.drug_class Antimicrobial peptides Antibiotics Gene Expression Peptide Microbial Sensitivity Tests Transfection Biochemistry Article Microbiology Antibiotic resistant Mice 03 medical and health sciences Minimum inhibitory concentration Structural Biology medicine Animals Humans MTT assay chemistry.chemical_classification Recombinant Bacteria Base Sequence 030102 biochemistry & molecular biology biology Chemistry General Medicine biology.organism_classification Antimicrobial Recombinant Proteins Anti-Bacterial Agents HEK293 Cells 030104 developmental biology NIH 3T3 Cells Pathogens Antibacterial activity Thanatin Antimicrobial Cationic Peptides |
| Zdroj: | Protein and Peptide Letters |
| ISSN: | 0929-8665 |
| DOI: | 10.2174/0929866526666190822162140 |
| Popis: | Background:Thanatin is the smallest member of Beta-hairpin class of cationic peptide derived from insects with vast activities against various pathogens.Objective:n this study, the antimicrobial activity of this peptide against some species of human bacterial pathogens as well as its toxicity on NIH cells were evaluated.Method:Thanatin DNA sequence was cloned into pcDNA3.1+ vector and transformed into a DH5α bacterial strain. Then the recombinant plasmids were transfected into HEK-293 cells by calcium phosphate co-precipitation. After applying antibiotic treatment, the supernatant medium containing thanatin was collected. The peptide quantity was estimated by SDS-PAGE and GelQuant software. The antimicrobial activity of this peptide was performed with Minimum Inhibitory Concentration (MIC) method. In addition, its toxicity on NIH cells were evaluated by MTT assay.Results:The peptide quantity was estimated approximately 164.21 µmolL-1. The antibacterial activity of thanatin was estimated between 0.99 and 31.58 µmolL-1 using MIC method. The result of cytotoxicity test on NIH cell line showed that the peptide toxicity up to the concentration of 394.10 µmolL-1 and for 48 hours, was not statistically significant from negative control cells (P>0.05). The antimicrobial assay demonstrated that thanatin had an antibacterial effect on some tested microorganisms. The results obtained in this study also showed that thanatin had no toxicity on mammalian cell lines including HEK293 and NIH.Conclusion:Antimicrobial peptides such as thanatin are considered to be appropriate alternatives to conventional antibiotics in treating various human pathological diseases bacteria. |
| Databáze: | OpenAIRE |
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