Bay 11-7085

BAY 11-7085 inhibits TNFα-induced expression of adhesion molecules E-selectin, VCAM-1, and ICAM-1 by inhibition of NF-κB without detectable cytotoxicity at 10 μM in HUVEC cells. ^[1] BAY 11-7085 enhances the inhibition of NFkappaB activity and decreases the viability of cells. ^[3] Besides, combination of Bay11-7085 and LY294002 leads to synergistic apoptotic effect in PEL cells. ^[4]

BAY 11-7085 inhibits the meningitis-associated increase in NF-κB activity, and results in an improvement of the clinical status of infected rats and a marked attenuation of meningitis-associated CNS complications and meningeal inflammation. ^[2] BAY 11-7085 increases the efficacy of induced inhibition of intraabdominal dissemination and production of ascites in athymic nude mice bearing Caov-3 cells. ^[3]

• In Gel Kinase Assay

  In gel kinase assay for the proteins that phosphorylate IκB-α is carried out as detailed below. Whole cell extracts are prepared from HUVEC treated with TNFα (100 units/ml) for 15 min in the presence or absence of inhibitor (20 μM, pretreatment for 1 h) as indicated. Proteins are separated on a 10% SDS gel containing 0.5 mg/ml HIS-IκB-α. Gels are washed two times in 20% propanol, 50 mM Hepes, pH 7.6, for 30 min and two times in buffer A (50 mM Hepes, pH 7.6, 5 mM 2-mercaptoethanol) for 30 min, followed by a 1-h incubation with buffer A, 0.05% Tween 20 and 1 h in buffer A with 0.05% Tween 20. The kinase assay is carried out for 1 h at 30 °C in the presence of 50 μM ATP, 5 μCi/ml [³²P]ATP, 20 mM Hepes, pH 7.6, 20 mM MgCl₂, 20 mM β-glycerophosphate, 20 mM p-nitrophenyl phosphate, 1 mM sodium vanadate, 2 mM dithiothreitol. The gel is washed with 5% trichloroacetic acid and 1% sodium pyrophosphate, dried, and exposed to film. A separate gel with no HIS-IκB-α is assayed as a control.

• Cell lines

  HUVEC cells

• Concentrations

  ~20 μM

• Incubation Time

  16 hours

• Method

  Human umbilical vein endothelial cells (HUVEC) are isolated and maintained in culture. Cell toxicity is assessed by morphology and by MTT assay.

• Animal Models

  Rat model of pneumococcal meningitis

• Dosages

  PBS

• Administration

  i.p.