Glaucocalyxin A Attenuates Allergic Responses by Inhibiting Mast Cell Degranulation through p38MAPK/NrF2/HO-1 and HMGB1/TLR4/NF-κB Signaling Pathways

Figure 5

Effect of GLA on HMGB1-mediated signaling pathways in RBL-2H3 cells. Protein levels of HMGB1/TLR4/NF-κB signaling pathway were measure by Western blot. (a) and (b) RBL-2H3 cells were challenged with 100 ng/ml DNP-HSA in the presence or absence of GLA. Protein levels were analyzed. (c) RBL-2H3 cells were stimulated with 20 ng/ml rmHMGB1 with or without GLA (5.0 μM) for 30 min. Protein levels were analyzed. (d) The effect of GLA on proinflammatory cytokines in the rmHMGB1-stimulated RBL-2H3 cells. (e) Release of β-hexosaminidase. (f) Calcium uptake was measured by the radioenzymatic method. (g) RBL-2H3 cells were transfected with 50 nM control (siCont) or HMGB1 siRNA (si-HMGB1) for 6 h. After 24 h, they were then stimulated with DNP HSA (100 ng/mL). (h) The release of β-hexosaminidase was measured after cell treatment. (i) Calcium uptake was measured after cell treatment. Protein levels were analyzed. Each datum represents the mean ± SEM (n = 3). Compared with the IgE + Ag group or rmHMGB1 group, . Compared with IgE + Ag + GLA group, .

(a)

(b)

(c)

(d)

(e)

(f)

(g)

(h)

(i)