Paeoniflorin improves myocardial injury via p38 MAPK/NF-KB p65 inhibition in lipopolysaccharide-induced mouse

Figure 1. The effects of Pae on cardiac injury in mice. LPS treatment 12 h later, cardiac cell infiltration was detected by HE staining (A), images were magnified at 200×, and the level of cTnI (B), LDH (C) and CK-MB (D) was detected by ELISA. Data are expressed as the mean ± SD (n=8). *, P<0.05 versus the sham group; #, P<0.05 versus the LPS group. LPS, lipopolysaccharide; Pae, paeoniflorin; cTnI, cardiac troponin I; LDH, lactate dehydrogenase; CK-MB, creatine kinase-MB.

Figure 2. The effects of Pae on inflammatory response. (A) The levels of IL-6, TNF-α, and IL-1β in myocardial tissues were measured by ELISA assay. (B) The levels of IL-6, TNF-α, and IL-1β in peripheral blood were measured by ELISA assay. Data are expressed as the mean ± SD (n=8). *, P<0.05 versus the sham group; #, P<0.05 versus the LPS group. LPS, lipopolysaccharide; Pae, paeoniflorin; TNF-α, tumor necrosis factor-α; IL, interleukin.

Figure 3. The effects of Pae on oxidative stress and mitochondrial injury. The levels of MDA (A), GSH (B), ROS (C) and SOD (D) were detected by ELISA assay. (E) The absorbance of mPTP was measured by an ultraviolet spectrophotometer. (F) JC-1 staining showed the change of MMP, images were obtained by a fluorescence microscope, Bar =75 µm. Data are expressed as the mean ± SD (n=8). *, P<0.05 versus the sham group; #, P<0.05 versus the LPS group. Pae, paeoniflorin; MDA, malondialdehyde; GSH, glutathione; ROS, reactive oxygen species; SOD, superoxide dismutase; MMP, mitochondrial membrane potential.

Figure 4. The effects of Pae on cardiac apoptosis. (A) The apoptosis of myocardial tissues was examined by TUNEL staining, images were magnified at 200×. (B) The protein expression of Cytochrome C and Caspase-3, Bax, and Bcl-2 were examined by western blotting. Data are expressed as the mean ± SD (n=8). *, P<0.05 versus the sham group; #, P<0.05 versus the LPS group. LPS, lipopolysaccharide; Pae, paeoniflorin.

Figure 5. The effects of Pae on activity of p38 MAPK and NF-κB p65. (A) The protein expression of p38, p-p38, p65 and p-p65 were measured by western blotting. After adding p38 inhibitor SB203580, (B) cardiac cell infiltration was detected by HE staining, images were magnified at 200×. (C,D) The apoptosis of myocardial tissues was examined by TUNEL staining, representative images were magnified at 200×. The levels of TNF-α (E), and IL-1β (F) in myocardial tissues were measured using ELISA assay. The serum levels of MDA (G) and SOD (H) were detected using ELISA assay. Data are expressed as the mean ± SD (n=8). *, P<0.05 versus the sham group; #, P<0.05 versus the LPS group; &, P<0.05 versus the SB203580 or Pae group. Pae, paeoniflorin; TNF-α, tumor necrosis factor-α; IL, interleukin; MDA, malondialdehyde; SOD, superoxide dismutase.