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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.

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ANG-II-mediated increase in cardiac fibrosis and the serpinE2 expression.(a) qRT-PCR assay was applied for the detection of collagen I and collagen III mRNA expression in myocardial fibroblasts (n = 6, *P < 0.05 vs. control) in 50 nM AngII induced collagen deposition; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblast induced by AngII (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (d) Western blot analysis of serpinE2 protein (left panel) and statistical analysis (right panel) (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 4; (e) qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by AngII (n = 8, *P < 0.01 vs. control).
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f2: ANG-II-mediated increase in cardiac fibrosis and the serpinE2 expression.(a) qRT-PCR assay was applied for the detection of collagen I and collagen III mRNA expression in myocardial fibroblasts (n = 6, *P < 0.05 vs. control) in 50 nM AngII induced collagen deposition; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblast induced by AngII (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (d) Western blot analysis of serpinE2 protein (left panel) and statistical analysis (right panel) (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 4; (e) qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by AngII (n = 8, *P < 0.01 vs. control).

Mentions: AngII stimulation is linked to cardiac remodeling characterized by fibrosis with collagen accumulation. Consistently, as the result, the mRNA level of Collagen I and III were obviously increased in myocardial fibroblast after 24 h and 48 h treatment with 50 nM AngII and compared with the control group (Fig. 2a). Total contents of collagen were increased in fibroblast and in the supernatants of fibroblast (Fig. 2b), suggesting that ANG-II-mediated the secretion of collagen from fibroblasts into ECM. According to database of UniProtKB and Swiss-Prot for SERPINE2 Gene, serpinE2 is a kind of protein that is more likely to be secreted into the extracellular space, rather than or less into cytosol. Therefore, we have a reason to believe that the serpinE2 may play a certain function at the cell membrane and it can could be detected in the culture medium1011. Based upon this regard, we then detected serpinE2 content in the supernatants of myocardial fibroblasts by ELISA assay. Compared with the control group, serpinE2 level was significantly increased about 1.63-fold and 1.92-fold, respectively, after 24 h and 48 h treatment with 50 nM AngII (Fig. 2c). It means that AngII very likely to promote serpinE2 secretion from fibroblasts into the supernatants. The data from western blot and qRT-PCR also showed a corresponding increment of serpinE2 in myocardial fibroblasts (Fig. 2d and e).


Overexpression of SerpinE2/protease nexin-1 Contribute to Pathological Cardiac Fibrosis via increasing Collagen Deposition
ANG-II-mediated increase in cardiac fibrosis and the serpinE2 expression.(a) qRT-PCR assay was applied for the detection of collagen I and collagen III mRNA expression in myocardial fibroblasts (n = 6, *P < 0.05 vs. control) in 50 nM AngII induced collagen deposition; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblast induced by AngII (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (d) Western blot analysis of serpinE2 protein (left panel) and statistical analysis (right panel) (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 4; (e) qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by AngII (n = 8, *P < 0.01 vs. control).
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5120308&req=5

f2: ANG-II-mediated increase in cardiac fibrosis and the serpinE2 expression.(a) qRT-PCR assay was applied for the detection of collagen I and collagen III mRNA expression in myocardial fibroblasts (n = 6, *P < 0.05 vs. control) in 50 nM AngII induced collagen deposition; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblast induced by AngII (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (d) Western blot analysis of serpinE2 protein (left panel) and statistical analysis (right panel) (n = 6, *P < 0.01 vs. control). Full-length blots/gels are presented in Supplementary Figure 4; (e) qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by AngII (n = 8, *P < 0.01 vs. control).
Mentions: AngII stimulation is linked to cardiac remodeling characterized by fibrosis with collagen accumulation. Consistently, as the result, the mRNA level of Collagen I and III were obviously increased in myocardial fibroblast after 24 h and 48 h treatment with 50 nM AngII and compared with the control group (Fig. 2a). Total contents of collagen were increased in fibroblast and in the supernatants of fibroblast (Fig. 2b), suggesting that ANG-II-mediated the secretion of collagen from fibroblasts into ECM. According to database of UniProtKB and Swiss-Prot for SERPINE2 Gene, serpinE2 is a kind of protein that is more likely to be secreted into the extracellular space, rather than or less into cytosol. Therefore, we have a reason to believe that the serpinE2 may play a certain function at the cell membrane and it can could be detected in the culture medium1011. Based upon this regard, we then detected serpinE2 content in the supernatants of myocardial fibroblasts by ELISA assay. Compared with the control group, serpinE2 level was significantly increased about 1.63-fold and 1.92-fold, respectively, after 24 h and 48 h treatment with 50 nM AngII (Fig. 2c). It means that AngII very likely to promote serpinE2 secretion from fibroblasts into the supernatants. The data from western blot and qRT-PCR also showed a corresponding increment of serpinE2 in myocardial fibroblasts (Fig. 2d and e).

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.


Related in: MedlinePlus