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Overexpression of SerpinE2/protease nexin-1 Contribute to Pathological Cardiac Fibrosis via increasing Collagen Deposition

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ABSTRACT

Although increases in cardiovascular load (pressure overload) are known to elicit ventricular remodeling including cardiomyocyte hypertrophy and interstitial fibrosis, the molecular mechanisms of pressure overload or AngII -induced cardiac interstitial fibrosis remain elusive. In this study, serpinE2/protease nexin-1 was over-expressed in a cardiac fibrosis model induced by pressure-overloaded via transverse aortic constriction (TAC) in mouse. Knockdown of serpinE2 attenuates cardiac fibrosis in a mouse model of TAC. At meantime, the results showed that serpinE2 significantly were increased with collagen accumulations induced by AngII or TGF-β stimulation in vitro. Intriguingly, extracellular collagen in myocardial fibroblast was reduced by knockdown of serpinE2 compared with the control in vitro. In stark contrast, the addition of exogenous PN-1 up-regulated the content of collagen in myocardial fibroblast. The MEK1/2- ERK1/2 signaling probably promoted the expression of serpinE2 via transcription factors Elk1 in myocardial fibroblast. In conclusion, stress-induced the ERK1/2 signaling pathway activation up-regulated serpinE2 expression, consequently led accumulation of collagen protein, and contributed to cardiac fibrosis.

No MeSH data available.


The changes of collagen content induced by exogenous serpinE2.(a) Differential expression of serpinE2 in myocardial fibroblast and myocardial cell. n = 3. *P < 0.05 vs. control. Full-length blots/gels are presented in Supplementary Figure 6. (b) Relative level of collagen in supernatants of fibroblast by over-expression of serpinE2 or knock-down of serpinE2. n = 6. *P < 0.05 vs. control, #P < 0.05 vs. serpinE2 group. (c) The Immunofluorescence for serpinE2 in myocardial fibroblast. Scale bar:10 μm.
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f4: The changes of collagen content induced by exogenous serpinE2.(a) Differential expression of serpinE2 in myocardial fibroblast and myocardial cell. n = 3. *P < 0.05 vs. control. Full-length blots/gels are presented in Supplementary Figure 6. (b) Relative level of collagen in supernatants of fibroblast by over-expression of serpinE2 or knock-down of serpinE2. n = 6. *P < 0.05 vs. control, #P < 0.05 vs. serpinE2 group. (c) The Immunofluorescence for serpinE2 in myocardial fibroblast. Scale bar:10 μm.

Mentions: The histological structure of the heart is mainly composed of mast myocardial cells and myocardial fibroblasts. SerpinE2 is both expressed in myocardial fibroblast and myocardial cell as well, moreover, the expression of serpinE2 in the myocardial fibroblast was higher than that in myocardial cell (Fig. 4a). Subcellular locations from UniProtKB/Swiss-Prot database for SERPINE2 Gene showed that serpinE2 is mostly secreted into extracellular space, next is in cytosol. Since serpinE2 can be secreted by fibroblasts into extracellular fluid14, the exogenous serpinE2 (serpinE2-gst Fusion Protein) was added to cardiac fibroblasts at final concentration of 10 ng/ml for 24 h and 48 h. Then the level of collagen were all increased significantly about 1.33-fold and 1.95-fold compared with the control in supernatant of fibroblasts detected by Sircol™ Collagen Assay at 24 h and 48 h (Fig. 4b). On the contrary, when cardiac fibroblasts was knock-down of serpinE2 with serpinE2 shRNA for 24 h and 48 h, the content of collagen was obviously reduced compared with treated with serpinE2 group (Fig. 4b). The immunofluorescence showed the expression of serpinE2, when the exogenous serpinE2 was added or knock-down of serpinE2 in myocardial fibroblast. The expression of serpinE2 is increased in fibroblasts treated with exogenous serpinE2 than that in the control (Fig. 4c). It illustrated the exogenous serpinE2 can entry into the fibroblasts by some way. In addition it was supported by the hypothesis that serpinE2/ protease nexin-1 can induce endocytosis binding to uPA/uPAR complexes mediated by LRP2223.


