Gentamycin Rationally Repositioned to Inhibit miR-34a Ameliorates Oxidative Injury to PC12 Cells
Zhila Izadi1,2, Ebrahim Barzegari3, Amin Iranpanah1,2
1Pharmaceutical Sciences Research Center, Health Institute, Kermanshah University of Medical Sciences, Kermanshah 6715847141, Iran.
Abstract:
Ischemic stroke accompanies oxidative stress and cell death in the cerebral tissue. The microRNA miR-34a plays a pivotal role in this molecular pathology. This study presents the rational repositioning of aminoglycosidic antibiotics as miR-34a antagonists in order to assess their efficiency in protecting the PC12 stroke model cells from oxidative stress occurring under cerebral ischemic conditions. A library of 29 amino-sugar compounds were screened against anticipated structural models of miR-34a through molecular docking. MiR-ligand interactions were mechanistically studied by molecular dynamics simulations and free-energy calculations. Cultured PC12 cells were treated by H2O2 alone or in combination with gentamycin and neomycin as selected drugs. Cell viability and apoptosis were detected by 3-(4,5-dimethylthiazolyl-2)-2,5-diphenyltetrazolium bromide (MTT) and annexin V-FITC/propidium iodate (PI) double staining assays, respectively. The expression levels of key factors involved in cell proliferation, oxidative stress, and apoptosis in treated PC12 cells were measured through a quantitative real-time polymerase chain reaction and flow cytometric annexin V-FITC/PI double staining assays. A stable and energetically favorable binding was observed for miR-34a with gentamycin and neomycin. Gentamycin pretreatments followed by H2O2 oxidative injury led to increased cell viability and protected PC12 cells against H2O2-induced apoptotic events. This study will help in further understanding how the suppression of miR-34a in neural tissue affects the cell viability upon stroke.
Insights
Aminoglycosidic antibiotics, like gentamycin, can act as antagonists to microRNA miR-34a. This study shows they protect neural cells from oxidative stress and cell death, offering potential stroke treatments.
Area of Science:
- Neuroscience
- Molecular Biology
- Pharmacology
Background:
- Ischemic stroke causes oxidative stress and neuronal cell death.
- MicroRNA miR-34a is implicated in the molecular pathology of stroke.
- Developing neuroprotective strategies against stroke-induced damage is crucial.
Purpose of the Study:
- To investigate the potential of repositioning aminoglycosidic antibiotics as miR-34a antagonists.
- To evaluate the neuroprotective efficacy of these compounds against oxidative stress in a PC12 cell stroke model.
Main Methods:
- Screening of 29 amino-sugar compounds against miR-34a using molecular docking.
- Mechanistic study of miR-ligand interactions via molecular dynamics simulations and free-energy calculations.
- Assessing cell viability (MTT assay) and apoptosis (Annexin V-FITC/PI staining) in PC12 cells treated with H2O2 and antibiotics.
Main Results:
- Stable and favorable binding was observed between miR-34a and the antibiotics gentamycin and neomycin.
- Gentamycin pretreatment significantly increased cell viability and protected PC12 cells against H2O2-induced apoptosis.
- Key factors related to cell proliferation, oxidative stress, and apoptosis were modulated by the treatments.
Conclusions:
- Aminoglycosidic antibiotics can function as miR-34a antagonists.
- Gentamycin demonstrates neuroprotective effects against oxidative stress relevant to ischemic stroke.
- Suppression of miR-34a may be a viable therapeutic strategy for enhancing neural cell viability post-stroke.
