Integrating network pharmacology and experimental validation to investigate the mechanism of Gualou Xiebai Banxia decoction against myocardial ischemia

Figures

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  Figure 1. Overlapped targets screen. (A) Overlapping targets of overlapping GXBD and MI. Blue circle represented GXBD targets and yellow circle represented MI targets. The overlapping region was the target of GXBD against MI. (B) The protein-protein interaction (PPI) network of targets. PPI network of potential targets of GXBD against MI. (C) Relationship network between potential therapeutic targets against MI and active components of GXBD (The light red triangle represented the active components of Gualou, the green triangle represented the active components of Xiebai, the blue triangle represented the active components of Banxia, the dark red triangle represented the common components of Banxia and Xiebai, and the blue square represented the target of Gualou Xiebai Banxia Decoction for anti-myocardial ischemia).

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  Figure 2. Results of GO enrichment and KEGG pathway analyses of GXBD targets. (A) Biological process. (B) Cellular component. (C) Molecular function. (D) KEGG pathway. (The GO terms are located on the Y-axis, and the numbers on the X-axis indicate the degree of enrichment. Red indicated high reliability, while blue indicated low reliability.).

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  Figure 3. GXBD ameliorated cardiac function in MI rats. (A) Electrocardiogram of rats. (B) Representative M-mode echocardiography images. (C–F) EF ejection fraction, FS fractional shortening of the left ventricular diameter, LVPED diastolic left ventricular internal diameter, LVPWS systolic left ventricular posterior wall thickness. (G) Heart-to-Body-Weight ratio. All data were expressed as mean ± SD, n = 5. Compared with the control group, #P < 0.05, ##P < 0.01. Compared with the MI model group, *P < 0.05, **P < 0.01.

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  Figure 4. GXBD improved cardiac injury in MI rats. (A) Cardiac ischemia was detected by TTC staining. (B) H&E staining was used to observe the pathological changes of rat heart tissue (×400). (C) Infarction area. (D) brain natriuretic peptide. (E) creatine kinase isoenzyme. (F) lactate dehydrogenase. All data were expressed as mean ± SD, n = 5. Compared with the control group, #P < 0.05, ##P < 0.01. Compared with the MI model group, *P < 0.05, **P < 0.01.

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  Figure 5. GXBD reduced oxidative stress in MI rats. (A) superoxide dismutase. (B) malondialdehyde. (C) glutathione peroxidase. (D) ROS content in rat myocardial tissue (×200). All data were expressed as mean ± SD, n = 5. Compared with the control group, #P < 0.05, ##P < 0.01. Compared with the MI model group, *P < 0.05, **P < 0.01.

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  Figure 6. GXBD improved mitochondrial function in MI rats. (A) Typical cardiac images drawn in cardiac sections using JC-1 staining (×200). (B) ATP. (C) MFN2 and DRP1 protein expression in cardiac tissues of rats. (D) MFN2/β-actin ratio. (E) DRP1/β-actin ratio. All data were expressed as mean ± SD, n = 5. Compared with the control group, #P < 0.05, ##P < 0.01. Compared with the MI model group, *P < 0.05, **P < 0.01.

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  Figure 7. Effect of GXBD on PI3K/AKT/NRF2 pathway protein expression in cardiac tissue of rats with MI. (A) p-PI3K, PI3K, p-AKT and AKT protein expression in cardiac tissues of rats. (B) p-PI3K/PI3K ratio. (C) p-AKT/AKT ratio. (D) Nuclear NRF2, Cytoplasmic NRF2, NRF2 and HO-1 protein expression in cardiac tissues of rats. (E) NRF2/Lamin B ratio. (F) Cytoplasmic NRF2/β-actin ratio. (G) NRF2/β-actin ratio. (H) HO-1/β-actin ratio. All data were expressed as mean ± SD, n = 3. Compared with the control group, #P < 0.05,##P < 0.01. Compared with the MI model group, *P < 0.05, **P < 0.01.

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  Figure 8. Effects of GXBD on nuclear transcription of NRF2 in MI rats (×400) (n = 5).