Bioactive Secondary Metabolites of a Marine Bacillus sp. Inhibit Superoxide Generation and Elastase Release in Human Neutrophils by Blocking Formyl Peptide Receptor 1
Abstract
:1. Introduction
2. Results and Discussion
2.1. IA Inhibits FMLP-Induced Superoxide Generation and Elastase Release in Human Neutrophils
2.2. IA Does not Show Inhibition in Cell-Free Systems
2.3. Protein Kinase A (PKA) Pathway Does not Mediate the Inhibitory Effects of IA
2.4. IA Attenuates Ca2+ Mobilization Induced by FMLP
2.5. IA Inhibits Mitogen-Activated Protein (MAP) Kinases and AKT Phosphorylation in FMLP-Activated Neutrophils
2.6. IA Inhibits the Binding of N-Formyl-Nle-Leu-Phe-Nle-Tyr-Lys-fluorescein (FNLFNYK) in Human Neutrophils
2.7. IA Inhibits the Binding of FNLFNYK in HEK293 Cells Transfected with FPR1
2.8. The High Performance Liquid Chromatography (HPLC) Method Used for Identification of the Composition of IA
3. Experimental
3.1. Reagents
3.2. Isolation of Human Neutrophils
3.3. Expression of FPR1 in Human Embryonic Kidney (HEK293) Cells
3.4. Bacterial Strains, Cultivation Condition, and Crude Extract Preparation
3.5. Measurement of Superoxide Generation and Elastase Release
3.6. Superoxide and DPPH Scavenging Assay
3.7. Evaluation of LDH Release
3.8. Receptor Binding Assay
3.9. Measurement of Intracellular Calcium Concentration ([Ca2+]i)
3.10. Immunoblotting Analysis
3.11. HPLC Fingerprint of IA
3.12. Statistics Analysis
4. Conclusions
Acknowledgments
Conflicts of Interest
References
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Yang, S.-C.; Lin, C.-F.; Chang, W.-Y.; Kuo, J.; Huang, Y.-T.; Chung, P.-J.; Hwang, T.-L. Bioactive Secondary Metabolites of a Marine Bacillus sp. Inhibit Superoxide Generation and Elastase Release in Human Neutrophils by Blocking Formyl Peptide Receptor 1. Molecules 2013, 18, 6455-6468. https://doi.org/10.3390/molecules18066455
Yang S-C, Lin C-F, Chang W-Y, Kuo J, Huang Y-T, Chung P-J, Hwang T-L. Bioactive Secondary Metabolites of a Marine Bacillus sp. Inhibit Superoxide Generation and Elastase Release in Human Neutrophils by Blocking Formyl Peptide Receptor 1. Molecules. 2013; 18(6):6455-6468. https://doi.org/10.3390/molecules18066455
Chicago/Turabian StyleYang, Shun-Chin, Chwan-Fwu Lin, Wen-Yi Chang, Jimmy Kuo, Yin-Ting Huang, Pei-Jen Chung, and Tsong-Long Hwang. 2013. "Bioactive Secondary Metabolites of a Marine Bacillus sp. Inhibit Superoxide Generation and Elastase Release in Human Neutrophils by Blocking Formyl Peptide Receptor 1" Molecules 18, no. 6: 6455-6468. https://doi.org/10.3390/molecules18066455