Efficacy of AdipoDren® in Reducing Interleukin-1-Induced Lymphatic Endothelial Hyperpermeability

Abstract

Lymphatic leakage can be seen as a detrimental phenomenon associated with fluid retention and deposition as well as gain of weight. Moreover, lymphatic dysfunction is associated with an inflammatory environment and can be a substrate for other health conditions. A number of treatments can ameliorate lymphatic vasculature: natural substances have been used as treatment options particularly suitable for their consolidated effectiveness and safety profile. Here we report the protective effect of AdipoDren®, an association of a series of plant-derived natural complexes, on lymphatic endothelium permeability promoted by interleukin-1 beta (IL-1β) and the associated molecular mechanisms. AdipoDren® demonstrated a protective effect on dermal lymphatic endothelial cell permeability increased by IL-1β. Reduced permeability was due to the maintenance of tight junctions and cell-cell localisation of occludin and zonula occludens-1 (ZO-1). Moreover, AdipoDren® reduced the expression of the inflammatory key element cyclooxygenase-2 (COX-2), while not altering the levels of endothelial and inducible nitric oxide synthases (eNOS and iNOS). The upregulation of antioxidant enzymatic systems (catalase and superoxide dismutase-1, SOD-1) and the downregulation of pro-oxidant markers (p22 phox subunit of NADPH oxidase) were also evident. In conclusion, AdipoDren® would be useful to ameliorate conditions of altered lymphatic vasculature and to support the physiological functionality of the lymphatic endothelium.

ALDH2 Activity Reduces Mitochondrial Oxygen Reserve Capacity in Endothelial Cells and Induces Senescence Properties

“…Cells were seeded (8 × 10 4 cells/well) 24 h post-ALDH2 silencing on gelatin-coated insert membranes (0.4 μ m diameter pores, Corning, New York, USA), and the inserts were placed in 24-multiwell plates and incubated for 24 h. Next, cells were exposed to EBM-2 with 0.1% FBS for 8 h. The assay was carried out as described [ 20 ]. Briefly, the fluorescent permeability tracer (3 kDa FITC-Dextran, 10 μ M) was added, and the fluorescence was measured after 7 min in the medium present in the bottom of the well, in a multiplate reader (Infinite 200 Pro, SpectraFluor, Tecan), at 485/535 nm (excitation/emission).…”

“…Briefly, the fluorescent permeability tracer (3 kDa FITC-Dextran, 10 μ M) was added, and the fluorescence was measured after 7 min in the medium present in the bottom of the well, in a multiplate reader (Infinite 200 Pro, SpectraFluor, Tecan), at 485/535 nm (excitation/emission). Results are reported as relative fluorescence units (RFU) [ 20 ].…”

“…siCTR and siALDH2 cells (24 h posttransfection) were seeded (1.5 × 10 3 cells/well) in triplicate in a 96-well plate, and after adherence (6–8 h), the medium was replaced with EBM-2 with 0.1% FBS in the presence or absence of 5 mM NAC for 24 h. DCFH2-DA (2,-7-dichlorodihydrofluorescein diacetate, Invitrogen) was used, and intracellular ROS were measured as previously described [ 20 ]. The results are reported as fold change vs. siCTR of relative fluorescence units (RFU) corrected for the cell number counted [ 20 ].…”

Pharmacological Inhibition of CA-IX Impairs Tumor Cell Proliferation, Migration and Invasiveness

“…Protein expression was evaluated by western blot [48]. In total, 3 × 10 5 cells were plated in 60 mm dishes, with complete DMEM supplemented with 10% FBS.…”

The metal-nonoate Ni(SalPipNONO) inhibitsin vitrotumor growth, invasiveness and angiogenesis

“…ROS levels were evaluated as previously reported [ 60 ]. A total of 1.5 × 10 3 cells were seeded in 96-well plates and, after adherence, were treated with 0.5 mM Ni(SalPipNONO) or Ni(SalPip) in medium without phenol red and different serum concentrations.…”