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20 July 2019 : Laboratory Research  

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

Yu Yang123ABCDEF, Shiping Chen123ABCDE, Ling Tao3ABCD, Shiquan Gan23ABCD, Hong Luo23ACD, Yini Xu23ACD, Xiangchun Shen123ABCDFG*

DOI: 10.12659/MSM.915542

Med Sci Monit 2019; 25:5375-5388

Abstract

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 AND 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.

Keywords: Aldosterone, Fibroblasts, NF-E2-Related Factor 2, Reactive Oxygen Species, Alkaloids, Animals, Newborn, Cell Differentiation, Cell Transdifferentiation, Collagen, Fibrosis, Heart, Kelch-Like ECH-Associated Protein 1, Myocardium, myofibroblasts, Quinolizines

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750