Figure 7

IVIg protects primary neurons against HMGB1-induced cell death by modulating TLRs expression. (A, B) Different concentrations of mouse recombinant HMGB1 significantly increase oxygen and glucose deprivation (OGD)-induced pro-apoptotic cleaved caspase-3 level. Data are represented as mean ± SEM. n = 3 cultures. ⁺ P < 0.05 in comparison with OGD 4.5-h group; ⁺⁺ P < 0.01 in comparison with OGD 4.5-h group; ns, not significant. (C-E) Representative immunoblots and quantification illustrating HMGB1 increases the levels of TLR2 and TLR4 in primary cortical neurons following 4.5 h of OGD. Administration of IVIg (10 mg/ml) significantly prevents HMGB1-induced increased expression level of TLR2 and TLR4. Data are represented as mean ± SEM. n = 5 cultures. ⁺ P < 0.05 in comparison with OGD 4.5-h group; ⁺⁺ P < 0.01 in comparison with OGD 4.5-h group; *P < 0.05 in comparison with corresponding OGD group; **P < 0.01 in comparison with corresponding OGD group; ***P < 0.001 in comparison with corresponding OGD group. (F-H) Representative immunoblots and quantification illustrating HMGB1 increases the levels of MyD88 and TRAF6 in primary cortical neurons following 4.5 h of OGD. Administration of IVIg (10 mg/ml) significantly prevents HMGB1-induced increased expression level of MyD88 and TRAF6. Data are represented as mean ± SEM. n = 5 cultures. ⁺ P < 0.05 in comparison with OGD 4.5-h group; *P < 0.05 in comparison with corresponding OGD group; ***P < 0.001 in comparison with corresponding OGD group.