Qishen capsule safely boosts cardiac function and angiogenesis via the MEK/ERK pathway in a rat myocardial infarction model.

Figures

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  Figure 1.. Echocardiography and hemodynamics assessment of cardiac function. (A): Representative images of echocardiography at the baseline and the endpoint; (B): echocardiography measurement of LVEF; (C): echocardiography measurement of LVFS at the endpoint; (D): hemodynamics assessment of dp/dtmax; and (E): hemodynamics assessment of LVEDP at the endpoint (n = 5 in each group). The baseline is four weeks after infarction, and the endpoint is eight weeks after infarction. *P < 0.05 compared with the Sham group, #P < 0.05 compared with the MI group. dp/dtmax: left ventricular pressure maximal rate of rise; LVEDP: left ventricular end-diastolic pressure; LVEF: left ventricular ejection fraction; LVFS: left ventricular fractional shortening; MI: myocardial infarction; QS: Qishen capsule.

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  Figure 2.. Quantitative histological analysis and viable myocardium identification. (A): Representative heart weight index; (B): quantitative analysis of infarct scar size; (C): quantitative analysis of infarct scar area; (D): quantitative analysis of thinning ratio with Masson's trichrome staining; and (E): representative immunofluorescence of viable myocardium positive for desmin and α-actinin in the infarct area. n = 6 in each group. The magnification is 400 ×. *P < 0.05 compared with the Sham group, #P < 0.05 compared with the MI group. MI: myocardial infarction; QS: Qishen capsule.

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  Figure 3.. Histopathological staining. (A): Inflammatory cell infiltration shown with Hematoxylin-eosin staining; (B): fibrosis shown with Masson's trichrome staining (12.5 ×); (C): fibrosis shown with Masson's trichrome staining (400 ×); (D): picrosirius red staining to detect the collagen content within the infarct zone (12.5 ×); and (E): picrosirius red staining to detect the collagen content within the infarct zone (400 ×). n = 3 in each group. MI: myocardial infarction; QS: Qishen capsule.

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  Figure 4.. Ultrastructural analysis of the myocardial and capillary architecture. (A): Representative transmission electron microscopy images of the mitochondrial morphology. Mitochondrial swelling with loss of cristae and mitochondria changed to vacuoles were detected in the MI group; and (B): representative transmission electron microscopy images of the capillary architecture, and we found endothelial edema and endoplasmic reticulum expansion in the MI group. MI: myocardial infarction; QS: Qishen capsule.

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  Figure 5.. Immunohistochemistry and immunofluorescence assessment of myocardial angiogenesis. (A): Immunofluorescence for the expression of angiogenesis positive for VEGF and α-SMA in the vascular endothelium; (B): sections were stained with α-SMA staining for the arteriolar density measurement; (C): quantification of the arteriolar density; (D): CD31 immunostaining to identify capillaries in the peri-infarct region; (E): CD31 immunostaining to identify capillaries in the infarct region; and (F): quantification of the capillary density. n = 6 in per group. The magnification for A is 400 ×, for B, D and E is 200 ×. *P < 0.05 compared with the Sham group, #P < 0.05 compared with the MI group. MI: myocardial infarction; QS: Qishen capsule.

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  Figure 6.. QS treatment increased the levels of VEGF and HIF-1α. (A): The protein level of VEGF from enzyme-linked immunosorbent assay; and (B): the mRNA levels of VEGF and HIF-1α from quantitative real-time polymerase chain reaction. *P < 0.05 compared with the Sham group, #P < 0.05 compared with the MI group. HIF-1α: hypoxia inducible factor-1α; MI: myocardial infarction; QS: Qishen capsule; VEGF: vascular endothelial growth factor.

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  Figure 7.. The activities of the Akt and MEK/ERK1/2 pathways were detected with western blotting. (A): The phosphorylation of MEK and ERK1/2 was attenuated markedly by QS in the post-infarct hearts, whereas the phosphorylation of Akt increased; and (B): the histogram shows the relative protein levels of the phosphorylation of Akt and MEK/ERK1/2. The data represent the results of three separate experiments. The results are expressed as the means ± SD. *P < 0.05 compared with the Sham group, #P < 0.05 compared with the MI group. Akt: protein kinase B; ERK: extracellular regulated protein kinase; GAPDH: glyceraldehyde-3-phosphate dehydrogenase; MEK: mitogen-activated protein kinase; MI: myocardial infarction; QS: Qishen capsule.