International Journal of Peptide Research and Therapeutics, ( ISI ), Volume (19), No (4), Year (2013-7) , Pages (373-380)

Title : ( The Effect of Antimicrobial Peptide Temporin-Ra on Cell Viability and Gene Expression of Pro-inflammatory Factors in A549 Cell Line )

Authors: fateme asadi , Ahmad Asoodeh , reyhaneh kashef , Mohammad Reza Housaindokht , Alireza Haghparast , Jamshidkhan Chamani ,

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Antimicrobial peptide Temporin-Ra was isolated from the skin secretions of marsh frog Rana ridibunda. Temporin-Ra was chemically synthesized and purified using RP-HPLC technique. The cytotoxicity of peptide on lung airway epithelial cell line (A549) and peripheral blood mononuclear cells (PBMC) was studied by MTT assay.Furthermore, the effect of Temporin-Ra on the expression of pro-inflammatory factors such as IL-1b and IL-8 in A549 cell line was evaluated at peptide concentrations of 15, 30 and 50 lg/mL for 6, 12 and 24 h using semi-quantitative RT-PCR and real-time PCR methods. The result of our experiments revealed that Temporin-Ra decreased cell viability about 3–13 % in A549 cells and 4–6 % in PBMC cells. Moreover, Temporin-Ra induced the mRNA expression of IL-1b and IL-8 genes in a dose- and time-dependent manner. According to our results, maximum IL-8 mRNA expression was observed after a 24-h treatment of cancer cells with 50 lg/mL peptide with approximately 18-fold increase in expression. The least expression level of IL-1b was observed after 6-h of incubation in the presence of 15 lg/mL peptide with 2.5-fold increase in expression whereas the most expression level was obtained following 24 h-treatment with 50 lg/mL peptide with 26-fold increase in mRNA expression. Eventually, the present study highlights the role of Temporin-Ra as a potent antimicrobial peptide in the activation and maintenance of nflammatory processes.

Keywords

, Temporin-Ra  Pro-inflammatory factors , A549 cancer cell line  HPLC  MTT assay  Real time,
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@article{paperid:1035713,
author = {Asadi, Fateme and Asoodeh, Ahmad and Kashef, Reyhaneh and Housaindokht, Mohammad Reza and Haghparast, Alireza and Jamshidkhan Chamani},
title = {The Effect of Antimicrobial Peptide Temporin-Ra on Cell Viability and Gene Expression of Pro-inflammatory Factors in A549 Cell Line},
journal = {International Journal of Peptide Research and Therapeutics},
year = {2013},
volume = {19},
number = {4},
month = {July},
issn = {1573-3149},
pages = {373--380},
numpages = {7},
keywords = {Temporin-Ra  Pro-inflammatory factors ; A549 cancer cell line  HPLC  MTT assay  Real time; PCR},
}

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%0 Journal Article
%T The Effect of Antimicrobial Peptide Temporin-Ra on Cell Viability and Gene Expression of Pro-inflammatory Factors in A549 Cell Line
%A Asadi, Fateme
%A Asoodeh, Ahmad
%A Kashef, Reyhaneh
%A Housaindokht, Mohammad Reza
%A Haghparast, Alireza
%A Jamshidkhan Chamani
%J International Journal of Peptide Research and Therapeutics
%@ 1573-3149
%D 2013

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