HIV protease inhibitor ritonavir increases endothelial monolayer permeability.
HIV protease inhibitors (PIs) are often associated with metabolic and cardiovascular complications although they are effective anti-HIV drugs. In this study, we determined whether HIV PI ritonavir could increase endothelial permeability, one of the important mechanisms of vascular lesion formation. Human dermal microvascular endothelial cells (HMECs) treated with ritonavir showed a significant increase of endothelial permeability in a dose- and time-dependent manner assayed with a transwell system. Ritonavir significantly reduced the mRNA levels of tight junction proteins zonula occluden-1, occludin, and claudin-1 by 40-60% as compared to controls (P<0.05) by real-time PCR analysis. Protein levels of these tight junction molecules were also substantially reduced in the ritonavir-treated cells. In addition, HMECs treated with ritonavir (7.5, 15, and 30microM) showed a substantial increase of superoxide anion production by 10%, 32%, and 65%, respectively, as compared to controls. Antioxidants (EGCG and SeMet) effectively reduced ritonavir-induced endothelial permeability. Furthermore, ritonavir activated ERK1/2 (phosphorylation), but not P38 and JNK. Specific ERK1/2 inhibitor, PD89059, significantly abolished ritonavir-induced endothelial permeability by 92%. Thus, HIV PI ritonavir increases endothelial permeability, decreases levels of tight junction proteins, and increases superoxide anion production. ERK1/2 activation is involved in the signal transduction pathway of ritonavir-induced endothelial permeability.
Chen
Changyi
C
Molecular Surgeon Research Center, Division of Vascular Surgery and Endovascular Therapy, Michael E. DeBakey Department of Surgery, Baylor College of Medicine, Houston, TX 77030, USA. jchen@bcm.tmc.edu
Lu
Xiang-Huai
XH
Yan
Shaoyu
S
Chai
Hong
H
Yao
Qizhi
Q
D015703:Antigens, CD; D001483:Base Sequence; D015820:Cadherins / Q000494:pharmacology; D002463:Cell Membrane Permeability / Q000187:drug effects*; D017931:DNA Primers; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects* / Q000201:enzymology; D017320:HIV Protease Inhibitors / Q000494:pharmacology*; D006801:Humans; D020928:Mitogen-Activated Protein Kinases / Q000378:metabolism; D018384:Oxidative Stress; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000235:genetics; D019438:Ritonavir / Q000494:pharmacology*; D013481:Superoxides / Q000378:metabolism; D019108:Tight Junctions; D000099152:Cadherin 5
D015703:Antigens, CD; D015820:Cadherins; D017931:DNA Primers; D017320:HIV Protease Inhibitors; D020928:Mitogen-Activated Protein Kinases; D012333:RNA, Messenger; D019438:Ritonavir; D013481:Superoxides; D000099152:Cadherin 5