The Cardioprotective Potential of Red Onion Waste Against Isoprenaline‐Induced Myocardial Infarction in Rat: Molecular Docking and In Vivo Studies

Figures

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  FIGURE 1. Experimental design. ISO: isoprenaline, MI: Myocardial inarction, NS: normal saline.

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  FIGURE 2. DPPH, FRAP, and ABTS in vitro antioxidant activities of red onion scales.

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  FIGURE 3. Effect of red onion scales extract on echocardiographic alterations in isoprenaline‐induced MI in rats. Figures (A‐E) represent M‐mode imaging of (A) Control, (B) Red onion scales 300, (C) ISO, (D) Red onion 150+ISO, (E) Red onion 300+ISO, and (F) EF% Red onion scales extract was introduced orally at doses of 150 and 300 mg/kg for 30 days, while ISO (65 mg/kg; s.c.) was injected on days 29 and 30 to induce MI. Results were illustrated as mean ± standard deviation (SD) (n = 8 per group). Data was analyzed using one‐way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test. The significant differences were set at (p < 0.05) among all groups. a significantly different from the control group, b significantly different from the ISO group. EF: Ejection fraction; ISO: Isoprenaline; MI: Myocardial infarction.

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  FIGURE 4. Effect of red onion scales extract on serum MI markers; (A) CK‐MB and (B) Troponin T in isoprenaline‐induced MI in rats.

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  FIGURE 5. Effect of red onion scales extract on inflammatory markers; Figures (A‐F) represent NF‐κB (p65) expression. The photomicrographs exhibited the expression as the following: (A) Control, (B) Red onion 300, (C) ISO, (D) Red onion 150+ISO, (E) Red onion 300+ISO, and (F) The immunohistochemical expression as area%, as well as (G) IL‐1β. For 30 days, red onion scales extract was administered (150/300 mg/kg; p.o.), while ISO (65 mg/kg; s.c.) was injected on days 29 and 30 to induce MI, Results were presented as mean ± standard deviation (n = 4 per group). Data was analyzed using one‐way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test. The significant differences were set at (p < 0.05) among all groups. a significantly different from the control group, b significantly different from the ISO group, and c significantly different from the red onion 150+ISO group. IL‐1β: Interleukin‐1 beta; ISO: Isoprenaline; MI: Myocardial infarction; NF‐κB: Nuclear factor kappa B.

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  FIGURE 6. Effect of red onion scales extract on iNOS expression in isoprenaline‐induced MI in rats. Figures (A‐F) represent iNOS expression; (A) Control, (B) Red onion 300, (C) ISO, (D) Red onion 150+ISO, (E) Red onion 300+ISO, and (F) The immunohistochemical expression represented as area%. Red onion scales extract was administered (150/300 mg/kg; p.o.) for 30 days, while ISO (65 mg/kg; s.c.) was injected on days 29 and 30 to induce MI. Results were displayed as mean ± standard deviation (n = 3 per group). Data were analyzed using one‐way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test. The significant differences were set at (p < 0.05) among all groups. a significantly different from the control group, b significantly different from the ISO group. iNOS: inducible nitric oxide synthases; ISO: Isoprenaline; MI: Myocardial infarction.

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  FIGURE 7. Effect of red onion scales extract on antioxidant/oxidant markers; (A) Nrf2, (B) SOD, and (C) MDA in isoprenaline‐induced MI in rats. Red onion scales extract was administered (150/300 mg/kg; p.o.) for 30 days, while twice injections of ISO (65 mg/kg; s.c.) were carried out on day 29 and 30 to induce MI insult. Results were represented as mean ± standard deviation (SD) (n = 4 per group). Data were analyzed using one‐way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test. The significant differences were set at (p < 0.05) among all groups. a significantly different from the control group, bsignificantly different from the ISO group. ISO: Isoprenaline; MDA: Malondialdehyde; MI: Myocardial infarction; Nrf2: Nuclear factor erythroid‐ 2; SOD: Superoxide dismutase.

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  FIGURE 8. Effect of red onion scales extract on apoptotic marker in isoprenaline‐induced MI in rats; Figures (A‐F) represent caspase 3 expression;(A) Control, (B) Red onion 300, (C) ISO, (D) Red onion 150+ISO, (E) Red onion 300+ISO, and (F) The immunohistochemical expression shown as area%. Red onion scales extract was received orally (150/300 mg/kg) for 30 days, while ISO (65 mg/kg; s.c.) was injected on day 29 and 30 to provoke MI. Results were displayed as mean ± standard deviation (SD) (n = 3 per group). Data was analyzed using one‐way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test. The significant differences were set at (p < 0.05) among all groups. a significantly different from the control group, b significantly different from the ISO group, and c significantly different from the red onion 150+ISO group. ISO: Isoprenaline; MI: Myocardial infarction.

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  FIGURE 9. Effect of red onion scales extract on endoplasmic reticulum stress markers; (A) ATF4 and (B) CHOP in isoprenaline‐induced MI in rats. Red onion scales extract was administered (150/300 mg/kg; p.o.) for 30 days, while ISO (65 mg/kg; s.c.) was injected on day 29 and 30 to induce MI. Results were displayed as mean ± standard deviation (n = 4 per group). Data were analyzed using one‐way analysis of variance (ANOVA) followed by Tukey's multiple comparisons test. The significant differences were set at (p < 0.05) among all groups. a significantly different from the control group, b significantly different from the ISO group, and c significantly different from the red onion 150+ISO group. ATF4: Activating transcription factor 4; CHOP: C/EBP homologous protein; ISO: Isoprenaline; MI: Myocardial infarction.

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  FIGURE 10. Effect of red onion scales extract on histopathological changes in isoprenaline‐induced MI in rats: Photomicrograph of cardiac tissue sections showed control group (A) exhibited the standard normal architecture of cardiomyocytes. They seem branched, elongated, and cross‐striated with large oval central nuclei (thick arrow) associated with narrow slit‐like interstices (arrowhead). Moreover, Red onion 300 group (B) demonstrated structure nearly resembles the negative control group that evidenced by the intact cardiomyocytes with large oval central nuclei (thick arrow) and narrow slit‐like interstices (arrowhead) in between. In contrast, ISO group (C & D) highlighted intense histopathological alterations including degeneration and hyalinization of muscle fibers (rectangle), interstitial hemorrhage at the interstices between cardiac myofibers (arrowhead), severe vascular congestion (star), marked inflammatory cell infiltration (wave arrow), apoptotic assembly of cardiomyocytes (arrows), distinct edema leading to dispersion between muscle fiber (curve arrow), noticeable replacing of muscle fiber with connective tissue fibers (circle) along with obvious expanse in fibers quantity by…

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  FIGURE 11. 3D‐representation (left) and 2D‐ligand interactions (right) of quercetin in the active site of MPO (PDB: 4C1M).

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  FIGURE 12. Schematic presentation of the cardioprotective effects of red onion scales extract. ATF4: Activating transcription factor 4. CHOP: C/EBP homologous protein. IL‐1β: Interleukin‐1 beta. ISO: Isoprenaline. NF‐κB: Nuclear factor‐kappa B.