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Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro

BACKGROUND: Oxymatrine (OMT), a quinolizidine alkaloid derived from the traditional Chinese herb Radix Sophorae flavescentis, has widely reported pharmacological efficacy in treating cardiovascular dysfunction-related diseases. However, the underlying mechanism has been unclear. Here, we investigate...

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Autores principales: Yang, Yu, Chen, Shiping, Tao, Ling, Gan, Shiquan, Luo, Hong, Xu, Yini, Shen, Xiangchun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: International Scientific Literature, Inc. 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6662943/
https://www.ncbi.nlm.nih.gov/pubmed/31325292
http://dx.doi.org/10.12659/MSM.915542
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author Yang, Yu
Chen, Shiping
Tao, Ling
Gan, Shiquan
Luo, Hong
Xu, Yini
Shen, Xiangchun
author_facet Yang, Yu
Chen, Shiping
Tao, Ling
Gan, Shiquan
Luo, Hong
Xu, Yini
Shen, Xiangchun
author_sort Yang, Yu
collection PubMed
description BACKGROUND: Oxymatrine (OMT), a quinolizidine alkaloid derived from the traditional Chinese herb Radix Sophorae flavescentis, has widely reported pharmacological efficacy in treating cardiovascular dysfunction-related diseases. However, the underlying mechanism has been unclear. Here, we investigated the potential inhibitory effects and mechanism of OMT on transdifferentiation of cardiac fibroblast to myofibroblasts induced by aldosterone in vitro. MATERIAL/METHODS: The cardiac fibroblasts (CFBs) proliferation and migration capacity were evaluated by MTT assay, cell cycle assay, and scratch analysis, respectively. The protein expression of the Nrf2/Keap1 signal pathway, FN, Collagen I, Collagen III, α-SMA, CTGF, and mineralocorticoid receptor (MR) protein was detected by Western blot analysis. The mRNA expression of Nrf2 was detected by qRT-PCR. Immunofluorescence staining was used to observe the expression of α-SMA protein. Nrf2 siRNA was used to explore the role of Nrf2 in OMT-treated CFBs. GSH, SOD, and MDA levels and hydroxyproline content were measured by colorimetric assay with commercial kits. The DCFH-DA fluorescent probe was used to assess cellular ROS levels. RESULTS: OMT and Curcumin (an Nrf2 agonist) attenuated aldosterone (ALD)-induced proliferation and migration in CFBs, as well as the fibrosis-associated protein expression levels. Moreover, OMT activated Nrf2 and promoted the nucleus translocation of Nrf2. OMT alleviated the elevated levels of α-SMA, Collagen I, Collagen III, and CTGF, which were abrogated by the Nrf2 siRNA transfection. We also found that OMT decreased oxidative stress levels. CONCLUSIONS: Our results confirm that OMT alleviates transdifferentiation of cardiac fibroblasts to myofibroblasts induced by aldosterone via activating the Nrf2/Keap1 pathway in vitro.
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spelling pubmed-66629432019-08-16 Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro Yang, Yu Chen, Shiping Tao, Ling Gan, Shiquan Luo, Hong Xu, Yini Shen, Xiangchun Med Sci Monit Lab/In Vitro Research BACKGROUND: Oxymatrine (OMT), a quinolizidine alkaloid derived from the traditional Chinese herb Radix Sophorae flavescentis, has widely reported pharmacological efficacy in treating cardiovascular dysfunction-related diseases. However, the underlying mechanism has been unclear. Here, we investigated the potential inhibitory effects and mechanism of OMT on transdifferentiation of cardiac fibroblast to myofibroblasts induced by aldosterone in vitro. MATERIAL/METHODS: The cardiac fibroblasts (CFBs) proliferation and migration capacity were evaluated by MTT assay, cell cycle assay, and scratch analysis, respectively. The protein expression of the Nrf2/Keap1 signal pathway, FN, Collagen I, Collagen III, α-SMA, CTGF, and mineralocorticoid receptor (MR) protein was detected by Western blot analysis. The mRNA expression of Nrf2 was detected by qRT-PCR. Immunofluorescence staining was used to observe the expression of α-SMA protein. Nrf2 siRNA was used to explore the role of Nrf2 in OMT-treated CFBs. GSH, SOD, and MDA levels and hydroxyproline content were measured by colorimetric assay with commercial kits. The DCFH-DA fluorescent probe was used to assess cellular ROS levels. RESULTS: OMT and Curcumin (an Nrf2 agonist) attenuated aldosterone (ALD)-induced proliferation and migration in CFBs, as well as the fibrosis-associated protein expression levels. Moreover, OMT activated Nrf2 and promoted the nucleus translocation of Nrf2. OMT alleviated the elevated levels of α-SMA, Collagen I, Collagen III, and CTGF, which were abrogated by the Nrf2 siRNA transfection. We also found that OMT decreased oxidative stress levels. CONCLUSIONS: Our results confirm that OMT alleviates transdifferentiation of cardiac fibroblasts to myofibroblasts induced by aldosterone via activating the Nrf2/Keap1 pathway in vitro. International Scientific Literature, Inc. 2019-07-20 /pmc/articles/PMC6662943/ /pubmed/31325292 http://dx.doi.org/10.12659/MSM.915542 Text en © Med Sci Monit, 2019 This work is licensed under Creative Common Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/) )
spellingShingle Lab/In Vitro Research
Yang, Yu
Chen, Shiping
Tao, Ling
Gan, Shiquan
Luo, Hong
Xu, Yini
Shen, Xiangchun
Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro
title Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro
title_full Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro
title_fullStr Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro
title_full_unstemmed Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro
title_short Inhibitory Effects of Oxymatrine on Transdifferentiation of Neonatal Rat Cardiac Fibroblasts to Myofibroblasts Induced by Aldosterone via Keap1/Nrf2 Signaling Pathways In Vitro
title_sort inhibitory effects of oxymatrine on transdifferentiation of neonatal rat cardiac fibroblasts to myofibroblasts induced by aldosterone via keap1/nrf2 signaling pathways in vitro
topic Lab/In Vitro Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6662943/
https://www.ncbi.nlm.nih.gov/pubmed/31325292
http://dx.doi.org/10.12659/MSM.915542
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