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PEP-1-SOD1 fusion proteins block cardiac myofibroblast activation and angiotensin II-induced collagen production.

Tan LG, Xiao JH, Yu DL, Zhang L, Zheng F, Guo LY, Yang JY, Tang JM, Chen SY, Wang JN - BMC Cardiovasc Disord (2015)

Bottom Line: However, the effect of copper, zinc-superoxide dismutase (SOD1) as a therapeutic agent is limited due to the insufficient transduction.Protein expression was analyzed by western blotting.PEP-1-SOD1 suppressed MCF proliferation and differentiation and reduced production of collagen type I and III.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Medicine, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, P. R. China. 28621855@qq.com.

ABSTRACT

Background: Oxidative stress is closely associated with cardiac fibrosis. However, the effect of copper, zinc-superoxide dismutase (SOD1) as a therapeutic agent is limited due to the insufficient transduction. This study was aimed to investigate the effect of PEP-1-SOD1 fusion protein on angiotensin II (ANG II)-induced collagen metabolism in rat cardiac myofibroblasts (MCFs).

Methods: MCFs were pretreated with SOD1 or PEP-1-SOD1 fusion protein for 2 h followed by incubation with ANG II for 24 h. Cell proliferation was measured by Cell Counting Kit-8. Superoxide anion productions were detected by both fluorescent microscopy and Flow Cytometry. MMP-1 and TIMP-1 were determined by ELISA. Intracellular MDA content and SOD activity were examined by commercial assay kits. Protein expression was analyzed by western blotting.

Results: PEP-1-SOD1 fusion protein efficiently transduced into MCF, scavenged intracellular O2 (-), decreased intracellular MDA content, increased SOD activity, suppressed ANG II-induced proliferation, reduced expression of TGF-β1, α-SMA, collagen type I and III, restored MMP-1 secretion, and attenuated TIMP-1 secretion.

Conclusion: PEP-1-SOD1 suppressed MCF proliferation and differentiation and reduced production of collagen type I and III. Therefore, PEP-1-SOD1 fusion protein may be a potential novel therapeutic agent for cardiac fibrosis.

No MeSH data available.


Related in: MedlinePlus

Effect of PEP-1-SOD1 on α-SMA and TGF-β1 protein expression. a α-SMA and TGF-β1 protein expression was detected by western blot. b Quantification of α-SMA expression by normalizing to α-Tubulin. c Quantification of TGF-β1 expression by normalizing to α-Tubulin. *P < 0.01 vs. vehicle-treated cells, #P < 0.01 vs. Ang II-treated cells. &P < 0.01 vs. SOD1-transduced cells n = 5
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Fig4: Effect of PEP-1-SOD1 on α-SMA and TGF-β1 protein expression. a α-SMA and TGF-β1 protein expression was detected by western blot. b Quantification of α-SMA expression by normalizing to α-Tubulin. c Quantification of TGF-β1 expression by normalizing to α-Tubulin. *P < 0.01 vs. vehicle-treated cells, #P < 0.01 vs. Ang II-treated cells. &P < 0.01 vs. SOD1-transduced cells n = 5

Mentions: Excessive MCF activation accompanied by excessive collagen production is the main mechanism underlying the onset of cardiac fibrosis. Ang II markedly stimulated expression of smooth muscle α-actin (α-SMA) and TGF-β1 in MCF (Fig. 4a-c), indicating a MCF activation. It is known that Ang II induces fibrosis through TGF-β signaling pathway. SOD1 slightly reduced Ang II-induced α-SMA and TGF-β expression. PEP-1-SOD1 infusion protein, however, dramatically blocked Ang II induction of these two genes (Fig. 4). Importantly, PEP-1-SOD1 had a much greater effect as compared to SOD1 (Fig. 4).Fig. 4


PEP-1-SOD1 fusion proteins block cardiac myofibroblast activation and angiotensin II-induced collagen production.

Tan LG, Xiao JH, Yu DL, Zhang L, Zheng F, Guo LY, Yang JY, Tang JM, Chen SY, Wang JN - BMC Cardiovasc Disord (2015)

Effect of PEP-1-SOD1 on α-SMA and TGF-β1 protein expression. a α-SMA and TGF-β1 protein expression was detected by western blot. b Quantification of α-SMA expression by normalizing to α-Tubulin. c Quantification of TGF-β1 expression by normalizing to α-Tubulin. *P < 0.01 vs. vehicle-treated cells, #P < 0.01 vs. Ang II-treated cells. &P < 0.01 vs. SOD1-transduced cells n = 5
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4597385&req=5

Fig4: Effect of PEP-1-SOD1 on α-SMA and TGF-β1 protein expression. a α-SMA and TGF-β1 protein expression was detected by western blot. b Quantification of α-SMA expression by normalizing to α-Tubulin. c Quantification of TGF-β1 expression by normalizing to α-Tubulin. *P < 0.01 vs. vehicle-treated cells, #P < 0.01 vs. Ang II-treated cells. &P < 0.01 vs. SOD1-transduced cells n = 5
Mentions: Excessive MCF activation accompanied by excessive collagen production is the main mechanism underlying the onset of cardiac fibrosis. Ang II markedly stimulated expression of smooth muscle α-actin (α-SMA) and TGF-β1 in MCF (Fig. 4a-c), indicating a MCF activation. It is known that Ang II induces fibrosis through TGF-β signaling pathway. SOD1 slightly reduced Ang II-induced α-SMA and TGF-β expression. PEP-1-SOD1 infusion protein, however, dramatically blocked Ang II induction of these two genes (Fig. 4). Importantly, PEP-1-SOD1 had a much greater effect as compared to SOD1 (Fig. 4).Fig. 4

Bottom Line: However, the effect of copper, zinc-superoxide dismutase (SOD1) as a therapeutic agent is limited due to the insufficient transduction.Protein expression was analyzed by western blotting.PEP-1-SOD1 suppressed MCF proliferation and differentiation and reduced production of collagen type I and III.

View Article: PubMed Central - PubMed

Affiliation: Institute of Clinical Medicine, Renmin Hospital, Hubei University of Medicine, Shiyan, Hubei, 442000, P. R. China. 28621855@qq.com.

ABSTRACT

Background: Oxidative stress is closely associated with cardiac fibrosis. However, the effect of copper, zinc-superoxide dismutase (SOD1) as a therapeutic agent is limited due to the insufficient transduction. This study was aimed to investigate the effect of PEP-1-SOD1 fusion protein on angiotensin II (ANG II)-induced collagen metabolism in rat cardiac myofibroblasts (MCFs).

Methods: MCFs were pretreated with SOD1 or PEP-1-SOD1 fusion protein for 2 h followed by incubation with ANG II for 24 h. Cell proliferation was measured by Cell Counting Kit-8. Superoxide anion productions were detected by both fluorescent microscopy and Flow Cytometry. MMP-1 and TIMP-1 were determined by ELISA. Intracellular MDA content and SOD activity were examined by commercial assay kits. Protein expression was analyzed by western blotting.

Results: PEP-1-SOD1 fusion protein efficiently transduced into MCF, scavenged intracellular O2 (-), decreased intracellular MDA content, increased SOD activity, suppressed ANG II-induced proliferation, reduced expression of TGF-β1, α-SMA, collagen type I and III, restored MMP-1 secretion, and attenuated TIMP-1 secretion.

Conclusion: PEP-1-SOD1 suppressed MCF proliferation and differentiation and reduced production of collagen type I and III. Therefore, PEP-1-SOD1 fusion protein may be a potential novel therapeutic agent for cardiac fibrosis.

No MeSH data available.


Related in: MedlinePlus