Anti-inflammatory activity of cecropin-A2 from Musca domestica
| Autor: | Xu Bin, Meng-Fei Zhao, Shaoyu Li, Yan-Yan Xi, Fengxian Wei, Wen-Jia Li, Bai Jie, Rui-Yang Wei |
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| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Staphylococcus aureus animal structures 030106 microbiology Anti-Inflammatory Agents Biology Real-Time Polymerase Chain Reaction medicine.disease_cause Microbiology Proinflammatory cytokine law.invention Mice 03 medical and health sciences law Houseflies medicine Animals Immunologic Factors Gene Expression Profiling Transfection Molecular biology RAW 264.7 Cells 030104 developmental biology Infectious Diseases Cecropin Cell culture Recombinant DNA Cytokines Insect Proteins Lipoteichoic acid Signal transduction Antimicrobial Cationic Peptides |
| Zdroj: | Microbial Pathogenesis. 110:637-644 |
| ISSN: | 0882-4010 |
| Popis: | This study aimed to investigate the anti-inflammatory activity of Musca domestica cecropin-A2 (Mdc-A2) toward Staphylococcus aureus (S. aureus) to learn more about their immunological functions. RAW264.7 cells were transfected with recombinant lentiviruses introduce pLEX-Mdc-A2into the RAW264.7 cell line (RAW-Mdc-A2). The RAW264.7 cell line with empty pLEX (RAW-pLEX) was produced in the same manner as a negative control. Real-time quantitative reverse transcription PCR (RT-PCR) was performed to analyze the mRNA expression of TNF-a, IL-1β, NFκB-1 and NFκB-2 in S. aureus-stimulated RAW-Mdc-A2 cells and RAW-pLEX cells in untreated cells and cells treated for 3 h, 6 h, 12 h and 24 h. RT-PCR was performed to analyze the mRNA expression of TNF-a, NFκB-1 and NFκB-2 stimulated by Lipoteichoic acid (LTA). Production of TNF-a was detected by enzyme-linked immunosorbent assay (ELISA). Colony counts were used to calculate the number of CFU per mL of cell culture supernatants. The results showed that compared to RAW-pLEX cells, stable transfection of Mdc-A2 in RAW264.7 cells stimulated by S. aureus significantly down-regulated the mRNA expression of TNF-a transcript variant 1 (TNF-a-tv-1) at 6 h and 12 h and the mRNA expression of TNF-a transcript variant 2 (TNF-a-tv-2) at 3 h, 6 h and 12 h. Compared to RAW-pLEX cells, stable transfection of Mdc-A2 in RAW264.7 cells stimulated by S. aureus significantly down-regulated the mRNA expression of IL-1β-T at 3 h, 6 h and 12 h as well as the mRNA expression of IL-1β at 3 h and 6 h. The expression and production of TNF-a and bacterial burden of cell culture supernatants were significantly down-regulated in RAW-Mdc-A2 cells stimulated by S. aureus, and the expression and production of TNF-a were significantly down-regulated in RAW-Mdc-A2 cells stimulated by LTA. Compared to RAW-pLEX cells, stable transfection of Mdc-A2 in RAW264.7 cells stimulated by S. aureus significantly down-regulated the mRNA expression of NFκB-1 at 3 h, 6 h and 12 h as well as the mRNA expression of NFκB-2 at 6 h. Additionally, stable transfection of Mdc-A2 in RAW264.7 cells stimulated by LTA significantly down-regulated the mRNA expression of NFκB-1. In conclusion, Mdc-A2 possesses potent anti-inflammatory activity and potent antimicrobial activity. Additionally, Mdc-A2 may interact with LTA and execute strong anti-inflammatory activity by blocking the activation of NF-κB signaling pathways. |
| Databáze: | OpenAIRE |
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