ABSTRACT
Objective:To observe the effects of milrinone on LPS-induced inflammation and NF-κB activation of human coronary artery endothelial cells (HCAEC).Methods:HCAEC were cultured in vitro,and were divided into normal group,LPS group (1 μg/ml),milrinone 50 μmol/L group and milrinone 200 μmol/L group,the rear two groups were co-incubated with LPS for 24 h. qRT-PCR and ELISA was applied to detect IL-6,IL-8,TNF-α mRNA and protein,respectively.Western blot and immunofluorescence assay was used to observe the activation and translocation of NF-κB,respectively.Results: Compared with LPS group,the IL-6,IL-8, TNF-α mRNA and protein;the activation and translocation of NF-κB significantly inhibited after incubation with milrinone (P<0.05) in a dose dependent manner.Conclusion:Milrinone inhibited the release of inflammative cytokines induced by LPS of HCAEC in vitro, partly by inactivation of NF-κB.
ABSTRACT
During the last two decades, there has been an increasing interest in the impact of oral health on atherosclerosis and subsequent cardiovascular disease (CVD). To date, some periodontal pathogens including Porphyromonas gingivalis (P. gingivalis) have been reported to be relevant to CVD. Porphyromonas endodontalis (P. endodontalis), which shares approximately 87% sequence homology with P. gingivalis, is mostly found within infected root canals. However, recent studies reveal that this pathogen also resides in the dental plaque or periodontal pocket in patients with periodontitis. It has been shown that P. endodontalis invades human coronary artery endothelial cells (HCAEC) and coronary artery smooth muscle cells (CASMC). To evaluate whether P. endodontalis can participate in the progression of atherosclerosis and CVD, we examined the changes in transcriptional gene expression profiles of HCAEC responding to invasion by P. endodontalis in this study. The following results were obtained. 1. Porphyromonas endodontalis was invasive of HCAEC. 2. According to the microarray analysis, there were 625 genes upregulated more than two-folds, while there were 154 genes downregulated by half. 3. Upregulated genes were relevant to inflammatory cytokines, apoptosis, coagulation and immune response. Enhanced expression of MMP-1 was also noticeable. 4. The transcription profiles of the 10 selected genes examined by real-time PCR agreed well with those observed in the microarray analysis. Thus, these results show that P. endodontalis presents the potential to trigger and augment atherosclerosis leading to CVD.