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L. Tong, M. Cai, Y. Huang, H. Zhang, B. Su, Z. Li, H. Dong 

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Presentation on theme: "L. Tong, M. Cai, Y. Huang, H. Zhang, B. Su, Z. Li, H. Dong "— Presentation transcript:

1 Activation of K2P channel–TREK1 mediates the neuroprotection induced by sevoflurane preconditioning 
L. Tong, M. Cai, Y. Huang, H. Zhang, B. Su, Z. Li, H. Dong  British Journal of Anaesthesia  Volume 113, Issue 1, Pages (July 2014) DOI: /bja/aet338 Copyright © 2014 The Author(s) Terms and Conditions

2 Fig 1 Characterization of stimulated SH-SY5Y cells. (a–c) Western blot analysis for expression of the neuronal markers NeuN and βIII-tubulin in cultured SH-SY5Y cells stimulated by RA for 1, 4, 7, or 10 days. (a) Representative immunoblots. Histograms showing densitometric analyses of NeuN (b) and βIII-tubulin expression (c). Data presented as mean (sem). *P<0.05 compared with 1 day. (d) Photomicrographs of immunofluorescent staining showing expression of the two neuronal markers and TREK-1 in differentiated SH-SY5Y cells after RA stimulation for 7 days. Bar=20 μm. British Journal of Anaesthesia  , DOI: ( /bja/aet338) Copyright © 2014 The Author(s) Terms and Conditions

3 Fig 2 TREK-1 mediates sevoflurane preconditioning-induced neuroprotection in vitro. (a) Western blot analysis showing the efficiency of TREK-1 siRNA knockdown of the expression of TREK-1 in differentiated SH-SY5Y cells. Normal, untreated control cells; siRNA, cells treated with TREK-1 siRNA; siRNA-c, cells treated with AllStars Negative Control siRNA. Note that protein expression of TREK-1 was inhibited by TREK-1 siRNA compared with either the normal or siRNA-c group. Data presented as mean (sem). *P<0.05 compared with the normal control group. (b) Histograms showing cell viabilities in the normal (control), OGD, sevoflurane preconditioning plus OGD (Pre+OGD), TREK-1 siRNA plus sevoflurane preconditioning plus OGD (siRNA+Pre+OGD), or control siRNA plus sevoflurane preconditioning plus OGD (siRNA−c+Pre+OGD) groups. *P<0.05 vs OGD group. #P<0.05 vs Pre+OGD group. (c) Western blot analysis for caspase-3 expression. Data presented as mean (sem). *P<0.05 vs the OGD group. #P<0.05 compared with the Pre+OGD group. (d) Representative immunofluorescence images showing activation of caspase-3 in SH-SY5Y cells at 24 h after OGD. Bar=20 μm. British Journal of Anaesthesia  , DOI: ( /bja/aet338) Copyright © 2014 The Author(s) Terms and Conditions

4 Fig 3 Sevoflurane preconditioning induced neuroprotection in focal cerebral ischaemia. (a, d) Comparisons of neurobehavioral manifestations among the sham, MCAO, and pre+MCAO (sevoflurane preconditioning plus MCAO) groups at 24, 48, 72 h (a) and 7 days (d) after reperfusion. Each symbol represents the score for a single rat. A horizontal bar indicates the median value for each group (n=8–9 for each). *P<0.05 compared with the MCAO group. Representative photograph showing infarct volume in different groups 3 (b) and 7 days (e) after MCAO. Brain slices stained with TTC were from three representative rats subjected to sham, MCAO or sevoflurane preconditioning plus MCAO (Pre+MCAO). Histogram comparing infarct volume between the three groups at 3 (c) and 7 days (f) after reperfusion. Data presented as mean (sem) with individual values. *P<0.05 compared with the MCAO group (n=8–9). British Journal of Anaesthesia  , DOI: ( /bja/aet338) Copyright © 2014 The Author(s) Terms and Conditions

5 Fig 4 Sevoflurane preconditioning induced increase in TREK-1 protein and mRNA expression. (a) Western blot analysis of protein expression of TREK-1 in the sham, MCAO, and sevoflurane preconditioning plus MCAO (Pre+MCAO) groups at 4, 24, and 48 h after reperfusion. The upper panel shows TREK-1 and the corresponding β-actin bands. The lower panel histogram shows the results of the densitometric analysis. (b) mRNA expression of TREK-1 in the sham, MCAO, and sevoflurane preconditioning plus MCAO (Pre+MCAO) groups at 4, 24, and 48 h after reperfusion. *P<0.05 compared with the sham group. #P<0.05 compared with the MCAO group at the corresponding time point (n=8 for each). British Journal of Anaesthesia  , DOI: ( /bja/aet338) Copyright © 2014 The Author(s) Terms and Conditions

6 Fig 5 Knockdown of TREK-1 expression attenuates the neuroprotective effect induced by sevoflurane preconditioning in the rat MCAO model. (a) Western blot and real-time PCR analysis showing the effect of siRNA on TREK-1 protein expression and mRNA in sham (untreated control rats), siRNA (rats injected with TREK-1 siRNA), and negative (rats treated with AllStars Negative Control siRNA). The upper panel shows the protein expression of TREK-1. The lower panel shows TREK-1 mRNA expression relative to GAPDH. *P<0.05 compared with the sham group. (b) Effect of TREK-1 siRNA on infarct volume. Brain slices stained with TTC from five representative rats chosen for each group. (c) Comparisons of infarct volumes. *P<0.05 compared with the MCAO group. #P<0.05 compared with the Pre+MCAO group. (d) Effect of TREK-1 siRNA on neurobehavioural indices. Each symbol represents the score for a single rat. A horizontal bar indicates the mean for each group. *P<0.05 compared with the MCAO group. #P<0.05 compared with the Pre+MCAO group (n=8 for each). British Journal of Anaesthesia  , DOI: ( /bja/aet338) Copyright © 2014 The Author(s) Terms and Conditions

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