Abstract
Purpose: Neural injury affects patients after using inhalational anesthetics such as sevoflurane. Rhodioloside, a compound which is obtained from the Rhodiola rosea plant has been implicated to be the most commonly used psychostimulant that can improve a range of conditions. The study was aimed at finding the molecular mechanism underlying the Rhodioloside treatment of sevoflurane-injured hippocampal neurons.Methods: Main hippocampal neurons, secluded from Sprague Dawley embryonic rats were employed to create an injury model using 3 % sevoflurane. The sevoflurane-injured hippocampal neurons were treated with varying concentrations (10, 20, 40 and 80 μM/ml) of Rhodioloside to create different experimental groups: RHSD10+SEV, RHSD20+SEV, RHSD40+SEV, RHSD80+SEV, while untreated cells were considered as the Control group. Cell viability was identified using the CCK-8 assay. The CFSE assay was used to verify the promotion function of Rhodioloside on cell differentiation of neurons. FCM assay was employed to determine cell proliferation and apoptosis. Expression levels of apoptosisrelated factors, like Caspase-3, Bcl-2 and Bax were examined by RT-qPCR, while Western blot was used to measure phosphorylation of PKA.Results: Rhodioloside stimulated cell viability and prevented cell apoptosis in sevoflurane-injured hippocampal neurons in doses between 10-80 μM. The apoptosis-inhibitory effect of Rhodioloside was observed to be through cAMP/PKA pathway activation. Also, expression levels of Bcl-2, and PKA were enhanced and the level of Caspase-3 and Bax was reduced in a dose-dependent pattern. The PKA inhibitor reversed the above observation in the 40 μM Rhodioloside-treatment.Conclusion: Rhodioloside promoted cell viability and prevented apoptosis of primary hippocampal neurons injured by sevoflurane, through cAMP/PKA pathway activation. Inhibition of PKA network deteriorated the function of Rhodioloside by stimulating cell apoptosis. Our findings present a novel evidence that Rhodioloside could attenuate neurotoxicity of inhalational anesthetics.
 Keywords: Cell apoptosis, cAMP/PKA pathway, Hippocampal neurons, Rhodioloside, Sevoflurane
Highlights
Sevoflurane is among the most used anesthetics in cesarean delivery and pediatric clinical process due to its swift induction and recovery features
CCK-8 assay was used to detect the function of rhodioloside with different concentrations (10, 20, 40 and 80 μM/ml) on cell viability of hippocampal primary neurons treated with 3 % sevoflurane, at 6 hr
It would be better to examine the effect of protein kinase (PKA) activation on it at the same time, the results have indicated that rhodioloside protected neurons from sevoflurane induced apoptosis through cAMP/PKA pathway, which might reduce cell viability and proliferative ability of primary hippocampal neurons
Summary
Sevoflurane is among the most used anesthetics in cesarean delivery and pediatric clinical process due to its swift induction and recovery features. Studies have shown over the years that the use of anesthesia in youngsters below 3 years could impact their development and behavior, thereby leading to cognitive disorders [1]. It is abundantly and widely distributed in China. In Europe and Asia, R. rosea is mentioned as the most commonly used psychostimulant that can improve a range of conditions, including depression, anxiety, fatigue, anaemia, endurance, altitude sickness, insomnia and impotence [10,11]. A recent study found that the plant’s supplement strikingly improves aerobic exercise performance after short-term high altitude training and has lots of biological activities, such as anti-stress and anti-fatigue [12,13]. Accumulating evidence indicates that the plant extracts protect against cerebral ischemia-reperfusion injury in rat brain [15]
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