G. Xu, H. Lu, Y. Dong, D. Shapoval, S. G. Soriano, X. Liu, Y. Zhang, Z

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Coenzyme Q10 reduces sevoflurane-induced cognitive deficiency in young mice G. Xu, H. Lu, Y. Dong, D. Shapoval, S.G. Soriano, X. Liu, Y. Zhang, Z. Xie British Journal of Anaesthesia Volume 119, Issue 3, Pages 481-491 (September 2017) DOI: 10.1093/bja/aex071 Copyright © 2017 The Author(s) Terms and Conditions

Fig 1 Diagram of experimental design. Young mice (n=134) received anaesthesia with 3% sevoflurane for 2 h or control on 3 days at postnatal day (P) 6, P7, and P8. We harvested brain tissue (hippocampus and cortex) at the end of anaesthesia on P8 for western blotting (n=24), measurement of adenosine triphosphate (ATP; n=36), and measurement of reactive oxygen species (ROS) and mitochondrial membrane potential (MMP; n=24). Different groups of mice (n=50) were used for assessment of learning and memory function using the Morris water maze from P31 to P37 after the birth. We harvested hippocampus at the end of the Morris water maze assessments (P37) for western blotting (n=50). British Journal of Anaesthesia 2017 119, 481-491DOI: (10.1093/bja/aex071) Copyright © 2017 The Author(s) Terms and Conditions

Fig 2 Coenzyme Q10 (CoQ10) attenuated sevoflurane-induced cognitive deficiency. (A) There was a non-significant interaction between group (control vs sevoflurane) and time [postnatal day (P) 31–P37] in the escape latency of the Morris water maze (MWM) test with pretreatment of corn oil (CoQ10 vehicle). (B) There was a significant difference in platform-crossing times between control mice and and mice exposed to sevoflurane anaesthesia. (C) There was not a significant interaction between group (control vs sevoflurane) and time (P31–P37) in MWM escape latency for pretreatment with CoQ10. (D) There was not a significant difference in platform-crossing times between control mice and the mice exposed to sevoflurane anaesthesia and pretreatment with CoQ10. n=10–16 in each group. British Journal of Anaesthesia 2017 119, 481-491DOI: (10.1093/bja/aex071) Copyright © 2017 The Author(s) Terms and Conditions

Fig 3 Coenzyme Q10 (CoQ10) attenuated the sevoflurane-induced reduction in postsynaptic density protein-95 (PSD-95) in hippocampus. (A) Anaesthesia with 3% sevoflurane for 2 h daily for 3 days decreased PSD-95 concentrations in the hippocampus compared with control conditions at postnatal day (P) 8. Coenzyme Q10 attenuated the sevoflurane-induced reduction in PSD-95 in hippocampus compared with corn oil at P8. (B) Quantification of western blot data showed that sevoflurane anaesthesia reduced PSD-95 concentrations in hippocampus compared with control conditions. Coenzyme Q10 attenuated the sevoflurane-induced reduction in PSD-95 in hippocampus compared with corn oil. (C) Western blotting showed that anaesthesia with 3% sevoflurane for 2 h daily for 3 days at P6, P7, and P8 decreased PSD-95 concentrations in hippocampus compared with control at P37. Coenzyme Q10 attenuated sevoflurane-induced reduction in PSD-95 in hippocampus compared with corn oil at P37. (D) Quantification of western blot data showed that the sevoflurane anaesthesia reduced PSD-95 in hippocampus compared with control conditions at P37. Coenzyme Q10 attenuated the sevoflurane-induced reduction in PSD-95 in hippocampus compared with corn oil at P37. n=6 in each group. British Journal of Anaesthesia 2017 119, 481-491DOI: (10.1093/bja/aex071) Copyright © 2017 The Author(s) Terms and Conditions

Fig 4 Coenzyme Q10 (CoQ10) attenuated the sevoflurane-induced reduction in synaptophysin in the hippocampus. (A) Western blotting showed that anaesthesia with 3% sevoflurane 2 h daily for 3 days decreased synaptophysin concentrations in hippocampus compared with control conditions at postnatal day (P) 8. Coenzyme Q10 attenuated the sevoflurane-induced decrease in synaptophysin in hippocampus compared with corn oil at P8. (B) Quantification of western blot data showed that sevoflurane anaesthesia reduced synaptophysin concentrations in hippocampus compared with control conditions at P8. Coenzyme Q10 attenuated the sevoflurane-induced reduction in synaptophysin in hippocampus compared with corn oil at P8. (C) Western blotting showed that anaesthesia with 3% sevoflurane for 2 h daily for 3 days at P6, P7, and P8 decreased synaptophysin in hippocampus compared with control conditions at P37. Coenzyme Q10 attenuated the sevoflurane-induced decrease in synaptophysin in hippocampus compared with corn oil at P37. (D) Quantification of western blot data showed that sevoflurane anaesthesia reduced synaptophysin in hippocampus compared with control conditions at P37. Coenzyme Q10 attenuated the sevoflurane-induced reduction in synaptophysin in hippocampus compared with corn oil at P37. n=6 in each group. British Journal of Anaesthesia 2017 119, 481-491DOI: (10.1093/bja/aex071) Copyright © 2017 The Author(s) Terms and Conditions

Fig 5 Coenzyme Q10 (CoQ10) attenuated the sevoflurane-induced reduction in adenosine triphosphate (ATP) in cerebral cortex. Anaesthesia with 3% sevoflurane for 2 h daily for 3 days [postnatal day (P) 6, P7, and P8] decreased amounts of ATP in cortex compared with control conditions at P8. Coenzyme Q10 attenuated the sevoflurane-induced reduction in ATP in cortex compared with corn oil at P8. n=9 in each group. British Journal of Anaesthesia 2017 119, 481-491DOI: (10.1093/bja/aex071) Copyright © 2017 The Author(s) Terms and Conditions

Fig 6 Coenzyme Q10 (CoQ10) did not attenuate the sevoflurane-induced increase in reactive oxygen species (ROS) in hippocampus. Anaesthesia with 3% sevoflurane for 2 h daily for 3 days [postnatal day (P) 6, P7, and P8] increased amounts of ROS in hippocampus compared with the control conditions at P8. Coenzyme Q10 did not attenuate the sevoflurane-induced increase in ROS in hippocampus compared with corn oil. n=6 in each group. British Journal of Anaesthesia 2017 119, 481-491DOI: (10.1093/bja/aex071) Copyright © 2017 The Author(s) Terms and Conditions

Fig 7 Coenzyme Q10 (CoQ10) attenuated the sevoflurane-induced reduction in mitochondrial membrane potential (MMP) in cerebral cortex. Anaesthesia with 3% sevoflurane for 2 h daily for 3 days [postnatal day (P) 6, P7, and P8] decreased MMP in cortex compared with control conditions at P8. Coenzyme Q10 attenuated the sevoflurane-induced reduction in MMP in cortex compared with corn oil. n=6 in each group. British Journal of Anaesthesia 2017 119, 481-491DOI: (10.1093/bja/aex071) Copyright © 2017 The Author(s) Terms and Conditions