Overexpression of SerpinE2/protease nexin-1 Contribute to Pathological Cardiac Fibrosis via increasing Collagen Deposition
The changes of collagen content induced by exogenous serpinE2.(a) Differential expression of serpinE2 in myocardial fibroblast and myocardial cell. n = 3. *P < 0.05 vs. control. Full-length blots/gels are presented in Supplementary Figure 6. (b) Relative level of collagen in supernatants of fibroblast by over-expression of serpinE2 or knock-down of serpinE2. n = 6. *P < 0.05 vs. control, #P < 0.05 vs. serpinE2 group. (c) The Immunofluorescence for serpinE2 in myocardial fibroblast. Scale bar:10 μm.
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f4: The changes of collagen content induced by exogenous serpinE2.(a) Differential expression of serpinE2 in myocardial fibroblast and myocardial cell. n = 3. *P < 0.05 vs. control. Full-length blots/gels are presented in Supplementary Figure 6. (b) Relative level of collagen in supernatants of fibroblast by over-expression of serpinE2 or knock-down of serpinE2. n = 6. *P < 0.05 vs. control, #P < 0.05 vs. serpinE2 group. (c) The Immunofluorescence for serpinE2 in myocardial fibroblast. Scale bar:10 μm.
Mentions: The histological structure of the heart is mainly composed of mast myocardial cells and myocardial fibroblasts. SerpinE2 is both expressed in myocardial fibroblast and myocardial cell as well, moreover, the expression of serpinE2 in the myocardial fibroblast was higher than that in myocardial cell (Fig. 4a). Subcellular locations from UniProtKB/Swiss-Prot database for SERPINE2 Gene showed that serpinE2 is mostly secreted into extracellular space, next is in cytosol. Since serpinE2 can be secreted by fibroblasts into extracellular fluid14, the exogenous serpinE2 (serpinE2-gst Fusion Protein) was added to cardiac fibroblasts at final concentration of 10 ng/ml for 24 h and 48 h. Then the level of collagen were all increased significantly about 1.33-fold and 1.95-fold compared with the control in supernatant of fibroblasts detected by Sircol™ Collagen Assay at 24 h and 48 h (Fig. 4b). On the contrary, when cardiac fibroblasts was knock-down of serpinE2 with serpinE2 shRNA for 24 h and 48 h, the content of collagen was obviously reduced compared with treated with serpinE2 group (Fig. 4b). The immunofluorescence showed the expression of serpinE2, when the exogenous serpinE2 was added or knock-down of serpinE2 in myocardial fibroblast. The expression of serpinE2 is increased in fibroblasts treated with exogenous serpinE2 than that in the control (Fig. 4c). It illustrated the exogenous serpinE2 can entry into the fibroblasts by some way. In addition it was supported by the hypothesis that serpinE2/ protease nexin-1 can induce endocytosis binding to uPA/uPAR complexes mediated by LRP2223.

View Article: PubMed Central - PubMed

ABSTRACT

Although increases in cardiovascular load (pressure overload) are known to elicit ventricular remodeling including cardiomyocyte hypertrophy and interstitial fibrosis, the molecular mechanisms of pressure overload or AngII -induced cardiac interstitial fibrosis remain elusive. In this study, serpinE2/protease nexin-1 was over-expressed in a cardiac fibrosis model induced by pressure-overloaded via transverse aortic constriction (TAC) in mouse. Knockdown of serpinE2 attenuates cardiac fibrosis in a mouse model of TAC. At meantime, the results showed that serpinE2 significantly were increased with collagen accumulations induced by AngII or TGF-&beta; stimulation in vitro. Intriguingly, extracellular collagen in myocardial fibroblast was reduced by knockdown of serpinE2 compared with the control in vitro. In stark contrast, the addition of exogenous PN-1 up-regulated the content of collagen in myocardial fibroblast. The MEK1/2- ERK1/2 signaling probably promoted the expression of serpinE2 via transcription factors Elk1 in myocardial fibroblast. In conclusion, stress-induced the ERK1/2 signaling pathway activation up-regulated serpinE2 expression, consequently led accumulation of collagen protein, and contributed to cardiac fibrosis.

No MeSH data available.