Purified low-density lipoprotein and bovine serum albumin efficiency to internalise lycopene into adipocytes.

Epidemiological studies have suggested that lycopene has protective effects against various diseases including cardiovascular diseases. However, mechanistic studies to understand these effects are difficult due to the insolubility of lycopene in aqueous culture medium. The objective of the present study was to use LDL or BSA as physiological vehicles for lycopene and to compare them with various classical vehicles. Among tested vehicles, only LDL, BSA, THF/BHT, beadlets, and liposomes were able to solubilise lycopene. No cytotoxicity was observed with these vehicles. LDL and BSA allowed good stability of lycopene during incubation (52% and 43% for 2microM lycopene solutions), but remained less efficient than THF/BHT or beadlets (67% and 62%). Incubation of adipocytes (3T3-L1) with the different vehicles for 24 and 48h showed that beadlets best delivered lycopene to cells. Finally, whatever the vehicle used, intracellular localization of lycopene was the same: lipid droplets (32-51%), plasma membrane (32-37%) and nuclear membrane (19-29%). As a conclusion, LDL or BSA display comparable properties to THF/BHT or beadlets. It is the first time that lycopene carried by physiological vehicles is shown to reach different subcellular compartments supporting molecular effects in adipocyte, such as cell signaling or nuclear receptor interacting.

The plasminogen activation system modulates differently adipogenesis and myogenesis of embryonic stem cells.

Regulation of the extracellular matrix (ECM) plays an important functional role either in physiological or pathological conditions. The plasminogen activation (PA) system, comprising the uPA and tPA proteases and their inhibitor PAI-1, is one of the main suppliers of extracellular proteolytic activity contributing to tissue remodeling. Although its function in development is well documented, its precise role in mouse embryonic stem cell (ESC) differentiation in vitro is unknown. We found that the PA system components are expressed at very low levels in undifferentiated ESCs and that upon differentiation uPA activity is detected mainly transiently, whereas tPA activity and PAI-1 protein are maximum in well differentiated cells. Adipocyte formation by ESCs is inhibited by amiloride treatment, a specific uPA inhibitor. Likewise, ESCs expressing ectopic PAI-1 under the control of an inducible expression system display reduced adipogenic capacities after induction of the gene. Furthermore, the adipogenic differentiation capacities of PAI-1(-/-) induced pluripotent stem cells (iPSCs) are augmented as compared to wt iPSCs. Our results demonstrate that the control of ESC adipogenesis by the PA system correspond to different successive steps from undifferentiated to well differentiated ESCs. Similarly, skeletal myogenesis is decreased by uPA inhibition or PAI-1 overexpression during the terminal step of differentiation. However, interfering with uPA during days 0 to 3 of the differentiation process augments ESC myotube formation. Neither neurogenesis, cardiomyogenesis, endothelial cell nor smooth muscle formation are affected by amiloride or PAI-1 induction. Our results show that the PA system is capable to specifically modulate adipogenesis and skeletal myogenesis of ESCs by successive different molecular mechanisms.

CD36 is involved in lycopene and lutein uptake by adipocytes and adipose tissue cultures.

SCOPE: Carotenoids are mainly stored in adipose tissue. However, nothing is known regarding the uptake of carotenoids by adipocytes. Thus, our study explored the mechanism by which lycopene and lutein, two major human plasma carotenoids, are transported. METHODS AND RESULTS: CD36 was a putative candidate for this uptake, 3T3-L1 cells were treated with sulfosuccinimidyl oleate, a CD36-specific inhibitor. sulfosuccinimidyl oleate-treated cells showed a significant decrease in both lycopene and lutein uptake as compared to control cells. Their uptake was also decreased by partial inhibition of CD36 expression using siRNA, whereas the overexpression of CD36 in Cos-1 cells increased their uptake. Finally, the effect of CD36 on carotenoid uptake was confirmed ex vivo in cultures of adipose tissue explants from CD36(-/-) mice, which exhibited reduced carotenoid uptake as compared to wild-type mice explants. CONCLUSION: For the first time, we report the involvement of a transporter, CD36, in carotenoid uptake by adipocytes and adipose tissue.