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Ritonavir and Lopinavir Boosted with Ritonavir
Induce Endothelial dysfunction and Premature Senescence in Cultured Human Coronary Artery Endothelial Cells
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Reported by Jules Levin
CROI 2010 Feb 16-19 SF
C Lefevre1, M Auclair1, E Capel1, C Vigouroux1,2, Jacqueline Capeau*1,2, F Boccara1,2,3, and M Caron-Debarle1
1INSERM U938, UMRS938, Univ Paris 6 and Pierre and Marie Curie, France; 2Hosp Tenon, AP-HP, Paris, France; and 3Hosp Saint Antoine, AP-HP, Paris, France
AUTHOR CONCLUSION
Long-term exposure of human coronary artery endothelial cells to RTV and LPV/r progressively induced endothelial dysfunction at di_erent levels : altered production of NO and adhesion molecules, increased oxidative stress and inflammation .
Endothelial cell dysfunction was associated with increased expression of the senescence protein prelamin A and signs of early cellular senescence.
We suggest that PI-induced endothelial dysfunction might result in stress-induced premature cell senescence.
This could participate to early cardio-vascular diseases and aging occurring in some PI-treated patients.
ABSTRACT
Background: Clinical studies have shown that treatment with some protease inhibitors is associated with atherogenic lipid profiles and endothelial dysfunction and could result in early cardiovascular diseases. Studies in vitro demonstrated that short-term treatment with some protease inhibitors could directly alter endothelial function. However, the long-term effect of protease inhibitors on endothelial function and senescence has not been evaluated.
Methods: We studied the sequential impact on endothelial function and senescence of a chronic incubation of human coronary artery endothelial cells with ritonavir (RTV, 7.5 mM) or lopinavir (10 mM) plus ritonavir (2 mM) (RTV/r). Endothelial cell dysfunction was assessed at 10, 20, and 30 days, by nitric oxide (NO) production (DAF fluorescence), level of oxidative stress (CM-H2DCFDA oxidation, NBT reduction), inflammation (IL-6, IL-8, MCP-1, PAI-1 secretion) and production of adhesion proteins (VCAM, ICAM). Proliferation, replication and senescence were measured by the population doubling level, BrdU incorporation into DNA, senescence-associated b-galactosidase activity and altered cell morphology. The level of the senescence marker prelamin A was also determined.
Results: Long-term treatment with RTV and LPV/r resulted in progressive endothelial cell dysfunction with increased oxidative stress (3 to 12 fold), decreased NO production (2 to 3 fold) and protein expression of NO synthase 3 (2.5 fold). The secretion of IL-6, IL-8, and MCP-1 was markedly increased after chronic protease inhibitor treatment, as well as the secretion of PAI-1 (2 to 3.5 fold), ICAM and VCAM (2 to 7 fold). The 2 protease inhibitors decreased cell proliferation (by 2 to 3 fold) and replication (4 to 5 fold). They also progressively induced cellular senescence with increased SA-b-galactosidase activity (5 to 7 fold), altered cell morphology, and increased prelamin A accumulation (10 to 12 fold).
Conclusions: Long-term exposure of human coronary artery endothelial cells with RTV and LPV/r progressively induced endothelial dysfunction at different levels with decreased NO production, increased oxidative stress and secretion of inflammation and adhesion molecules. Cell dysfunction was associated with increased expression of the senescence protein prelamin A and signs of early cellular senescence. We suggest that RTV and LPV/r-induced endothelial dysfunction might result in stress-induced premature cell senescence. This could participate to early cardiovascular diseases and aging occurring in some protease inhibitor-treated patients.
BACKGROUND
Clinical studies have shown that treatment with some protease inhibitors is associated with atherogenic lipid profiles and endothelial dysfunction and could result in early cardiovascular diseases. In vitro studies demonstrated that short-term treatment with some PIs could directly alter endothelial function. However, the long-term effect of PIs on endothelial function and senescence has not been evaluated.
Methods
We studied the sequential impact on endothelial function and senescence of a chronic incubation of human coronary artery endothelial cells with ritonavir
(RTV, 7.5 microM) or lopinavir (10 microM) plus ritonavir (2 microM) (LPV/r). Endothelial cell dysfunction was assessed at 10, 20 and 30 days, by nitric oxide
(NO) production (DAF fluorescence), oxidative stress (CM-H2DCFDA oxidation, NBT reduction), inflammation (IL-6, IL-8, MCP-1 secretion) and production of
adhesion proteins (sVCAM, sICAM) and PAI-1. Proliferation, replication and senescence were measured by the population doubling level, BrdU incorporation
into DNA, senescence-associated beta-galactosidase activity and altered cell morphology. The protein expression of the senescence marker prelamin A was
also determined.
RESULTS
1) Long-term treatment with RTV and LPV/r resulted in progressive endothelial cell dysfunction
2) Long-term treatment with RTV and LPV/r progressively induced cellular senescence
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