Xinkeshu Improves Endothelial Function and Augments Reendothelialization Capacity in Coronary Artery Disease with Anxiety/Depression

Figures

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  Figure 1. Flow chart. Patients received routine treatment combined with XKS or routine treatment for 1 month to assess the arterial function, anxiety/depressive symptoms, and the mechanisms.

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  Figure 2. The endothelial function and endothelial repair capacity of EPCs in CAD patients with or with anxiety/depression and healthy subjects. (a) The mean value of FMD (∗P < 0.01vs. healthy subjects, n = 30). (b) The mean value of migrated cell numbers (∗P < 0.01vs. the XKS group, n = 6), (c) the mean value of adhering cell numbers (∗P < 0.01vs. the XKS group, n = 6) of EPCs, and (d) the representative photographs of migration and adhesion function. (e) Representative photographs and (f) quantification analyses of reendothelialization areas (white) of the injured carotid arteries in nude mice 3 days after carotid denudation surgery plus intravenous injection of EPCs (5 × 105 cells) from CAD patients with or without anxiety/depression or healthy subjects (∗P < 0.01vs. healthy subjects, n = 6).

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  Figure 3. The effects of XKS on FMD, GAD-7, and PHQ-9. (a) The mean value of FMD (∗P < 0.05vs. baseline). (b) The mean value of GAD-7 score (∗∗P < 0.01vs. baseline). (c) The mean value of PHQ-9 score (∗∗P < 0.01vs. baseline).

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  Figure 4. XKS improves the in vitro functions and in vivo endothelial repair capacity of EPCs. (a) The representative photographs of migration function and (b) the mean value of migrated cell numbers of EPCs incubated among different concentrations of XKS (∗P < 0.05 and ∗∗P < 0.01vs. the vehicle group, n = 6). (c) The representative photographs of adhesion function and (d) the mean value of adhering cell numbers of EPCs incubated among different concentrations of XKS (∗P < 0.05 and ∗∗P < 0.01vs. the vehicle group, n = 6). (e) Representative photographs and quantification analyses of reendothelialization areas (white) of the injured carotid arteries in nude mice 3 days after carotid denudation surgery plus intravenous injection of EPCs (5 × 105 cells) with or without XKS (∗∗P < 0.01vs. the non-XKS group, n = 6). (f) The pathological changes of endothelium from carotid arteries in hematoxylin-eosin (HE) and immunohistochemistry (IHC) staining of CD31 (magnification: 400).

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  Figure 5. XKS regulates CXCR7/p38/cleaved casepase-3 pathway and inhibits apoptosis of EPCs from CAD patients with anxiety/depression. (a) The quantification analyses of CXCR7 mRNA (∗P < 0.05vs. the vehicle group, n = 6). (b) The protein bands of CXCR7 and GAPDH and the mean value of CXCR7/GAPDH ratio (∗P < 0.05 and ∗∗P < 0.01vs. the vehicle group, n = 6). (c) The protein bands of CXCR7 and GAPDH after transfecting with siRNA-CXCR7 and the mean value of CXCR7/GAPDH ratio (∗P < 0.01vs. the non-XKS group, n = 6). (d) The protein bands of p-p38, p38, and GAPDH and the mean value of p-p38/p38 ratio (∗P < 0.01vs. the blank group, n = 6). (e) The protein bands of cleaved casepase-3 and GAPDH and the mean value of cleaved casepase-3/GAPDH ratio (∗P < 0.01vs. the non-XKS group, n = 6). (f) The quantification analyses of apoptosis cells rate measured by flow cytometry (∗P < 0.01vs. the XKS group; n = 3 per group).

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  Figure 6. XKS-mediated CXCR7 signaling improves in vitro functions and in vivo endothelial repair capacity of EPCs. (a) The representative photographs of migration and adhesion function, (b) the mean value of migrated cell numbers (∗P < 0.01vs. the non-XKS group, n = 6), and (c) the mean value of adhering cell numbers (∗P < 0.01vs. the non-XKS group, n = 6) of EPCs. (d) The quantification analyses and (e) representative photographs of reendothelialization areas (white) of the injured carotid arteries in nude mice 3 days after carotid denudation surgery plus intravenous injection of EPCs (5 × 105 cells) with or without XKS (∗P < 0.01vs. the non-XKS group, n = 6). (f) The pathological changes of endothelium from carotid arteries in hematoxylin-eosin (HE) and immunohistochemistry (IHC) staining of CD31 (magnification: 400).