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Effect of apoptosis in neural stem cells treated with sevoflurane
Jianlei Qiu1, Pengcai Shi2, Wude Mao3
1Department of Anesthesiology, Dezhou People's Hospital, Dezhou, Shandong China ; School of Medicine, Shandong University, Ji'nan, Shandong China.
Insights
Sevoflurane anesthesia can harm infant brain development by increasing neural stem cell apoptosis via GABAA receptor activation. Blocking this receptor with bicuculline reversed these damaging effects, suggesting a potential protective strategy.
Area of Science:
- Neuroscience
- Anesthesiology
- Developmental Biology
Background:
- Infant exposure to sevoflurane anesthesia poses risks to developing brains.
- Long-term anesthetic exposure may cause neurological disorders and cognitive deficits.
- γ-Aminobutyric acid (GABA) is an inhibitory neurotransmitter implicated in anesthetic effects.
Purpose of the Study:
- To investigate the impact of sevoflurane on neural stem cells (NSCs).
- To determine the role of GABAA receptors in sevoflurane-induced neurotoxicity.
- To evaluate the neuroprotective potential of GABAA receptor antagonists.
Main Methods:
- Primary neural stem cells were cultured and exposed to sevoflurane with or without bicuculline, a GABAA receptor antagonist.
- Cell viability (CCK-8), cytotoxicity (LDH), and apoptosis (Annexin V/PI staining) were assessed.
- Expression levels of GABAA receptor subunits, Bcl-2, Bax, and Caspase-3 were analyzed via Western blotting.
Main Results:
- Sevoflurane exposure dose-dependently decreased cell viability and increased cytotoxicity and apoptosis.
- Sevoflurane upregulated GABAA receptor, Bax, and Caspase-3, while downregulating Bcl-2.
- Bicuculline treatment effectively reversed the detrimental effects of sevoflurane on NSCs.
Conclusions:
- Sevoflurane induces neural stem cell apoptosis and degeneration by activating GABAA receptors.
- GABAA receptor antagonism mitigates sevoflurane-induced neurotoxicity in developing neural stem cells.
Background:
At present, sevoflurane inhalation anesthesia used on infants is well-known. But long-time exposure to inhalation anesthetic could cause neurologic disorder, especially nerve degeneration in infant and developing brain. The central nervous system degeneration of infants could affect the memory and cognitive function. γ-Aminobutyric acid (GABA) is a known inhibitory neurotransmitter in central nervous system. Inhalation anesthetic sevoflurane may activate GABAA receptor to inhibit central nervous system, leading to apoptosis of neural degeneration, cognitive dysfunction in the critical period of brain development.
Methods:
Neural stem cells were derived from Wistar embryos, cultured in vitro. Third generation of neural stem cells were randomly divided into four groups according to cultured suspension: Sevoflurane group (Group S), GABAA receptor antagonists, Bicuculline group (Group B), Sevoflurane + GABAA receptor antagonists, Bicuculline group (Group S + B), dimethyl sulphoxide (DMSO) group (Group D). Group B and Group D did not receive sevoflurane preconditioning. Group S and Group S + B were pretreated with 1 minimum alveolar concentration (MAC) sevoflurane for 0 h, 3 h, 6 h, and 12 h. Group S + B and Group B were pretreated with bicuculline (10 uM). Group D was treated with DMSO (10 uL/mL). After treatments above, all groups were cultured for 48 h. Then we measured the cells viability by Cell Counting Kit (CCK-8) assay, cytotoxicity by Lactate Dehydrogenase (LDH) assay, apoptosis ratio with Annexin V/propidium iodide (PI) staining by flow cytometry, and the expression of GABAAR, anti-apoptotic protein Bcl-2, pro-apoptotic protein Bax and Caspase-3 by western blotting.
Results:
After exposing to sevoflurane for 0 h, 3 h, 6 h, and 12 h with 1MAC, we found that cell viability obviously decreased and cytotoxicity increased in time-dependent way. And Annexin V/PI staining indicated increased apoptosis ratio by flow cytometry. The protein level of GABAA receptor, pro-apoptotic protein Bax and apoptosis protein Caspase-3 increased; while anti-apoptotic protein Bcl-2 decreased. And bicuculline could reverse all detrimental results caused by sevoflurane.
Conclusion:
Sevoflurane can inhibit the central nervous system by activating GABAA, resulting in apoptosis of neural stem cells, thus leading to the NSCs degeneration.
