AIM: To explore the mechanisms involved in ox-LDL transcytosis across endothelial cells and the role of caveolae in this process. METHODS: An in vitro model was established to investigate the passage of oxidized low density lipoprotein (ox-LDL) through a tight monolayer of human umbilical vein endothelial cells (HUVEC) cultured on a collagen-coated filter. Passage of DiI-labeled ox-LDL through the monolayer was measured using a fluorescence spectrophotometer. The uptake and efflux of ox-LDL by HUVEC were determined using fluorescence microscopy and HPLC. RESULTS: Caveolae inhibitors - carrageenan (250 μg/mL), filipin (5 μg/mL), and nocodazole (33 μmol/L)-decreased the transport of ox-LDL across the monolayer by 48.9%, 72.4%, and 79.8% as compared to the control group. In addition, they effectively decreased ox-LDL uptake and inhibited the efflux of ox-LDL. Caveolin-1 and LOX-1 were up-regulated by ox-LDL in a time-dependent manner and decreased gradually after depletion of ox-LDL (P<0.05). After treatment HUVEC with ox-LDL and silencing caveolin-1, NF-κB translocation to the nucleus was blocked and LOX-1 expression decreased (P<0.05). CONCLUSION: Caveolae can be a carrier for ox-LDL and may be involved in the uptake and transcytosis of ox-LDL by HUVEC.
AIM: To explore the mechanisms involved in ox-LDL transcytosis across endothelial cells and the role of caveolae in this process. METHODS: An in vitro model was established to investigate the passage of oxidized low density lipoprotein (ox-LDL) through a tight monolayer of human umbilical vein endothelial cells (HUVEC) cultured on a collagen-coated filter. Passage of DiI-labeled ox-LDL through the monolayer was measured using a fluorescence spectrophotometer. The uptake and efflux of ox-LDL by HUVEC were determined using fluorescence microscopy and HPLC. RESULTS: Caveolae inhibitors - carrageenan (250 μg/mL), filipin (5 μg/mL), and nocodazole (33 μmol/L)-decreased the transport of ox-LDL across the monolayer by 48.9%, 72.4%, and 79.8% as compared to the control group. In addition, they effectively decreased ox-LDL uptake and inhibited the efflux of ox-LDL. Caveolin-1 and LOX-1 were up-regulated by ox-LDL in a time-dependent manner and decreased gradually after depletion of ox-LDL (P<0.05). After treatment HUVEC with ox-LDL and silencing caveolin-1, NF-κB translocation to the nucleus was blocked and LOX-1 expression decreased (P<0.05). CONCLUSION: Caveolae can be a carrier for ox-LDL and may be involved in the uptake and transcytosis of ox-LDL by HUVEC.
Authors: Philippe G Frank; Scott E Woodman; David S Park; Michael P Lisanti Journal: Arterioscler Thromb Vasc Biol Date: 2003-04-10 Impact factor: 8.311
Authors: Dorothy I Mundy; Thomas Machleidt; Yun-shu Ying; Richard G W Anderson; George S Bloom Journal: J Cell Sci Date: 2002-11-15 Impact factor: 5.285
Authors: Ayesha N Shajahan; Barbara K Timblin; Raudel Sandoval; Chinnaswamy Tiruppathi; Asrar B Malik; Richard D Minshall Journal: J Biol Chem Date: 2004-03-08 Impact factor: 5.157
Authors: Chinnaswamy Tiruppathi; Tabassum Naqvi; Yubin Wu; Stephen M Vogel; Richard D Minshall; Asrar B Malik Journal: Proc Natl Acad Sci U S A Date: 2004-05-10 Impact factor: 11.205
Authors: Gregory M Lanza; Chrit Moonen; James R Baker; Esther Chang; Zheng Cheng; Piotr Grodzinski; Katherine Ferrara; Kullervo Hynynen; Gary Kelloff; Yong-Eun Koo Lee; Anil K Patri; David Sept; Jan E Schnitzer; Bradford J Wood; Miqin Zhang; Gang Zheng; Keyvan Farahani Journal: Wiley Interdiscip Rev Nanomed Nanobiotechnol Date: 2013-10-31
Authors: Susan M Armstrong; Michael G Sugiyama; Karen Y Y Fung; Yizhuo Gao; Changsen Wang; Andrew S Levy; Paymon Azizi; Mark Roufaiel; Su-Ning Zhu; Dante Neculai; Charles Yin; Steffen-Sebastian Bolz; Nabil G Seidah; Myron I Cybulsky; Bryan Heit; Warren L Lee Journal: Cardiovasc Res Date: 2015-09-02 Impact factor: 10.787