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Neuroprotective effect of estradiol and progesterone against hypoxia-induced neuronal damage in vitro
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Zeinab Vahidinia   , Abolfazl Azami Tameh , Javad Amini Mahabadi |
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Abstract: (321 Views) |
Introduction: Hypoxia induces neuronal injury and previous reports have indicated that 17β-estradiol (E2) and progesterone (P) have neuroprotective effects in neurological disorders and in particular under hypoxic conditions; however, little is known in regards to their pharmacological mechanism. In this study, we tried to evaluate the neuroprotective effects and mechanisms of E2, P, and a combination of both hormones against hypoxia injury using an in vitro model.
Methods and Materials: The hypoxia injury model was produced by hypoxia/reoxygenation (H/R). Primary cortical neurons were obtained from cerebral cortices of 12-day-old mouse embryos and cultured in vitro. Cortical neurons were exposed to hypoxia for 24 h and treated with E2, P and EP in same condition. Microtubule-associated protein 2 (MAP2) immunostaining was performed to determine cortical neurons. Cell viability was detected by MTS assay and cell damage was validated by LDH release assay. Propidium iodide (PI)/hoechst (HO) was used to assess apoptosis rate of cortical neurons under H/R conditions. Western blot assay was carried out to measure the protein expression of cleaved caspase 3
Results: MAP2 immunostaining showed that most of the cultured cells were neurons. Exposure to hypoxia significantly reduced cell viability and increased neurons death. Moreover, expression levels of cleaved caspase3 was elevated in hypoxia induced groups. Hormone therapy significantly increased cell viability and reduced neurons death under hypoxia. Moreover, to elucidate a possible mechanism by which E2 and P exert their neuroprotective effect, we investigated the cleaved caspase3 using western blot assay. However, no significant differences were observed in the expression of cleaved caspase-3 between different groups.
Conclusion: Overall, we demonstrated that E2 or P or combined EP treatment alleviate hypoxia injury in vitro. However, hormone therapy had no effect on caspase activity. This study deepens our understanding of the mechanisms underlying hormone therapy-mediated neuroprotection.
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| Keywords: Hypoxia, 17β-estradiol, progesterone, Caspase 3, Cell apoptosis |
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Full-Text [PDF 333 kb]
(55 Downloads)
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Type of Study: Research |
Subject:
General
Received: 2024/02/3 | Accepted: 2023/08/23 | Published: 2023/08/23
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