Repurposing lurasidone to alleviate doxorubicin-induced cardiotoxicity and neurotoxicity via BDNF/TrkB/PI3K/Akt/CREB and miR-34a-5p/PGC-1α pathways

Figures

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  Fig. 1. The experimental design of the work

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  Fig. 2. Venn diagram of the intersecting targets for lurasidone and doxorubicin-induced toxicity (DIT)

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  Fig. 3. Network analysis of common targets, as generated by the STRING database. A The core PPI network of the 54 common target proteins, comprising 54 nodes and 150 edges. B The expanded PPI network after the integration of 100 top first- and second-shell interactors, resulting in a comprehensive network of 154 nodes and 4309 edges. C PPI network of the twelve experimentally validated targets generated

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  Fig. 4. Integrated drug-disease-target-pathway network, ellipse: drug, rectangle: disease, diamond: targets, V-shape: pathways

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  Fig. 5. Top 20 significantly enriched analyses. A Biological process. B Cellular component. C Molecular function. D KEGG pathways. All are sorted by fold enrichment

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  Fig. 6. Effect of Lura (1 and 3 mg/kg) on behavioralchanges in Dox-treated rats. A Number of grooming in OFT. B Number of rearing in OFT. C Ambulation frequency in an OFT. D Time of immobility in FST. The outcomes have been reported by way of mean ± SD (n = 6). One-way ANOVA was used to test significance, ensued by Tukey–Kramer multiple comparisons. *: conveys statistical disparity from the control group; #: conveys statistical disparity from the Dox group; $: conveys statistical disparity from the Lura 1 mg + Dox group, at p < 0.05. Dox: doxurubicin; Lura: lurasidone; FST: forced swimming test; OFT: open field test

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  Fig. 7. Effect of Lura (1 and 3 mg/kg) on neuronal and cardiac biomarkers in Dox-treated rats. Neuronal markers. A AchE. B DA; cardiac markers. C cTn-I. D CK-MB. The outcomes are reported as mean ± SD (n = 6). One-way ANOVA was used to test significance, followed by Tukey–Kramer multiple comparisons. *: conveys statistical disparity from the control group; #: conveys statistical disparity from the Dox group; $: conveys statistical disparity from the Lura 1 mg + Dox group at p < 0.05. Dox: doxorubicin; Lura: lurasidone; AchE: acetylcholinesterase; DA: dopamine; cTn-I: cardiac troponin-I; CK-MB: creatinine kinase-MB

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  Fig. 8. Effect of Lura (1 and 3 mg/kg) on neuronal and cardiac oxidative stress markers in Dox-treated rats. Neuronal markers. A GSH content. B SOD activity, cardiac markers. C GSH content. D SOD activity. The outcomes have been reported by way of mean ± SD (n = 6). One-way ANOVA was used to test significance, ensued by Tukey–Kramer multiple comparisons. *: conveys statistical disparity from the control group; #: conveys statistical disparity from the Dox group; $: conveys statistical disparity from the Lura 1 mg + Dox group, at p < 0.05. Dox: doxorubicin; Lura: lurasidone; GSH: glutathione; SOD: superoxide dismutase

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  Fig. 9. Effect of Lura (1 and 3 mg/kg) on neuronal and cardiac inflammatory markers in Dox-treated rats. Neuronal markers: NF-κB (A), TNF-α (B), and IL-1β (D); cardiac markers: NF-κB (D), TNF-α (E), and IL-1β (F) levels. G, I Representative photomicrography displays expression of neuronal GFAP and Iba- in the hippocampal region (CA3), respectively, in different groups. H, J Area % of GFAP expression and number of Iba-1 positive cells in the hippocampus (CA3 region), respectively (scale bars = 50 μm). K, M p50 subunit of NF-ĸB expression in neuronal and cardiac tissue, respectively. L, N Representative western blot images of p50 subunit of NF-ĸB expression in neuronal and cardiac tissue, respectively. The outcomes have been reported by way of mean ± SD (n = 6). One-way ANOVA was used to check significance, followed by Tukey–Kramer multiple comparisons. *: conveys statistical disparity from the oversight group; #: conveys statistical disparity from the Dox group; $: conveys statistical disparity from the Lura 1 mg + Dox group, at p < 0.05. Dox: doxorubicin; Lura: lurasidone; NF-κB: nuclear factor kappa B; TNF-α: tumor necrosis factor-alpha; IL-1β: interleukin-1 beta; Iba-1: ionized…

