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

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-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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TGF-β-induced fibrosis and mediated increase in serpinE2 expression.(a) qRT-PCR assay was applied for detection of collagen I and collagen III mRNA expression in myocardial fibroblast (n = 6, *P < 0.05 vs. control) in TGF-β-induced fibrosis; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblast (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 5; (e). qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by TGF-β (n = 8, *P < 0.01 vs. control).
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f3: TGF-β-induced fibrosis and mediated increase in serpinE2 expression.(a) qRT-PCR assay was applied for detection of collagen I and collagen III mRNA expression in myocardial fibroblast (n = 6, *P < 0.05 vs. control) in TGF-β-induced fibrosis; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblast (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 5; (e). qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by TGF-β (n = 8, *P < 0.01 vs. control).

Mentions: It has well been accepted that the activation of renin-angiotensin-aldosterone system would increase levels of active TGF-β that plays an important function in pressure overload-induced cardiac fibrosis20 and lead to increase in the mRNA expressions of collagen I and III in myocardial fibroblasts2021. Compared with the control group, the total collagen concentration detected by Sircol™ Collagen Assay were significantly elevated in myocardial fibroblasts after 24 h and 48 h stimulation with TGF-β 10 ng/ml (Fig. 3a and b), meanwhile, quantified serpinE2 secretion by ELISA assay into the supernatants of fibroblast culture medium showed accordant changes with collagen content. These results indicate that TGF-β also promotes serpinE2 secretion from fibroblasts into the supernatants (Fig. 3c). The supportive results were accordingly also obtained using western blot and qRT-PCR showing the up-regulation of both protein and mRNA expression of serpinE2 in myocardial fibroblasts treated with TGF-β (Fig. 3d and e).


Overexpression of SerpinE2/protease nexin-1 Contribute to Pathological Cardiac Fibrosis via increasing Collagen Deposition
TGF-β-induced fibrosis and mediated increase in serpinE2 expression.(a) qRT-PCR assay was applied for detection of collagen I and collagen III mRNA expression in myocardial fibroblast (n = 6, *P < 0.05 vs. control) in TGF-β-induced fibrosis; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblast (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 5; (e). qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by TGF-β (n = 8, *P < 0.01 vs. control).
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC5120308&req=5

f3: TGF-β-induced fibrosis and mediated increase in serpinE2 expression.(a) qRT-PCR assay was applied for detection of collagen I and collagen III mRNA expression in myocardial fibroblast (n = 6, *P < 0.05 vs. control) in TGF-β-induced fibrosis; (b) Sircol™ Collagen Assay was used to quantify the total collagen concentration both in fibroblasts and in the supernatants of fibroblasts (n = 6, *P < 0.05 vs. control); (c) The expression of serpinE2 was detected by ELISA in the supernatants of fibroblast (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 5; (e). qRT-PCR was also used to detect expression of serpinE2 mRNA in myocardial fibroblasts stimulated by TGF-β (n = 8, *P < 0.01 vs. control).
Mentions: It has well been accepted that the activation of renin-angiotensin-aldosterone system would increase levels of active TGF-β that plays an important function in pressure overload-induced cardiac fibrosis20 and lead to increase in the mRNA expressions of collagen I and III in myocardial fibroblasts2021. Compared with the control group, the total collagen concentration detected by Sircol™ Collagen Assay were significantly elevated in myocardial fibroblasts after 24 h and 48 h stimulation with TGF-β 10 ng/ml (Fig. 3a and b), meanwhile, quantified serpinE2 secretion by ELISA assay into the supernatants of fibroblast culture medium showed accordant changes with collagen content. These results indicate that TGF-β also promotes serpinE2 secretion from fibroblasts into the supernatants (Fig. 3c). The supportive results were accordingly also obtained using western blot and qRT-PCR showing the up-regulation of both protein and mRNA expression of serpinE2 in myocardial fibroblasts treated with TGF-β (Fig. 3d 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