A carrier liquid, the perfusate, is perfused into the probe.
In this perfusion system, the perfusate pumped into the abdominal aorta is circulated through one uterine artery– placenta fetus uterine vein unit and drips from the drain tube inserted in the caudal vein.
To examine the role of GLP-1 and PKA in the setting of the anti-diabetic gliptin drugs, Exendin 9 39 or DMSO (3 nM, a GLP-1 receptor antagonist, Sigma) were added into the perfusate in selected experiments and H-89 or DMSO (5 μM, a PKA antagonist, Sigma) was administered by intraperitoneal (I.P) injection prior to excision of the heart.
Tissue temperatures are monitored and radiolabeled serum albumen or erythrocytes are injected into the perfusate to detect leakage into the systemic circulation by a precordial scintillation probe.
While cumulative biliary excretion of unconjugated valerenic acid and the glucuronides M1 M7 dropped dramatically to 1 9%, their efflux into perfusate increased 1.5- to 10-fold.
To determine the RR, 5 μL of each dialysate and perfusate was pipetted into a counting vial containing 3 mL scintillation fluid (High-flash Point, Universal LSC-Cocktail, ULTIMA GOLD™, PerkinElmer , Inc.
Probe perfusate was collected into sterile CMA collection vials, transferred to reduced volume 300 μL polypropylene autosampler vials (AH0-7777, Phenomenex, Torrance, CA, USA) and stored at -20°C.
PC + NAC + NaHS group: The procedure was similar to that for group 5, except that 10 mM NAC were injected into perfusate simultaneously via a side arm for 10 min during early reperfusion (only aging rat hearts).
PC + LY294002 + NaHS group: The procedure was similar to that for group 5, except that 10 μM LY294002 were injected into perfusate simultaneously via a side arm for 10 min during early reperfusion (only aging rat hearts).
