Fig. 6
Baicalin blocks the JNK signaling pathway and alleviates COPD in a HSP72 dependent manner. a MLE-12 cells were transfected with si-HSP72 or control vector and stimulated with CSE. The cells treated with baicalin and the phosphorylation of JNK normalized to GAPDH was analyzed by immunoblotting; *p < 0.05, **p < 0.01, ***p < 0.001 compared to untreated cells; #p < 0.05, ##p < 0.01, ###p < 0.001 compared to cells treated with CSE; &p < 0.05, ##p < 0.01, ###p < 0.001 compared to cells treated with CSE + baicalin. b The extent of JNK phosphorylation after Anisomycin treatment normalized to GAPDH was tested by immunoblotting. *p < 0.05, **p < 0.01, ***p < 0.001 compared to cells treated with vehicle. c The phosphorylation of JNK normalized to GAPDH was analyzed by immunoblotting. d Cell viability was analyzed by CCK-8. e Cell apoptosis was analyzed by flow cytometry. f Release of IL-6, IL-8 and TNF-α was analyzed ELISA. *p < 0.05, **p < 0.01, ***p < 0.001 compared to cells treated with CSE; #p < 0.05, ##p < 0.01, ###p < 0.001 compared to cells treated with CSE + baicalin. g Relative protein expression of HSP72 normalized to GAPDH after treatment of HSP72 overexpression vector was determined by immunoblotting. *p < 0.05, **p < 0.01, ***p < 0.001 compared to cells transfected with oe-NC. h Cells were co-transfected with both Anisomycin and oe-HSP72 and stimulated with CSE. The expression of HSP72 and extent of JNK phosphorylation normalized to GAPDH was analyzed by immunoblotting. i Cell viability of both JNK-activated and HSP72-overexpressed cells after treatment with baicalin was analyzed by CCK-8 assay. j Cell apoptosis of both JNK-activated and HSP72-overexpressed cells after treatment with baicalin was analyzed by flow cytometry. k Release of IL-6, IL-8 and TNF-α of both JNK-activated and HSP72-overexpressed cells after treatment with baicalin was analyzed ELISA. *p < 0.05, **p < 0.01, ***p < 0.001 compared to cells treated with CSE + baicalin + Anisomycin. All experiments were repeated for three times independently and the data were represented as mean ± standard deviation. One-way ANOVA followed by Tukey’s test was used for multiple group comparison