Endothelial‑to‑mesenchymal transition in human idiopathic dilated cardiomyopathy - PubMed (original) (raw)

Endothelial‑to‑mesenchymal transition in human idiopathic dilated cardiomyopathy

Yeqing Xie et al. Mol Med Rep. 2018 Jan.

Abstract

Dilated cardiomyopathy (DCM) is characterized by left ventricular dilation and cardiac fibrosis. Emerging evidence indicated that endothelial‑to‑mesenchymal transition (Endo‑MT) is a crucial event during organ fibrosis. This study was performed to clarify whether Endo‑MT contributed to the progression of cardiac fibrosis in DCM. Cardiac samples from patients with DCM and control were obtained. The presence of endothelial markers, cluster of differentiation (CD)31 and vascular endothelial (VE)‑cadherin, and mesenchymal markers, α smooth muscle actin (SMA) and fibroblast‑specific protein 1 (FSP1) was performed using immunohistochemistry. Co‑localization of endothelial markers and mesenchymal markers were identified using confocal immunofluorescence staining. Serum procollagen type I carboxy‑terminal propeptide (PICP) and procollagen type III amino‑terminal propeptide (PIIINP) were measured by ELISA. Protein levels of Wnt, β‑catenin and Snail were determined using western blot analysis. Immunohistochemistry and double‑immunofluorescence staining demonstrated that the expression of CD31 and VE‑cadherin were significantly decreased in DCM samples, whereas the FSP‑1, and αSMA were significantly increased. CD31 and VE‑cadherin labeling indexes were respectively negatively correlated with left ventricular end‑diastolic diameter (LVEDD) (CD31 r=‑0.82, P<0.01; VE‑cadherin r=-0.73, P<0.01), while FSP‑1 and αSMA were positively associated with LVEDD (αSMA r=0.65, P<0.01, FSP1 r=0.53, P<0.01) and left ventricular ejection fraction (αSMA r=‑0.18, P<0.05; FSP1 r=‑0.21, P<0.05). Furthermore, PICP and PIIINP levels were positively associated with the co‑expression labeling indexes (CD31/SMA co‑labeling index and PICP r=0.727, P<0.01; CD31/SMA co‑labeling index and PIIINP r=0.741, P<0.01; VE‑Cadherin/FSP‑1 co‑labeling index and PICP r=0.716, P<0.01; VE‑cadherin/FSP‑1 co‑labeling index and PIIINP r=0.648, P<0.05). Western blot analysis indicated that proteins levels of Wnt signaling and snail were significantly increased in DCM samples. These results suggested that Endo‑MT is potentially implicated in the pathogenesis of myocardial fibrosis and remodeling during the development of DCM, indicating a potential therapeutic target for DCM treatment.

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Figures

Figure 1.

Figure 1.

Cardiac fibrosis and remodeling in recruit patients: (A) Pathological features of enrolled sample: Increased amount of interstitial fibrosis was observed in DCM samples comparing control as indicated by HE and Sirius Red staining. (B) Serum PICP and (C) PIIIINP levels were significantly increased in DCM patient comparing control (P<0.05).

Figure 2.

Figure 2.

(A) Representative image and (B) immunostaining of endothelial and mesenchymal markers in both normal and pathologic cardiac sections: Compared with control, the expression of αSMA and FSP1 was increased in DCM specimen, whereas expression of CD31 and VE-cadherin was reduced (P<0.05).

Figure 3.

Figure 3.

Co-localization of endothelial and mesenchumal markers in human cardiac samples: Double inmmunofluorescence staining of CD31 and αSMA, and VE-cadherin and FSP1 was performed for normal heart and DCM cardiac samples. The expression of CD31 and VE-cadherin were colored with green, and the expression of αSMA and FSP1 were colored with red. Nuclei were stained with 4′6-diamidino-2-phenylindole (DAPI). Coexpression of CD31 and αSMA was observed in cardiac endothelial cells of DCM, as well as VE-cadherin and FSP1.

Figure 4.

Figure 4.

Relationship between cardiac functions and immunostaining indexes of endothelial and mesenchymal markers in DCM patients: Cardiac functions were demonstrated as left ventricular end diastolic diameters (LVEDD) and left ventricular ejection fraction (EF). CD31 and VE-cadherin immunestaining labeling indexes were respectively negatively correlated with left ventricular end-diastolic diameter (A) CD31 r=−0.82, P<0.01; VE-cadherin r=−0.73, P<0.01, while FSP-1 and αSMA immunestaining labeling indexes were positively associated with left ventricular chamber enlargement (B) αSMA r=0.65, P<0.01, FSP1 r=0.53, P<0.01 and left ventricular ejection fraction (D) αSMA r=−0.18, P<0.05; FSP1 r=−0.21, P<0.05. However, neither CD31 nor VE-cadherin has statistically significant correlation with left ventricular ejection fraction (C) CD31 r=0.16, VE-cadherin r=0.24, P>0.05.

Figure 5.

Figure 5.

Relationship between serum levels of PICP and PIIINP and co-localization labeling indexes in DCM patients: Both circulating PICP and PIIINP levels were positively related with the co-expression labeling indexes. (A) CD31/SMA co-labeling index and PICP r=0.727, P<0.01; (B) CD31/SMA co-labeling index and PIIINP r=0.741, P<0.01; (C) CD31/SMA co-labeling index and PICP r=0.727, P<0.01; VE-Cadherin/FSP-1 co-labeling index and PICP r=0.716, P<0.01; (D) VE-cadherin/FSP-1 co-labeling index and PIIINP r=0.648, P<0.05).

Figure 6.

Figure 6.

Wnt signaling pathway in DCM: (A) Western blot analysis for the expression of Wnt signaling proteins was performed on protein isolated from hearts of normal donors and DCM patients. (B) Quantification of proteins showed that conical Wnt-related protein was significantly increased in DCM specimen, compared with control (P<0.05).

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