Diclofenac and Meloxicam Exhibited Anti-Virulence Activities Targeting Staphyloxanthin Production in Methicillin-Resistant Staphylococcus aureus
Abstract
:1. Introduction
2. Results
2.1. Identification of Clinical Isolates
2.2. Antibiotic Resistance Pattern
2.3. Staphyloxanthin Production Assay
2.4. Quantitative Estimation of Staphyloxanthin Production
2.5. Determination of MICs of Tested Drugs
2.6. The Effect of the Tested Drugs on Staphyloxanthin Synthesis
2.7. Effect of Staphyloxanthin Inhibition on Protease Production
2.8. Effect of Staphyloxanthin Inhibition on DNase Activity
2.9. ERIC-PCR Analysis
2.10. Detection of mecA Gene among MRSA Isolates
2.11. DC/MXM Treatment Effect on Virulent Genes Expression
2.12. Molecular Docking
3. Discussion
4. Materials and Methods
4.1. Bacterial Strains
4.2. Stock Solutions
4.3. Antibiotic Susceptibility of Clinical Isolates
4.4. Staphyloxanthin Production Assay
4.5. Determination of MICs of Tested Drugs
4.6. Staphyloxanthin Inhibition Assay
4.7. Extracellular Protease Assay
4.8. Extracellular Nuclease Assay
4.9. Determination of Genetic Diversity Using ERIC-PCR
4.10. Detection of mecA Gene by cPCR
4.11. Real-Time PCR (qPCR) Analysis
4.12. Molecular Docking Analysis
4.13. Statistical Analysis
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Antimicrobial Agents | Susceptibility Pattern of the Clinical Isolates, (N = 80) | |||||
---|---|---|---|---|---|---|
Sensitive (S) | Intermediate (I) | Resistant (R) | ||||
N | % | N | % | N | % | |
Cefoxitin | 0 | 0 | 0 | 0 | 80 | 100 |
Clindamycin | 50 | 62.5 | 1 | 1.25 | 29 | 36.25 |
Ciprofloxacin | 44 | 55 | 3 | 3.75 | 33 | 41.25 |
Linezolid | 80 | 100 | 0 | 0 | 0 | 0 |
Vancomycin | 80 | 100 | 0 | 0 | 0 | 0 |
STX Inhibitors | cDOCKER Energy (Kcal/mol) | cDOCKER Interaction Energy (Kcal/mol) | Interactions with Amino Acid Residues of CrtM | |
---|---|---|---|---|
Diclofenac | −28.5335 | −36.6979 | Hydrogen bonds | Hydrophobic interactions |
2 H-bonds with ARG45 | 8 hydrophobic bonds with PHE22, TYR41, ALA134, VAL137 and LEU164 | |||
Meloxicam | −23.4162 | −40.4063 | 6 H-bonds with TYR41, ARG45, GLN165 and ASN168 | 2 hydrophobic bonds with ALA134 and VAL137 |
Target Gene | Primer Sequence (5′–3′) |
---|---|
crtM | F-CTGCTAATTCTATGATTGGTTGTGC |
R-TGGGAATATTATGCAGCTATMGCAG | |
crtN | F-GATGAAGCTTTGACGCAACA |
R-TTCGCATGATACGTTTGCTC | |
hla | F-GAA AGG TAC CAT TGC TGG TCA |
R-AAG GCC AGG CTA AAC CAC TT | |
sigB | F-AAG TGA TTC GTA AGG ACG TCT |
R-TCG ATA ACT ATA ACC AAA GCC T |
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Elmesseri, R.A.; Saleh, S.E.; Ghobish, S.A.; Majrashi, T.A.; Elsherif, H.M.; Aboshanab, K.M. Diclofenac and Meloxicam Exhibited Anti-Virulence Activities Targeting Staphyloxanthin Production in Methicillin-Resistant Staphylococcus aureus. Antibiotics 2023, 12, 277. https://doi.org/10.3390/antibiotics12020277
Elmesseri RA, Saleh SE, Ghobish SA, Majrashi TA, Elsherif HM, Aboshanab KM. Diclofenac and Meloxicam Exhibited Anti-Virulence Activities Targeting Staphyloxanthin Production in Methicillin-Resistant Staphylococcus aureus. Antibiotics. 2023; 12(2):277. https://doi.org/10.3390/antibiotics12020277
Chicago/Turabian StyleElmesseri, Rana A., Sarra E. Saleh, Sarah A. Ghobish, Taghreed A. Majrashi, Heba M. Elsherif, and Khaled M. Aboshanab. 2023. "Diclofenac and Meloxicam Exhibited Anti-Virulence Activities Targeting Staphyloxanthin Production in Methicillin-Resistant Staphylococcus aureus" Antibiotics 12, no. 2: 277. https://doi.org/10.3390/antibiotics12020277