The antibacterial mechanism of oridonin against methicillin-resistant Staphylococcus aureus (MRSA)

Pharm Biol. 2019 Dec;57(1):710-716. doi: 10.1080/13880209.2019.1674342.

Abstract

Context: Methicillin-resistant Staphylococcus aureus (MRSA) is a very harmful bacterium. Oridonin, a component in Rabdosia rubescens (Hemsl.) Hara (Lamiaceae), is widely used against bacterial infections in China. Objective: We evaluated oridonin effects on MRSA cell membrane and wall, protein metabolism, lactate dehydrogenase (LDH), DNA and microscopic structure. Materials and methods: Broth microdilution and flat colony counting methods were used to measure oridonin minimal inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) against USA300 strain. Electrical conductivity and DNA exosmosis were analysed to study oridonin effects (128 μg/mL) on cell membrane and wall for 0, 1, 2, 4 and 6 h. Sodium dodecyl sulphate polyacrylamide gel electrophoresis was used to detect effects on soluble protein synthesis after 6, 10 and 16 h. LDH activity was examined with an enzyme-linked immunosorbent assay. Effects of oridonin on USA300 DNA were investigated with DAPI staining. Morphological changes in MRSA following oridonin treatment were determined with scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Results: Oridonin MIC and MBC values against USA300 were 64 and 512 μg/mL, respectively. The conductivity and DNA exosmosis level of oridonin-treated USA300 improved by 3.20±0.84% and increased by 58.63 ± 1.78 μg/mL, respectively. LDH and soluble protein levels decreased by 30.85±7.69% and 27.51 ± 1.39%, respectively. A decrease in fluorescence intensity was reported with time. Oridonin affected the morphology of USA300. Conclusions: Oridonin antibacterial mechanism was related to changes in cell membrane and cell wall permeability, disturbance in protein and DNA metabolism, and influence on bacterial morphology. Thus, oridonin may help in treating MRSA infection.

Keywords: Bacteriostat; DNA; LDH; cell membrane; cell wall; microscopic structure; protein metabolism.

MeSH terms

  • Anti-Bacterial Agents / administration & dosage
  • Anti-Bacterial Agents / pharmacology*
  • Cell Membrane / drug effects*
  • Cell Wall / drug effects
  • DNA, Bacterial / metabolism
  • Diterpenes, Kaurane / administration & dosage
  • Diterpenes, Kaurane / pharmacology*
  • Methicillin-Resistant Staphylococcus aureus / drug effects*
  • Microbial Sensitivity Tests
  • Microscopy, Electron, Scanning
  • Microscopy, Electron, Transmission
  • Time Factors

Substances

  • Anti-Bacterial Agents
  • DNA, Bacterial
  • Diterpenes, Kaurane
  • oridonin

Grants and funding

The work was supported by National Natural Science Foundation of China (No. 31702284) and Double Subject Construction Plan of Sichuan Agriculture University (No. 03571444).