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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 Ang II-induced MCF proliferation. a, MCF proliferation was assayed with CCK-8. b-c, PCNA expression was measured by western blot (b) and quantified by normalizing to α-Tubulin (c). *P < 0.01 vs. vehicle-treated cells (−), #P < 0.01 vs. Ang II-treated cells, &P < 0.05 vs. SOD1-treated cells (n = 5)
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Fig3: Effect of PEP-1-SOD1 on Ang II-induced MCF proliferation. a, MCF proliferation was assayed with CCK-8. b-c, PCNA expression was measured by western blot (b) and quantified by normalizing to α-Tubulin (c). *P < 0.01 vs. vehicle-treated cells (−), #P < 0.01 vs. Ang II-treated cells, &P < 0.05 vs. SOD1-treated cells (n = 5)

Mentions: Ang II is a potent factor inducing MCF proliferation. We confirmed that Ang II promotes MCF proliferation (Fig. 3a). Pretreatment of SOD1 slightly, but transduction of PEP-1-SOD1 fusion proteins dramatically attenuated the Ang II-induced cell proliferation. Proliferating cell nuclear antigen (PCNA) is a biomarker of cell proliferation. To further determine the effect of PEP-1-SOD1 fusion protein on MCF proliferation, we detected the PCNA expression. Ang II up-regulated PCNA expression (Fig. 3b-c). SOD1 transduction, slightly inhibited, but PEP-1-SOD1 fusion proteins dramatically blocked PCNA expression (Fig. 3b-c). These results indicate that Ang II induces MCF proliferation through increasing ROS production, which can be diminished by the transduction of PEP-1-SOD1.Fig. 3


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 Ang II-induced MCF proliferation. a, MCF proliferation was assayed with CCK-8. b-c, PCNA expression was measured by western blot (b) and quantified by normalizing to α-Tubulin (c). *P < 0.01 vs. vehicle-treated cells (−), #P < 0.01 vs. Ang II-treated cells, &P < 0.05 vs. SOD1-treated cells (n = 5)
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

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

Fig3: Effect of PEP-1-SOD1 on Ang II-induced MCF proliferation. a, MCF proliferation was assayed with CCK-8. b-c, PCNA expression was measured by western blot (b) and quantified by normalizing to α-Tubulin (c). *P < 0.01 vs. vehicle-treated cells (−), #P < 0.01 vs. Ang II-treated cells, &P < 0.05 vs. SOD1-treated cells (n = 5)
Mentions: Ang II is a potent factor inducing MCF proliferation. We confirmed that Ang II promotes MCF proliferation (Fig. 3a). Pretreatment of SOD1 slightly, but transduction of PEP-1-SOD1 fusion proteins dramatically attenuated the Ang II-induced cell proliferation. Proliferating cell nuclear antigen (PCNA) is a biomarker of cell proliferation. To further determine the effect of PEP-1-SOD1 fusion protein on MCF proliferation, we detected the PCNA expression. Ang II up-regulated PCNA expression (Fig. 3b-c). SOD1 transduction, slightly inhibited, but PEP-1-SOD1 fusion proteins dramatically blocked PCNA expression (Fig. 3b-c). These results indicate that Ang II induces MCF proliferation through increasing ROS production, which can be diminished by the transduction of PEP-1-SOD1.Fig. 3

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