Ischemic preconditioning (PC) is a polygenic defensive cellular adaptive phenomenon of the heart to ischemic stress, whereby the heart changes its phenotype to become more resistant to subsequent ischemia. Early and late of PC represent two chronologically and pathophysiologically distinct phases of this phenomenon, which can be recruited pharmacologically. We represent a post hoc analysis examining the late PC-mimetic effects of nitroglycerin (NTG) on peri-procedural myocardial necrosis during percutaneous coronary intervention (PCI). A group of 66 patients presenting with angina were randomized, 24 h prior to a scheduled PCI for single obstructive CAD, to a 4 h pretreatment with intravenous NTG or saline. Measurements of electrocardiographic ST-segment shifts, echocardiographic regional wall motion and angina scores demonstrated that NTG pre-treatment preconditioned the heart by rendering it resistant to ischemia during balloon inflations. NTG-pretreated patients exhibited trends towards lower average peak CK (131.1 vs. 188.6 U/L, P = 0.38) and CK-MB levels (7.1 vs. 12.6 ng/ml, P = 0.40). NTG, however, had no significant impact on the incidence of post-procedural MI or any cardiac enzyme elevation. The exploitation of ischemic and pharmacological PC may prove a useful strategy to confer cardioprotection during high-risk PCI and is worth exploring.