Journal: Anesthesiology

Article Title: Free Fatty Acid Receptor GPR40 Mediates Lipid Emulsion-induced Cardioprotection

doi: 10.1097/ALN.0000000000002195

Figure Lengend Snippet: Heart Rate (A) and EF (B) before and 30 min after GW1100 (n=7). C. M-Mode Transthoracic echocardiographic images from CTRL (lipid emulsion only, upper panel) and lipid emulsion+GPR40 antagonist (lower panel) at baseline, and 1, 5, 10 min after lipid emulsion. Heart rate (D) and EF (E) at baseline, and 1, 5, 10 min after lipid emulsion (n=4-7/group, ****p<0.0001; **p<0.01; NS: not significant). LE, lipid emulsion; BUP, bupivacaine; EF, ejection fraction. Values are expressed as mean ± SD.

Article Snippet: Protein concentration was measured and 100 μg of total protein was loaded on a 4-20% gradient Tris-HCl/SDS polyacrylamide gel, electrotransferred to nitrocellulose paper, blocked with 5% non-fat dry milk in 20 mM TBS with 0.1% Tween, and incubated with primary GPR40 antibody (SC-32905, Santa Cruz Biotechnology, Dallas, TX, USA, 1:1,000 dilution) and Vinculin antibody (Sigma cat# V3191, 1:20,000) overnight at 4°C.

Techniques: Emulsion

Journal: Anesthesiology

Article Title: Free Fatty Acid Receptor GPR40 Mediates Lipid Emulsion-induced Cardioprotection

doi: 10.1097/ALN.0000000000002195

Figure Lengend Snippet: A) Western blot of the heart lysates from mice (left) and rats (right) showing a single band at the expected molecular weight for GPR40 (arrow) and vinculin (arrow). B) Immunofluorescence stained representative isolated mouse heart cardiomyocyte showing GPR40 (red) and DAPI (blue) labeling.

Article Snippet: Protein concentration was measured and 100 μg of total protein was loaded on a 4-20% gradient Tris-HCl/SDS polyacrylamide gel, electrotransferred to nitrocellulose paper, blocked with 5% non-fat dry milk in 20 mM TBS with 0.1% Tween, and incubated with primary GPR40 antibody (SC-32905, Santa Cruz Biotechnology, Dallas, TX, USA, 1:1,000 dilution) and Vinculin antibody (Sigma cat# V3191, 1:20,000) overnight at 4°C.

Techniques: Western Blot, Molecular Weight, Immunofluorescence, Staining, Isolation, Labeling

Journal: Anesthesiology

Article Title: Free Fatty Acid Receptor GPR40 Mediates Lipid Emulsion-induced Cardioprotection

doi: 10.1097/ALN.0000000000002195

Figure Lengend Snippet: A. Left ventricular developed pressure (LVDP, mmHg); B. Rate pressure product (RPP, mmHg*beats/min); C. dP/dt max (mmHg/s), D. dP/dt min (mmHg/s), from Langendorff perfused mouse hearts in CTRL (n=6), lipid emulsion (LE, n=8) and lipid emulsion+GPR40 antagonist (n=7) with I/R injury as a function of time before ischemia, during ischemia, and reprfusion. ****p<0.0001; ***p<0.001; **p<0.01; *p<0.05; ####p<0.0001; ###p<0.001; ##p<0.01; #p<0.05; $p<0.05. Values are expressed as mean ± SD.

Article Snippet: Protein concentration was measured and 100 μg of total protein was loaded on a 4-20% gradient Tris-HCl/SDS polyacrylamide gel, electrotransferred to nitrocellulose paper, blocked with 5% non-fat dry milk in 20 mM TBS with 0.1% Tween, and incubated with primary GPR40 antibody (SC-32905, Santa Cruz Biotechnology, Dallas, TX, USA, 1:1,000 dilution) and Vinculin antibody (Sigma cat# V3191, 1:20,000) overnight at 4°C.

Techniques: Emulsion

Journal: Anesthesiology

Article Title: Free Fatty Acid Receptor GPR40 Mediates Lipid Emulsion-induced Cardioprotection

doi: 10.1097/ALN.0000000000002195

Figure Lengend Snippet: Glycogen synthase kinase-3 beta (GSK-3β) phosphorylation-induced inhibition of the mitochondrial permeability transition pore (mPTP) is likely to be the common downstream cardioprotective mechanism in both ischemia/reperfusion injury(1,3) and bupivacaine-induced cardiotoxicity(8,9) involving reperfusion injury salvage kinase (RISK) and/or survival activating factor enhancement (SAFE) pathways. Lipid emulsion can also result in increased adenosine triphosphate (ATP) production. GW stands for GPR40 antagonist GW1100.

Article Snippet: Protein concentration was measured and 100 μg of total protein was loaded on a 4-20% gradient Tris-HCl/SDS polyacrylamide gel, electrotransferred to nitrocellulose paper, blocked with 5% non-fat dry milk in 20 mM TBS with 0.1% Tween, and incubated with primary GPR40 antibody (SC-32905, Santa Cruz Biotechnology, Dallas, TX, USA, 1:1,000 dilution) and Vinculin antibody (Sigma cat# V3191, 1:20,000) overnight at 4°C.

Techniques: Phospho-proteomics, Inhibition, Permeability, Emulsion