Parthenolide alleviates doxorubicin-induced cardiotoxicity via regulation of Cyp1a1 and Nppa and suppression of NLRP3 inflammasome

Figures

• 

  Fig. 1. PTL attenuated Dox-induced cardiac dysfunction in mice. (A) The chemical structure of PTL. (B) Schematic diagram of the animal dosage regimen. (C) Representative M-mode echocardiographs of left ventricular chamber and (D–G) the calculation of FS%, EF%, LVIDs and LVIDd. (H–K) Measurement of myocardial injury biomarkers including LDH (H), CK (I), CK-MB (J), and cTnT (K). Con: control; Dox: doxorubicin; PTL: Parthenolide; PTL-10: 10 mg/kg; PTL-20: 20 mg/kg; EF%: left ventricular ejection fraction; FS%: left ventricular fractional shortening; LVIDs: left ventricular end-systolic internal dimension; LVIDd: left ventricular end-diastolic internal; LDH: lactate dehydrogenase; CK: creatinine kinase; CK-MB: creatinine kinase-MB; cTnT: troponin T. Data are mean ± S.E.M., n = 6–8, ∗P < 0.05, ∗∗P < 0.01, vs Con; #P < 0.05, ##P < 0.01, vs Dox.

• 

  Fig. 2. PTL suppressed Dox-induced cardiomyocyte apoptosis in mice. (A) Representative images of H&E staining in the myocardium. (B) Representative images of TUNEL staining. (C–D) Protein level of the Bcl-2/Bax ratio in cardiac tissues of mice, as measured by western blotting. Con: control; Dox: doxorubicin; PTL: parthenolide; PTL-10: 10 mg/kg; PTL-20: 20 mg/kg. Data are mean ± S.E.M., n = 6, ∗P < 0.05, ∗∗P < 0.01, vs Con; #P < 0.05, ##P < 0.01, vs Dox.

• 

  Fig. 3. PTL attenuated Dox-induced cardiomyocyte apoptosis in H9c2 cells. (A) Cell viability of H9c2 cells pretreated with PTL (0.5–5 μM) for 1 h and then treated with both Dox and PTL for 48 h. (B–C) Apoptosis rate was determined by flow cytometry for H9c2 cells pretreated with PTL for 1 h and then treated with Dox and PTL for 48 h. (D–E) Protein level of the Bcl-2/Bax ratio in H9c2 cells. Con: control; Dox: doxorubicin; PTL: parthenolide; PTL-0.5: 0.5 μM; PTL-1: 1 μM. Data are mean ± S.E.M., n = 5. ∗P < 0.05, ∗∗P < 0.01 vs. Con; #P < 0.05, ##P < 0.01 vs. Dox.

• 

  Fig. 4. The analysis of RNA-seq. (A) Correlation heat map between pairs based on expression abundance. The redder the color, the higher the correlation is. (B) PCA diagram. (C) The Venn map of DEGs shows 157 genes downregulated by Dox and upregulated by PTL, and 110 genes upregulated by Dox and downregulated by PTL. (D) Cluster heat map of the top 100 DEGs with the largest |log2FC| values. A change in color from blue to red notes the expression level of the gene from low to high. (E–F) GO (E) and KEGG (F) analysis of the top 100 DEGs. The data for each group were obtained from six mice.

• 

  Fig. 5. RT-qPCR verification for RNA-seq. (A–F) Quantitative mRNA expression of Cyp1a1, Nppa, Rnf144b, Timp4, Ucp3 and Pdk4 in RNA-seq analysis. (G–L) The mRNA level of Cyp1a1, Nppa, Rnf144b, Timp4, Ucp3 and Pdk4 detected by RT-qPCR. Con: control; Dox: doxorubicin; PTL: parthenolide; PTL-20: 20 mg/kg. Data are mean ± S.E.M., n = 6–8, ∗P < 0.05, ∗∗P < 0.01, vs Con; #P < 0.05, ##P < 0.01, vs Dox.

• 

  Fig. 6. PTL can inhibit NLRP3 inflammasome-mediated inflammation in Dox-treated mice. (A–B) The IL-1β (A) and NLRP3 (B) immunohistochemistry in cardiac tissues. (C–D) The representative results of NLRP3, Casepase 1, pro IL-1β and cleaved IL-1β expression in cardiac tissues calculated by western blotting. (E–F) Quantification of results from (C–D). (H) The representative results of NLRP3, pro IL-1β and cleaved IL-1β expression in H9c2 cells measured by western blotting. (I–J) Quantification of results from (H). Con: control; Dox: doxorubicin; PTL: parthenolide; PTL-10: 10 mg/kg; PTL-20: 20 mg/kg; PTL-0.5: 0.5 μM; PTL-1: 1 μM. Data are mean ± S.E.M., n = 6–8 (mice) or n = 5 (H9c2 cell), ∗P < 0.05, ∗∗P < 0.01 vs. Con; #P < 0.05, ##P < 0.01 vs. Dox.

• 

  figs1.

• 

  figs2.

• 

  figs3.

• 

  figs4.

• 

  figs5.

• 

  figs6.