Paeoniflorin Alleviates LPS‐Induced Inflammation and Acute Myocardial Injury by Inhibiting PI3K/Akt/ERK‐Mediated HIF‐1α and TLR4/MyD88/NF‐κB Inflammatory Signaling

Figure 1. PF improved cardiac dysfunction in SIMI rats. (A) Representative images of ECG recorded by lead II in different groups. (B) Representative images by echocardiograph in each group. (C–K) The statistical analysis of EF, FS, CO, LVPWs, LVPWd, LVAWs, LVAWd, LVIDs, and LVIDd in each group (n = 6). (L) The flowchart of the study. Significance: ⁣∗p < 0.05, ⁣∗∗p < 0.01 vs. the control group; #p < 0.05, ##p < 0.01 vs. the LPS group.

Figure 2. The PF-target-SIMI network construction. (A) Identification of intersection targets between PF and SIMI. (B) Construction of a “PF-SIMI-Intersection targets-signaling pathway” network.

Figure 3. PPI analysis of the targets of PF against SIMI. (A) The PPI network was obtained from STRING database. (B) The intersecting targets were visualized by Cytoscape. (C) The significant module was obtained from the PIN by MCODE method with a score >4.0; red represents the higher degree and yellow represents the lower degree.

Figure 4. GO and KEGG enrichment analysis of target genes of PF against SIMI. (A) Enriched GO terms for biological process (BP), cellular component (CC), and molecular function (MF) of potential anti-SIMI targets of PF. (B) Enriched KEGG pathways of potential anti-SIMI targets of PF.

Figure 5. Molecular docking of PF with PI3K, Akt1, ERK1, ERK2, HIF-1α, TLR4, and NF-κB.

Figure 6. PF mitigates LPS-induced acute myocardial injury. (A–E) The levels of serum LDH, CK-MB, cTnT/TNNT2, TNNI3/cTn-I, and AST. (F) HE staining of left ventricular tissue in each group. Scale bars = 100 μm or 50 μm. Significance: ⁣∗p < 0.05, ⁣∗∗p < 0.01 vs. the control group; #p < 0.05, ##p < 0.01 vs. the LPS group.

Figure 7. PF alleviates LPS-increased cardiac inflammatory response. (A, B) The levels of serum TNF-α and IL-1β. (C, D) The mRNA levels of TNF-α and IL-1β were detected by qRT-PCR. Significance: ⁣∗p < 0.05, ⁣∗∗p < 0.01 vs. the control group; #p < 0.05, ##p < 0.01 vs. the LPS group.

Figure 8. PF blocks TLR4/MyD88/NF-κB signaling in LPS-induced SIMI. (A) Western blot detection of TLR4, MyD88, P-NF-κB, and NF-κB. (B–F) Bar graphs show the relative expression of TLR4, Myd88, P-NF-κB, NF-κB, and the ratio of p-NF-κB/NF-κB as analyzed by western blot. β-actin was used as an internal control (n = 3). (G) Western blot detection of TLR4, P-NF-κB, and NF-κB. (H–K) Bar graphs show the relative expression of TLR4, p-NF-κB, NF-κB, and the ratio of p-NF-κB/NF-κB as analyzed by western blot. GAPDH was used as an internal control (n = 3). Data were shown as mean ± SEM. Significance: ⁣∗p < 0.05, ⁣∗∗p < 0.01 vs. the control group; #p < 0.05, ##p < 0.01 vs. the LPS group.

Figure 9. PF obstructs PI3K/Akt/ERK-mediated HIF-1α in LPS-induced SIMI. (A) Western blot detection of PI3K, P-Akt, Akt, P-ERK, ERK, and HIF-1α. (B–I) Bar graphs show the relative expression of PI3K, P-Akt, Akt, P-ERK, ERK, HIF-1α, and the ratio of P-Akt/Akt and P-ERK/ERK as analyzed by western blot. GAPDH was used as an internal control (n = 3). Data were shown as mean ± SEM. Significance: ⁣∗p < 0.05, ⁣∗∗p < 0.01 vs. the control group; #p < 0.05, ##p < 0.01 vs. the LPS group.

Figure 10. The schematic diagram for PF anti-SIMI by inhibiting PI3K/Akt/ERK-mediated HIF-1α and TLR4/MyD88/NF-κB.