Cytokine gene expression in monocytes of patients undergoing cardiopulmonary bypass surgery evaluated by real-time PCR.

Cardiopulmonary bypass (CPB) surgery induces systemic release of proinflammatory cytokines causing unspecific inflammatory reactions. This study deals with the development of a sensitive technique for detecting changes at the mRNA level in monocytes of patients undergoing CPB surgery, by using real-time PCR. Blood samples from patients undergoing elective coronary artery bypass grafting were obtained at six different time points. RNA was extracted from isolated monocytes and cDNA was synthesized by reverse transcriptase. CPB surgery induced gene expression of IL-1beta, IL-6, IL-8, and TNF-alpha, followed by a decrease below the preoperative expression values 6 h post CPB. High significant increases in gene expression for IL-8 at the end of surgery (p = 0.001) were detected. Real-time PCR is a powerful tool for getting simultaneously numerous sensitive, accurate, and reliable results from small amounts of biological material. This method avoids time-consuming and hazardous post-PCR manipulations and decreases the potential risk of PCR contamination.