Sevoflurane‑induced neurotoxicity is driven by OXR1 post‑transcriptional downregulation involving hsa‑miR‑302e

Abstract

Sevoflurane is a common anesthetic agent used in surgical settings and previous studies have indicated that it exerts a neurotoxic effect. However, the molecular mechanism underlying this side effect is unknown. In addition, the human microRNA‑302 (hsa‑miR‑302) family members have been reported to be involved in neuronal cell development and biology. Thus, the present study aimed to investigate the potential implication of hsa‑miR‑302e in the sevoflurane‑induced cytotoxicity on human hippocampal cells (HN‑h). HN‑h cells were transfected with hsa‑miR‑302e mimic, hsa‑miR‑302e inhibitor or negative controls and subsequently exposed to different concentrations of sevoflurane. An MTT assay was used to assess the cytotoxicity of sevoflurane on HN‑h cells. Cell apoptosis was determined by flow cytometry. The levels of lactate dehydrogenase release, reactive oxygen species, lipid peroxidation and intracellular calcium (Ca2+) were additionally detected. Reverse transcription‑quantitative polymerase chain reaction and western blotting were conducted to determine mRNA and protein expression, respectively. A luciferase assay was performed for validating the targeting of OXR1 by hsa‑miR‑302e. The results indicated that sevoflurane induced a decrease in cell viability, malondialdehyde and reactive oxygen species production, lactate dehydrogenase release, intracellular Ca2+ production, calcium/calmodulin‑dependent protein kinase II phosphorylation and apoptosis. In addition, treatment with sevoflurane induced the expression of hsa‑miR‑302e while the expression of its target, oxidation resistance gene 1 (OXR1), was significantly downregulated. Inhibition of hsa‑miR‑302e expression protected neuronal cells from sevoflurane cytotoxicity. Mechanistic studies demonstrated that OXR1 was a direct target of hsa‑miR‑302e. Furthermore, the overexpression of OXR1 abolished the effect of sevoflurane on neuronal cells. The results of the present study indicated that sevoflurane exerts its neurotoxic effect by regulating the hsa‑miR‑302e/OXR1 axis. Therefore, the manipulation of the hsa‑miR‑302e/OXR1 pathway will be useful for preventing sevoflurane‑induced neurotoxicity.

6-Shogaol Inhibits Oxidative Stress-Induced Rat Vascular Smooth Muscle Cell Apoptosis by Regulating OXR1-p53 Axis

“…It participates in the detoxification of ROS and exhibits an important role in protecting various human cells against oxidative damage (Li et al, 2014;Svistunova et al, 2019). Different from previous reports that OXR1 expression was suppressed under stress (Ingram et al, 2007;Yang et al, 2018), OXR1 showed upregulation upon H 2 O 2 exposure in the present study. Then, pre-treatment with 6S effectively reversed the upregulation of OXR1 in a dose-dependent manner (Figure 2), indicating that OXR1 might play important roles in the protective effects of 6S against apoptosis.…”

Preventing Neurodegeneration by Controlling Oxidative Stress: The Role of OXR1

“…Sevoflurane treatment increases levels of oxidative damage and cell death. However, experimentally increasing OXR1 levels abolished the ROS-associated toxicity of sevoflurane (Yang et al, 2018). Piperlongumine functions by a completely different mechanism.…”

miR-424 inhibits apoptosis and inflammatory responses induced by sevoflurane through TLR4/MyD88/NF-κB pathway

“…Primer sequences were as follows: U6 forward primer (FP), 5′-GTGCTCGCTTCGGCAGCACATATAC-3′, reverse primer (RP), 5′-AAAAATATGGAACGCTTCACGAATTTG-3; GAPDH FP, 5′-ACCCACTCCTCCACCTTTGA-3′, RP, 5′-CTGTTGCTGTAGCCAAATTCGT-3′; hsa-miR-424 FP, 5′-CGCAAAACGTGAGGCGCT-3′ -3′, RP, 5-CCAGTGCAGGGTCCGAGGTA-3′; TLR4 FP, 5′-AATGGATCAAGGACCAGAGG-3′, RP, 5′-CAGCCAGCAAGAAGCATCAG-3′. The PCR program consisted of 30 s at 95 °C, followed by 45 cycles of 5 s at 95 °C and 58 °C for 34 s. The expression level of miRNA was obtained using the 2 − ∆∆Cq method with U6 small nuclear RNA as control, and GAPDH as internal control for TLR4 [ 21 ].…”

“…Cells were cultured at 37 °C in 95% humidity and 5% CO2. 6 different treatments which cell underwent are as follows: sevoflurane (Sev), negative control oligonucleotide (NC), miR-424 mimic (mimic), pLenti-GIII-CMV vector (pcDNA), and pLenti-GIII-TLR4 plasmid (TLR4) [ 21 ].…”