Open AccessCytokine gene expression in monocytes of patients undergoing cardiopulmonary bypass surgery evaluated by real‐time PCR
Author(s) -
Zimmermann Anja K.,
Simon P.,
Seeburger J.,
Hoffmann J.,
Ziemer G.,
Aebert H.,
Wendel H. P.
Publication year - 2003
Publication title -
journal of cellular and molecular medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.44
H-Index - 130
eISSN - 1582-4934
pISSN - 1582-1838
DOI - 10.1111/j.1582-4934.2003.tb00213.x
Subject(s) - cardiopulmonary bypass , proinflammatory cytokine , cardiac surgery , real time polymerase chain reaction , medicine , reverse transcriptase , gene expression , bypass grafting , messenger rna , coronary artery bypass surgery , cytokine , artery , monocyte , anesthesia , gene , immunology , andrology , rna , inflammation , surgery , biology , biochemistry
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‐β, 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.