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  Fig. 10. Effect of Lura (1 and 3 mg/kg) on neuronal and cardiac Casp 3 levels in rats and on the number of intact neuron cells in Dox-treated rats. Neuronal Casp 3 content (A), cardiac Casp 3 content (B), number of intact neuron cells in the CA3 region (D), % of area of cleaved Casp 3 expression in the CA3 region (F), and % of area of cleaved Casp 3 expression in cardiac tissue (H). Representative photomicrographs of brain and heart sections displaying the number of intact neurons in the CA3 region after staining with toluidine blue (C), cleaved Casp 3 expression in the CA3 region (E), and cleaved Casp 3 expression in cardiac tissue (G) (scale bars = 50 μm). The outcomes have been reported by way of mean ± SD (n = 6). One-way ANOVA was applied to check significance, followed by Tukey–Kramer multiple comparisons. *: conveys statistical disparity from the control group; #: conveys statistical disparity from the Dox group; $: conveys statistical disparity from the Lura 1 mg + Dox group, at p < 0.05. Dox: doxorubicin; Lura: lurasidone; Casp 3: caspase 3

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  Fig. 11. The effect of Lura (1 and 3 mg/kg) on neuronal and cardiac expression of the BDNF, TrkB, PI3K, Akt, and p-CREB protein levels in Dox-treated rats. Neuronal expression of BDNF (A), TrkB (B), p-Akt (C), p-CREB (D), p-PI3k (L); cardiac expression of BDNF (F), TrkB (G), p-Akt (H), p-CREB (I), p-PI3k expression (N) levels. Representative western blot images of neuronal and cardiac contents of TrkB, BDNF, p-Akt, and p-CREB proteins, respectively, (E, J), and representative photomicrographs displaying hippocampal (CA3) and cardiac expression of p-PI3K, respectively (K, M). The outcomes have been reported by way of mean ± SD (n = 6). One-way ANOVA was employed to check significance, ensued by Tukey–Kramer multiple comparisons. *: conveys statistical disparity from the oversight group; #: conveys statistical disparity from the Dox group; $: conveys statistical disparity from the Lura 1 mg + Dox group, at p < 0.05. Dox: doxorubicin; Lura: lurasidone; BDNF: brain-derived neurotrophic factor; TrkB: tropomyosin receptor kinase B (TrkB); PI3K: phosphoinositide 3-kinase; p-Akt: phosphorylated protein kinase B (p-AKT); p-CREB: phosphorylated cAMP-response element binding protein; β actin:…

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  Fig. 12. Effect of Lura (1 and 3 mg/kg) on neuronal and cardiac mir-34a-5p and PCG-1α in Dox-treated rats. Neuronal: mir-34a-5p (A), PGC-1α (B) relative expression, cardiac; mir-34a-5p (C), PGC-1α (D) relative expression. The outcomes are reported as mean ± SD (n = 6). One-way ANOVA was used to assess significance, followed by Tukey–Kramer multiple comparisons. *: conveys statistical disparity from the control group; #: conveys statistical disparity from the Dox group; $: conveys statistical disparity from the Lura 1 mg + Dox group, at P < 0.05. Dox: doxorubicin; Lura: lurasidone; PCG-1α: peroxisome proliferator-activated receptor-gamma coactivator (PGC)−1alpha