Targeting SIRT-1/AMPK/Nrf2 Signaling Pathway by Tenofovir Protected Against Cyclophosphamide-Induced Nephrotoxicity and Cardiotoxicity in Rats

Figures

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  Figure 1. Experimental Design: Rats were randomly split up into five groups (n = 8) (i) The control group. (ii) cyclophosphamide (CYC) group. (iii) tenofovir 25 mg/kg group (TFV 25 mg/kg + CYC). (iv) tenofovir 50 mg/kg group (TFV 50 mg/kg + CYC). (v) tenofovir control (TFV 50 mg/kg) group. D: day, CMC: carboxymethyl cellulose, CYC: cyclophosphamide, I.P: intraperitoneal; TFV: tenofovir.

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  Figure 2. The impact of tenofovir (TFV) on serum lactate dehydrogenase (LDH), creatine kinase-myocardial band (CK-MB), serum creatinine (Cr), blood urea nitrogen (BUN), urine total protein (TP), and urine Cr altered via cyclophosphamide (CYC) injection. Data are expressed as mean ± standard error of the mean (SEM); n = 8. CYC (200 mg/kg, single i.p dose); TFV (25 or 50 mg/kg, oral for 7 days). The values of *, $, and # are significantly different from those of the control, CYC, and the (CYC + TFV 25 mg/kg) groups, respectively. This was determined using one-way ANOVA followed by Tukey–Kramer multiple comparison post-hoc tests (p < 0.05).

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  Figure 3. The impact of tenofovir (TFV) on renal histopathological alteration and damage scoring induced by cyclophosphamide (CYC) using hematoxylin and eosin stain. (A,a) Control group, (B,b) TFV control group, (C,c) CYC group, (D,d) CYC + TFV 25 group, (E,e) CYC + TFV 50 group. (F) Scatter dot plots displaying renal inflammation scores, (G) scatter dot plots displaying renal degeneration scores, (H) scatter dot plots displaying renal fibrosis scores, (I) scatter dot plots displaying renal vascular changes scores. Thin arrow: inflammation; thick arrow: interstitial fibrosis; arrowhead: intestinal congestion; twisted arrow: tubular degeneration or necrosis; circle: intraluminal cast; star: hemorrhage. Upper raw original magnification 10×, lower raw 40×, and scale bar 100 µm and 20 µm, respectively. CYC (200 mg/kg, single i.p dose); TFV (25 or 50 mg/kg, oral for 7 days). Data are presented as median ± IQR range; n = 4. The values of * were significantly different from those of the control group, as found with a Kruskal–Wallis test followed by a Dunn multiple comparison test (p ˂ 0.05).

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  Figure 4. The impact of tenofovir (TFV) on cardiac histopathological alteration and damage scoring induced by cyclophosphamide (CYC) using hematoxylin and eosin stain. (A,a) Control group, (B,b) TFV control group, (C,c) CYC group, (D,d) CYC + TFV 25 group, (E,e) CYC + TFV 50 group. (F) Scatter dot plots displaying cardiac inflammation scores, (G) scatter dot plots displaying cardiac degeneration scores, (H) scatter dot plots displaying cardiac congestion scores, (I) scatter dot plots displaying cardiac fibrosis scores. Twisted arrow: myocardial necrosis; thick arrow: fibrosis; thin arrow: inflammation; arrowhead: vascular congestion. Upper raw original magnification 10×, lower raw 40×, and scale bar 100 µm and 20 µm, respectively. CYC (200 mg/kg, single i.p dose); TFV (25 or 50 mg/kg, oral for 7 days). Data are presented as median ± IQR range; n = 4. Values of * and $ were significantly different from those of the control and CYC groups respectively, as found with the Kruskal–Wallis test followed by a Dunn multiple comparison test (p ˂ 0.05).

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  Figure 5. The impact of tenofovir (TFV) on renal and cardiac malondialdehyde (MDA) levels and total antioxidant capacity (TAC) altered via cyclophosphamide (CYC) injection. Data are expressed as mean ± standard error of the mean (SEM); n = 6. CYC (200 mg/kg, single i.p dose); TFV (50 mg/kg, oral for 7 days). The values of * and $ are significantly different from those of the control and CYC groups, respectively. This was determined using one-way ANOVA followed by Tukey–Kramer multiple comparison post hoc tests (p < 0.05).

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  Figure 6. The impact of tenofovir (TFV) on renal and cardiac AMP-activated protein kinase (AMPK), nuclear factor erythroid 2-related factor 2 (Nrf2), Heme oxygenase-1 (HO-1), B-cell lymphoma-2 (Bcl-2), and silent information regulator Sirtuin 1 (SIRT1) altered via cyclophosphamide (CYC) injection. (A): Protein levels in renal tissues. (B): Protein levels in cardiac tissues. Data are expressed as mean ± standard error of the mean (SEM); n = 6. CYC (200 mg/kg, single i.p dose); TFV (50 mg/kg, oral for 7 days). The values of * and $ are significantly different from those of the control and CYC groups, respectively. This was determined using one-way ANOVA followed by Tukey–Kramer multiple comparison post hoc tests (p < 0.05).

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  Figure 7. The impact of tenofovir (TFV) on renal and cardiac expression level of Interleukin-1B (IL-1B) and microtubule-associated protein 1A/1B-light chain 3 (LC3) altered via cyclophosphamide (CYC) injection. (A): IL-1B expression level in renal tissues. (B): IL-1B expression level in cardiac tissues. (A,a) Control group, (B,b) CYC group, (C,c) CYC + TFV 50 group, (D) column bar graph for the percentage of positive-stained area. Upper raw original magnification 10×, lower raw 40×, and scale bar 100 µm and 20 µm, respectively. Data are expressed as mean ± standard error of the mean (SEM); n = 4. CYC (200 mg/kg, single i.p dose); TFV (50 mg/kg, oral for 7 days). The values of * and $ are significantly different from those of the control and CYC groups, respectively. This was determined using one-way ANOVA followed by Tukey–Kramer multiple comparison post hoc tests (p < 0.05).

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  Figure 8. The impact of tenofovir (TFV) on renal and cardiac expression level of microtubule-associated protein 1A/1B-light chain 3 (LC3) altered via cyclophosphamide (CYC) injection. (A): LC3 expression level in renal tissues. (B): LC3 expression level in cardiac tissues. (A,a) Control group, (B,b) CYC group, (C,c) CYC + TFV 50 group, (D) column bar graph for percentage of positively stained area. Upper raw original magnification 10×, lower raw 40×, and scale bar 100 µm and 20 µm, respectively. Data are expressed as mean ± standard error of the mean (SEM); n = 4. CYC (200 mg/kg, single i.p dose); TFV (50 mg/kg, oral for 7 days). The values of * and $ are significantly different from those of the control and CYC groups, respectively. This was determined using one-way ANOVA followed by Tukey–Kramer multiple comparison post hoc tests (p < 0.05).