Introduction: Patients with chronic hepatitis C (CHC) have an increased risk of atherosclerotic cardiovascular disease which may be due to inflammation and endothelial dysfunction caused by the chronic infection. In this prospective pilot study, we assessed, for the first time among patients with CHC the myocardial perfusion reserve (MPR) by Rubidium-82 (⁸²Rb) positron emission tomography (PET)/computed tomography (CT) before and after direct-acting antiviral (DAA) treatment and compared them with biomarkers of systemic inflammation and endothelial dysfunction. Methods: We included 10 patients with CHC who received 8 or 12 weeks of DAA treatment. To obtain the MPR, a cardiac ⁸²Rb PET/CT scan at rest and adenosine-induced stress was performed at baseline and between 12 and 24 weeks post DAA treatment. Additionally, markers of endothelial dysfunction and inflammation were measured at baseline and 12 weeks after DAA treatment. Results: All 10 patients achieved cure and the median age was 50 (range: 40–62 years). The median MPR before treatment was 3.1 (range: 2.3–4.8) compared to 2.9 (range: 2.2–4.1) after DAA treatment p = 0.63. Also, cure after DAA treatment was not associated with an overall significant decrease in markers of endothelial dysfunction and inflammation. Discussion: Cure after DAA treatment in patients with CHC did not improve coronary microvascular function nor did it lead to a decrease in soluble markers of cardiovascular risk in the given time frame where the patients were followed. It should be noted, that MPR before DAA treatment was in the normal range. Considering the small sample size and short follow-up time, further studies are warranted to determine if viral clearance has an effect on coronary microvascular function and endothelial dysfunction.