Activation and Overexpression of PARP-1 in Circulating Mononuclear Cells Promote TNF-α and IL-6 Expression in Patients with Unstable Angina

Background: Proinflammatory cytokines are involved in the development of unstable angina (UA). Poly(ADP-ribose) polymerase-1 (PARP-1) contributes importantly to regulating the transcription of inflammatory cytokines. This study aims to investigate the relationship of PARP-1 in circulating mononuclear cells (MNCs) and plasma TNF-α and IL-6 in UA patients and to elucidate the mechanism that PARP-1 promotes TNF-α and IL-6 expression via NF-κB pathway. Methods: Twenty six Braunwald class IIIB UA patients, 25 stable angina patients and 25 healthy volunteers were enrolled in this study. Plasma TNF-α and IL-6 were determined with ELISA. Circulating MNCs were analyzed for PARP activity, PARP-1 expression and NF-κB DNA binding activity. MNCs from healthy subjects were cultured to investigate the direct effects of PARP-1 on NF-κB DNA binding activity and the expression of TNF-α and IL-6. Results: PARP activity and PARP-1 expression in circulating MNCs were increased and positively correlated with plasma TNF-α and IL-6, respectively, in UA patients. Spontaneous NF-κB activation in MNCs was demonstrated in UA patients. In cultured MNCs from healthy subjects, inhibition of PARP-1 prevented lipopolysaccharide-induced increase in DNA binding activity of NF-κB and the expression of TNF-α and IL-6. Supershift assay demonstrated that PARP-1 was a component of NF-κB/DNA complex. Addition of recombinant human PARP-1 protein to nuclear extracts of MNCs significantly increased the DNA binding activity of NF-κB. Conclusions: Activation and overexpression of PARP-1 are demonstrated in circulating MNCs of UA patients. Overexpressed PARP-1 promotes PARP-1/NF-κB/DNA complex formation, thereby enhancing the expression of TNF-α and IL-6 in circulating MNCs of UA patients.