Role of PARP1 on DNA damage induced by mineral silicate chrysotile in bronchial epithelial and pleural mesothelial cells.

To investigate whether poly (ADP ribose) polymerase-1 (PARP1) is involved in chrysotile-induced DNA damage in pleural mesothelial cells (MeT-5A) and bronchial epithelial cells (BEAS-2B), two PARP1-deficient cell lines were established. Efficiencies of RNA interference on PARP1 were detected by western blot and qPCR. Here, normal cells and PARP1-deficient cells were exposed to chrysotile, and DNA damage and DNA repair were detected by alkaline comet assay. All cells were treated with chrysotile at the indicated concentrations (5, 10, 20, and 40 µg/cm2) for 24 h and then the DNA repair capacity was observed for 12 and 24 h, respectively. The results showed that chrysotile caused DNA damage at an obvious dose-dependent manner in MeT-5A and BEAS-2B cells. In addition, MeT-5A cells had more persistent DNA damage than BEAS-2B. Compared to normal cells, the PARP1-deficient cells were more sensitive to DNA damage caused by chrysotile. In DNA repair experiments, all cell lines recovered from the damage over time. The results of relative repair percentage (RRP) of MeT-5A and BEAS-2B were higher than those of MeT-5A shPARP1 and BEAS-2B shPARP1 cells at all experimental concentrations (except 5 µg/cm2) at 12-h repair. However, RRP of BEAS-2B and BEAS-2B shPARP1 tended to be closer, and RRP of MeT-5A shPARP1 was still lower than that of MeT-5A at 24-h repair. All results suggest that PARP1 plays an important role in early repair of DNA damage in BEAS-2B and MeT-5A cells exposed to chrysotile.

Azalomycin F5a, a polyhydroxy macrolide binding to the polar head of phospholipid and targeting to lipoteichoic acid to kill methicillin-resistant Staphylococcus aureus.

Azalomycin F5a was a polyhydroxy macrolide produced by streptomycete strains. Our preliminary researches indicated that it could kill methicillin-resistant Staphylococcus aureus (MRSA) likely by increasing the permeability of cell membrane, and that cell-membrane phospholipids were likely important targets. To confirm this, membrane permeability assay was performed and visualized by fluorescence staining, and then the detailed interactions between azalomycin F5a and model membranes prepared with 1,2-dihexadecanoyl-sn-glycero-3-phospho-(1'-rac-glycerol) (DPPG) were determined using attenuated total reflectance fourier transform infrared spectroscopy and 31P nuclear magnetic resonance techniques. The results indicated that there were strong interactions between azalomycin F5a and model membranes, especially between azalomycin F5a and the polar head of phospholipid. For further evidence and details, the molecular dynamics (MD) simulation of the interactions between azalomycin F5a and DPPG or lysyl-DPPG were performed using Amber16 software package. A strong interaction between the lactone ring of azalomycin F5a and the polar head of DPPG or lysyl-DPPG had been clearly observed. Moreover, a larger distribution probability out of phospholipid bilayer had been discovered for the guanidyl side chain of azalomycin F5a, especially when probable anion molecules anchoring on the cytoplasmic membrane occurred. Therefore, lipoteichoic acid (LTA), a vital component of gram-positive bacterial envelope, was investigated for its probable interactions with azalomycin F5a using broth microdilution method. The results showed that azalomycin F5a-induced MRSA lysis could be prevented by LTA. This deduced that there were some interactions between azalomycin F5a, more likely its guanidyl side chain, and LTA. Thereby, azalomycin F5a increasing the cell-membrane permeability of MRSA had likely achieved by the synergy of its lactone ring binding to the polar head of phospholipid and its guanidyl side chain targeting to LTA, and which had eventually led to the autolysis of MRSA cells.