Azithromycin reduces Chlamydia pneumoniae-induced attenuation of eNOS and cGMP production by endothelial cells
Authors: Bouwman, J. J. M.; Visseren, F. L. J.1; Bevers, L. M.1; van der Vlist, W. E.2; Bouter, K. P.3; Diepersloot, R. J. A.2
Source: European Journal of Clinical Investigation, Volume 35, Number 9, September 2005 , pp. 573-582(10)
Publisher: Blackwell Publishing
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Abstract:
Background Intracellular infections with cytomegalovirus (CMV) or Chlamydia pneumoniae (Cp) may play a role in the aetiology of atherosclerosis. Nitric oxide (NO) is a key regulator of endothelial function. Under pathological conditions uncoupling of endothelial nitric oxide synthase (eNOS) leads to vessel damage as a result of production of oxygen radicals instead of NO. We hypothesized that infection-induced atherosclerosis is initiated by changes in NO metabolism and may be reversed by azithromycin treatment. Methods Confluent human umbilical vein endothelial cells (HUVECs) were infected with Cp or CMV. After 48 h of infection, production of eNOS, cyclic guanosine monophosphate (cGMP) and reactive oxygen species (ROS) was measured. Detection of cGMP was used as a reporter assay for the bioavailability of NO. Subsequently, Cp- and CMV-infected HUVECs were coincubated with 0·016 mg L-1 and 1 mg L-1 azithromycin. Results Infection with Cp (MOI 1 and MOI 0·1) and CMV (MOI 1) caused a dose- and time-dependent reduction of eNOS production in the HUVECs: Cp MOI 1: 1141 ± 74 pg mL-1 (P < 0·01); Cp MOI 0·1: 3189 ± 30 pg mL-1 (P < 0·01); CMV: 3213 ± 11 pg mL-1 (P < 0·01) vs. 3868 ± 83 pg mL-1 for uninfected HUVECs. Chlamydia pneumoniae– but not CMV-infection also reduced cGMP-production (Cp: 0·195 ± 0·030 pmol mL-1 (P < 0·01); CMV: 0·371 ± 27 pmol mL-1 (P > 0·05) vs. 0·378 ± 0·019 pmol mL-1 for uninfected HUVECs). CMV-infection did not affect ROS production either, but Cp-infection reduced ROS-production by 21% (P > 0·05; Cp MOI 0·1) to 68% (P < 0·01; Cp MOI 1). Azithromycin treatment restored Cp-induced eNOS, cGMP and ROS production in a dose-dependent manner. Conclusions Infection with Cp in endothelial cells in vitro attenuates eNOS, cGMP and ROS production in HUVECs and azithromycin reverses Cp-induced effects on eNOS, cGMP and ROS-production. The results from our in vitro research support the role of antibiotic therapy for infection-induced atherosclerosis by indicating that azithromycin does actually improve endothelial function. Eur J Clin Invest 2005; 35 (9): 573 –582Keywords: Chlamydia pneumoniae; cyclic guanosine monophosphate; cytomegalovirus; endothelial dysfunction; endothelial nitric oxide synthase; reactive oxygen species
Document Type: Research article
DOI: 10.1111/j.1365-2362.2005.01541.x
Affiliations: 1: University Medical Center Utrecht, Utrecht, and 2: Diakonessen Hospital Utrecht, 3: Bosch Medicentre, ‘s Hertogenbosch, the Netherlands
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