Pirfenidone and Vitamin D Ameliorate Cardiac Fibrosis Induced by Doxorubicin in Ehrlich Ascites Carcinoma Bearing Mice: Modulation of Monocyte Chemoattractant Protein-1 and Jun N-terminal Kinase-1 Pathways

Figures

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  Figure 1. Effect of individual treatment with doxorubicin (15 mg/kg; i.p.), pirfenidone (500 mg/kg; p.o.) or vitamin D (0.5 µg/kg; i.p.) and their combinations on the levels of biochemical markers (A) serum troponin and (B) creatine kinase (CK) (C) CK-MB in EAC bearing mice on day 21. Results are expressed as mean ± SEM. All data were analyzed using ANOVA followed by Bonferroni’s post-hoc test at p < 0.05. & p < 0.05 with respect to normal, * p < 0.05 with respect to control EAC, # p < 0.05 with respect to doxorubicin. $ p < 0.05 with respect to the co-therapy of doxorubicin, pirfenidone, and vitamin D (n = 5).

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  Figure 2. Effect of individual treatment with doxorubicin (15 mg/kg; i.p.), pirfenidone (500 mg/kg; p.o.) or vitamin D (0.5 µg/kg; i.p.) and their combinations on cardiac expression of NF-κB, and MCP-1 in EAC bearing mice on day 21. The bar chart demonstrates NF-κB. (A) A graph showing the density of immunostaining for NF-κB in experimental groups. (B) Photomicrographs showing cardiac expression of NF-κB. (C) RT-PCR measurement of MCP-1 in cardiac tissue. & p < 0.05 with respect to normal, * p < 0.05 with respect to control EAC, # p < 0.05 with respect to doxorubicin. $ p < 0.05 with respect to the co-therapy of doxorubicin, pirfenidone, and vitamin D (n = 5).

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  Figure 3. Effect of individual treatment with doxorubicin (15 mg/kg; i.p.), pirfenidone (500 mg/kg; p.o.) or vitamin D (0.5 µg/kg; i.p.) and their combinations on cardiac smad3, smad7 and JNK1 expressions in EAC bearing mice. (A) A graph showing quantitative analysis of the relative expression of different proteins after treatment. (B) Western blot analysis showing protein expression of cardiac smad3, smad7, and JNK1 and β-actin in different experimental groups. (C) RT-PCR measurement of TGF-β1 in cardiac tissue. & p < 0.05 with respect to normal, * p < 0.05 with respect to control EAC, # p < 0.05 with respect to doxorubicin. $ p < 0.05 with respect to the co-therapy of doxorubicin, pirfenidone, and vitamin D (n = 5).

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  Figure 4. Effect of individual treatment with doxorubicin (15 mg/kg; i.p.), pirfenidone (500 mg/kg; p.o.) or vitamin D (0.5 µg/kg; i.p.) and their combinations on the cardiac expression of α-SMA in EAC bearing mice on day 21. (A) A graph showing the density of immunostaining for α-SMA in experimental groups. (B) Photomicrographs showing cardiac expression of α-SMA. & p < 0.05 with respect to normal, * p < 0.05 with respect to control EAC, # p < 0.05 with respect to doxorubicin. $ p < 0.05 with respect to the co-therapy of doxorubicin, pirfenidone, and vitamin D (n = 5).

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  Figure 5. Representative photomicrographs for sections from the heart showing the effect of individual treatment with doxorubicin (15 mg/kg; i.p.), pirfenidone (500 mg/kg; p.o.) or vitamin D (0.5 µg/kg; i.p.) and their combinations on cardiac tissues fibrosis assessed by Masson trichrome staining in EAC bearing mice on day 21.