Fig. 8

NF-κB inhibitors impair HMGB1-induced platelet activation responses and elevated cGMP level. a, b NF-κB inhibitors impair platelet aggregation induced by HMGB1, WPs were treated with the indicated concentrations of BAY11-7082 in an aggregometer (1200 rpm) at 37 °C for 5 min and then stimulated with HMGB1 (n = 5, *P < 0.01 vs ALB). c HMGB1 induce IκBα phosphorylation and degradation, WPs were treated with various dose of HMGB1 for 5 min 37 °C, then Lysates were immunoblotted with an anti- IκBα Ab. d WPs preincubation with BAY 11-7082 under high shear (1200 rpm) in an aggregometer for 5 min and then stimulated with HMGB1 (40 μg/ml) for 5 min, HMGB1 induce IκBα phosphorylation can be blocked by BAY 11-7082. (n = 4, P < 0.01 vs control). e Effects BAY 11-7082 on HMGB1-induced cAMP and cGMP, WPs pretreatment with BAY11-7082 in an aggregometer and stimulated with HMGB1 (40 μg/ml) for 5 min before termination of the reaction and then cAMP and cGMP enzyme immunoassays were performed. BAY11-7082 significantly increased cGMP accumulation in a dose dependent manner. (n = 4, *P < 0.01 vs. control). f Effects of a-TLR4, SNP, BAY 11-7082 or ODQ on HMGB1-induced IκBα phosphorylation. WPs were stirred in an aggregometer with above materials for 5 min prior to the addition of HMGB1 (40 μg/ml). a-TLR4 (50 μg/ml), SNP (100 μM) ODQ (20 μM) and BAY 11-7082 (40 μM), measured IκBα phosphorylation level, all immunoblots are representative of 3 to 4 similar experiments