Remifentanil Preconditioning Reduces Postischemic Myocardial Infarction and Improves Left Ventricular Performance via Activation of the Janus Activated Kinase-2/Signal Transducers and Activators of Transcription-3 Signal Pathway and Subsequent Inhibition of Glycogen Synthase Kinase-3β in Rats
Authors: Xiaowen Mao
Shigang Qiao
Yan Wang
Shaoqing Lei
Gordon T Wong
Zhengyuan Xia
Tingting Wang
Chi-Wai Cheung
Michael G Irwin
Yanan Liu
Journal: Critical Care Medicine (2016)
Discipline: CLINICAL MEDICINE
Keywords: Remifentanil preconditioning; JAK2/STAT3 signaling; PI3K/Akt signaling; glycogen synthase kinase-3β; myocardial ischemia reperfusion injury
Affiliations: 1Department of Anesthesiology, the University of Hong Kong, Hong Kong SAR, China.2Department of Anesthesiology and Critical Care, the Second Affiliated Hospital of Soochow University, Suzhou, China.3Department of Anesthesiology, the Second Affiliated Hospital & Yuying Children's Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.4State Key Laboratory of Pharmaceutical Biotechnology, University of Hong Kong, Hong Kong, China.

Abstract:  Objectives: Remifentanil preconditioning attenuates myocardial ischemia reperfusion injury, but the underlying mechanism is incompletely understood. The Janus activated kinase-2 (JAK2)/signal transducers and activators of transcription-3 (STAT3) and phosphatidylinositol 3-kinase (PI3K)/Akt signaling pathways are critical in both ischemic and pharmacologic preconditioning cardioprotection, which involve the inactivation of glycogen synthase kinase-3β. We hypothesized that remifentanil preconditioning confers cardioprotection via the JAK2/STAT3 and/or PI3K/Akt activation-mediated glycogen synthase kinase-3β inhibition. Design: Pharmacologic intervention. Setting: Research laboratory. Subjects: Male Sprague-Dawley rats. Interventions: In vivo and in vitro treatments. Measurements and main results: Male Sprague-Dawley rats (n = 6 per group) were sham operated or subjected to myocardial ischemia reperfusion injury. The JAK2 inhibitor AG490 (3 mg/kg), the PI3K inhibitor wortmannin (15 μg/kg), or the glycogen synthase kinase-3β inhibitor SB216763 (600 μg/kg) were given before inducing in vivo myocardial ischemia reperfusion injury achieved by occluding coronary artery for 30 minutes followed by 120 minutes of reperfusion in the absence or presence of remifentanil preconditioning (6 μg/kg/min). Also, isolated rat hearts were Langendorff perfused and subjected to 30 minutes of global ischemia and 120 minutes of reperfusion without or with remifentanil preconditioning (100 ng/mL) in the presence or absence of AG490 and/or SB216763. Isolated rat cardiomyocytes and H9C2 cells were subjected to hypoxia/reoxygenation alone or in combination with AG490 (100 μM), wortmannin (100 nM), or SB216763 (3 μM) without or with remifentanil preconditioning (2.5 μM). Remifentanil preconditioning reduced postischemic myocardial infarction and hemodynamic dysfunction induced by myocardial ischemia reperfusion injury concomitant with increased phosphorylation of STAT3 at tyr-705 (p-STAT3) and glycogen synthase kinase-3β but not Akt. AG490 but not wortmannin cancelled remifentanil preconditioning cardioprotection, and SB216763 restored it despite the presence of AG490. In Langendorff-perfused hearts, AG490-mediated cancellation of remifentanil preconditioning cardioprotection in attenuating postischemic myocardial infarction and creatinine kinase-MB release was reverted by concomitant administration of SB216763. Remifentanil preconditioning also attenuated posthypoxic cardiomyocyte injury and increased p-STAT3 and glycogen synthase kinase-3β in isolated primary cardiomyocytes and H9C2 cells. STAT3 gene knockdown with specific synthetic RNA cancelled remifentanil preconditioning cardioprotection, whereas glycogen synthase kinase-3β gene knockdown, which per se did not affect STAT3 under hypoxia/reoxygenation condition, preserved remifentanil preconditioning cardioprotection regardless of STAT3 abrogation. Conclusions: Remifentanil preconditioning confers cardioprotection primarily via activation of JAK2/STAT3 signaling that can function independent of PI3K/Akt activation. Glycogen synthase kinase-3β is a critical downstream effector of remifentanil preconditioning cardioprotection. 

Retracted: No

PubPeer comments:
 Figure 5B. * Blue boxes: the RPC+I R and the AG+RPC+I R panels appear to show an overlap. The latter panel is shown darker and has a higher measured value in the bar graph below. ![file] , 
 The two images pointed out belong to the AG+RPC+I R group. The images were taken (at 400x amplification) from the same slide and same area with minor adjustment of the background brightness and field for us and the journal to have the option to decide which one was more representative or of better quality (visibility) for that particular group. However, it appears that an error has occurred at the last stage in that we have mistakenly used the same similar image for two different groups. **How did this error occur?** After reviewing the file folder of representative images, we note that we named the images for the RPC+I R group (as shown in the paper and figure) as Rem (note: this is an abbreviation of Remifentanil; with Rem1, Rem2 u2026to indicate different images within this group)uff0cand named the images for the AG+RPC+I R group (as shown in the paper and figure) as Rem+ (with Rem1+, Rem2+ u2026to indicate different images within this group) (note: one of the reasons that we used this nomenclature is that our stained slides were processed by an independent researcher (whose name was not in the author list) who was more experienced in apoptotic cell counting and image photo-taking at that time, and to reduce bias we did want to have an indicator of what Rem + meant (i.e. higher dose of Rem or Rem plus any inhibitor) u2013 we have mentioned this in the manuscript. Unfortunately, due to the similarity in image file names as well as our oversightuff0cwhen we chose a representative image for the RPC+I R group using an image with the file name Rem, we selected a Rem+ imageuff0cand as such, two images from the AG+RPC+I R group ended up being used to represent both the RPC+I R and AG+RPC+I R groups with apparent overlap within these two images, as they were taken from the same field. Details as followsuff1auff08NOTE: image data could not be displayed in this dialog box) RPC+I R groupuff08Rem1, image as aboveuff09 should use the above image, but by error used the file named Rem1+ u3002Consequently Both Rem1+ and Rem2+ appeared as presented in the published paper, to represent two different groups (i.e. the RPC+I R group and AG+RPC+I R group), despite both of them belonging to the AG+RPC+I R group and only one should have been used. The corrected Fig. is presented below. uff1a The mis-match in the representative image has no impact on the data information as presented in the accompanying bar graph. There was a statistically significant difference between the two groups concerned (as shown in the paper) and, thus, the mistaken use of the same image (in terms of degree of cell damage) does not represent the different degrees of cell damages as shown in the bar graph. In other words, the error reduced the quality of our figure. We will write to the journal to clarify this and send them the corrected figure. We will also acknowledge your assistance in pointing out this error. , 
 Thank you, Dr. Irwin, for your prompt reply and detailed analysis of what has happened. Much appreciated and I look forward to see the correction. , 
 Yes, if you don t mind could you DM your e-mail address to mgirwin@hku.hk. We will cc you the erratum. Best regards ,