Journal: Anesthesia & Analgesia

Article Title: Remifentanil Induces Cardio Protection Against Ischemia/Reperfusion Injury by Inhibiting Endoplasmic Reticulum Stress Through the Maintenance of Zinc Homeostasis

doi: 10.1213/ane.0000000000003414

Figure Lengend Snippet: Figure 2. Preconditioning with Rem rescues Zn2+ deficiency-induced ER stress in isolated ischemic reperfused rat hearts. Compared with the I/R group, Rem preconditioning reversed Zn2+ outflux from the heart (A) via inhibiting the expression of MTF1 (B and C) and its target gene ZnT1 (B and D). Rem attenuated ER stress by decreasing the expression of CHOP (E and F) and BIP (E and G), which were abolished by TPEN. *P < .05 versus sham; #P < .05 versus I/R; †P < .05 versus Rem + I/R. BIP indicates binding immunoglobulin protein; CHOP, CCAAT-enhancer-binding protein homolo- gous protein; ER, endoplasmic reticulum; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; I/R, ischemia/reperfusion; MTF1, metal-responsive transcription factor 1; Rem, remifentanil; TPEN, N,N,N′,N′-tetrakis-(2-pyridylmethyl) ethylenediamine; ZnT1, zinc transporter protein 1; Zn2+, zinc.

Article Snippet: The membranes were blocked with 5% nonfat milk solution and incubated with primary antibodies, including antiMTF1 (1:1000; Biobyte, Claremont, CA), anti-ZnT1 (1:1000; Invitrogen, Carlsbad, CA), anti-CCAAT-enhancer-binding protein homologous protein (CHOP) (1:1000; Cell Signaling Technology [CST], Beverly, MA), anti-binding immunoglobulin protein (BIP) (1:1000; CST), anti-B-cell lymphoma 2 (Bcl2) (1:1000; CST), anti-Bcl2-associated X protein (Bax) (1:1000; Abcam, Cambridge, MA), anti-Caspase3 (1:1000; CST), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:1500; CST) antibodies.

Techniques: Isolation, Expressing, Binding Assay

Journal: Anesthesia & Analgesia

Article Title: Remifentanil Induces Cardio Protection Against Ischemia/Reperfusion Injury by Inhibiting Endoplasmic Reticulum Stress Through the Maintenance of Zinc Homeostasis

doi: 10.1213/ane.0000000000003414

Figure Lengend Snippet: Figure 5. Effect of Rem on ZnT1 expression and ER stress in H9c2 cells overexpressed with MTF1. A, Confocal images of cells loaded with ER Tracker Red and Newport Green DCF. B, Western blotting images of ZnT1, CHOP, and BIP expressions. C–E, Summarized data for ZnT1, CHOP, and BIP expressions. *P < .05 versus control with vector; #P < .05 versus H/R with vector; †P < .05 versus H/R with vector; §P < .05 versus Rem + H/R with vector. Scale bar: 10 μm. BIP indicates binding immunoglobulin protein; CHOP, C/EBP homologous protein; DCF, dichlorodi- hydrofluoresceine; ER, endoplasmic reticulum; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; H/R, hypoxia/reoxygenation; H9c2, the embryonic rat heart–derived myogenic cell line H9c2; MTF1, metal-responsive transcription factor 1; Rem, remifentanil; Tag2B, pCMV-Tag2B; ZnT1, zinc transporter protein 1.

Article Snippet: The membranes were blocked with 5% nonfat milk solution and incubated with primary antibodies, including antiMTF1 (1:1000; Biobyte, Claremont, CA), anti-ZnT1 (1:1000; Invitrogen, Carlsbad, CA), anti-CCAAT-enhancer-binding protein homologous protein (CHOP) (1:1000; Cell Signaling Technology [CST], Beverly, MA), anti-binding immunoglobulin protein (BIP) (1:1000; CST), anti-B-cell lymphoma 2 (Bcl2) (1:1000; CST), anti-Bcl2-associated X protein (Bax) (1:1000; Abcam, Cambridge, MA), anti-Caspase3 (1:1000; CST), and anti-glyceraldehyde 3-phosphate dehydrogenase (GAPDH) (1:1500; CST) antibodies.

Techniques: Expressing, Western Blot, Control, Plasmid Preparation, Binding Assay, Derivative Assay