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Investigating the Effects of Osmolytes and Environmental pH on Bacterial Persisters.

ABSTRACT: Bacterial persisters are phenotypic variants that temporarily demonstrate an extraordinary tolerance toward antibiotics. Persisters have been linked to the recalcitrance of biofilm-related infections; hence, a complete understanding of their physiology can lead to improvement of therapeutic strategies for such infections. Mechanisms pertaining to persister formation are thought to be associated with stress response pathways triggered by intra- or extracellular stress factors. Unfortunately, studies demonstrating the effects of osmolyte- and/or pH-induced stresses on bacterial persistence are largely missing. To fill this knowledge gap within the field, we studied the effects of various osmolytes and pH conditions on Escherichia coli persistence with the use of phenotype microarrays and antibiotic tolerance assays. Although we found that a number of chemicals and pH environments, including urea, sodium nitrite, and acidic pH, significantly reduced persister formation in E. coli compared to no-osmolyte/no-buffer controls, this reduction in persister levels was less pronounced in late-stationary-phase cultures. Our results further demonstrated a positive correlation between cell growth and persister formation, which challenges the general notion in the field that slow-growing cultures have more persister cells than fast-growing cultures.

SUBMITTER: Karki P

PROVIDER: S-EPMC7179588 | biostudies-literature | 2020 Apr

REPOSITORIES: biostudies-literature

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Investigating the Effects of Osmolytes and Environmental pH on Bacterial Persisters.

Karki Prashant P Mohiuddin Sayed Golam SG Kavousi Pouria P Orman Mehmet A MA

Antimicrobial agents and chemotherapy 20200421 5

Bacterial persisters are phenotypic variants that temporarily demonstrate an extraordinary tolerance toward antibiotics. Persisters have been linked to the recalcitrance of biofilm-related infections; hence, a complete understanding of their physiology can lead to improvement of therapeutic strategies for such infections. Mechanisms pertaining to persister formation are thought to be associated with stress response pathways triggered by intra- or extracellular stress factors. Unfortunately, stud ...[more]

PMID: 32094133

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Project description:Background:Proton pump inhibitor (PPI) and other acid-suppressing drugs are widely used in the treatment of gastrointestinal ulcer, upper gastrointestinal bleeding, gastritis, and gastric cancer (GC). About 80% of GC patients receive acid suppression treatment. PPI suppresses the production of gastric acid by inhibiting the function of H+/K+-ATPase in gastric parietal cells and raises the pH value to achieve therapeutic purposes. Some studies have found that PPI had a certain antitumor effect in the proliferation and apoptosis of tumor cells. But the effects of environmental pH on the growth of GC cells and its mechanism are unknown. Therefore, we hoped to find the effects of culture medium pH on the biological behavior of GC cells by in vitro experiments and provide guidance for the use of acid-suppressing drugs in GC patients. Aims:We aimed to observe the effects of pH changes in GC cell culture medium on the cell biological behavior of cancer cells and to analyze the potential mechanisms. We hoped to find out the effect of acid suppression on the growth of GC cells. Methods:The GC cell lines (SGC-7901 and MKN45) were used as the research object. We adjusted the pH value in the cell culture medium to observe the changes in cell viability (MTT), apoptosis (flow cytometry), and invasion (Transwell) at pH?6, pH?7, and pH?8. qRT-PCR and western blot (WB) assays were used to determine the expression changes of genes and proteins (mTOR, AKT, Wnt, Glut, and HIF-1?) at pH?6, pH?7, and pH?8. Results:The results of MTT showed that the viability of SGC-7901 and MKN45 in the pH?8.0 group was significantly weaker than that in the pH?6.0 or pH?7.0 group (P < 0.001). Flow cytometry results showed that the apoptosis of SGC-7901 and MKN45 in the pH?8.0 group was more obvious than that in the pH?6.0 or pH?7.0 group (P < 0.001). Flow cytometry results showed that the apoptosis of SGC-7901 and MKN45 in the pH?8.0 group was more obvious than that in the pH?6.0 or pH?7.0 group (P < 0.001). Flow cytometry results showed that the apoptosis of SGC-7901 and MKN45 in the pH?8.0 group was more obvious than that in the pH?6.0 or pH?7.0 group (?) at pH?6, pH?7, and pH?8. P < 0.001). Flow cytometry results showed that the apoptosis of SGC-7901 and MKN45 in the pH?8.0 group was more obvious than that in the pH?6.0 or pH?7.0 group (. Conclusions:Compared with the microacid environment, the microalkaline environment inhibited the viability, invasion, and expression of genes and proteins (mTOR, AKT, Wnt, Glut, and HIF-1?) but promoted the apoptosis of GC cells and thus inhibited the growth of GC.?) at pH?6, pH?7, and pH?8.

Dataset Information

Investigating the Effects of Osmolytes and Environmental pH on Bacterial Persisters.

Publications

Investigating the Effects of Osmolytes and Environmental pH on Bacterial Persisters.

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