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Reactive oxygen species effect PASMCs apoptosis via regulation of dynamin-related protein 1 in hypoxic pulmonary hypertension

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Abstract

The high level of reactive oxygen species and up-regulation of mitochondrial fission protein dynamin-related protein-1, both of which involved in pulmonary artery smooth muscle cells (PASMCs) apoptosis, have been detected in the lungs of rodent pulmonary arterial hypertension models. However, the regulatory mechanisms between ROS and DRP1 are poorly understood. In this study, ROS inhibitor, hypoxic rodent PAH models, small interfering RNA, polymerase chain reaction, Western blot, flow cytometry, immunohistochemistry and immunofluorescence were used. We determined that ROS, mainly derive from mitochondria, mediate mitochondria fission of PASMCs contributing to pulmonary vascular remodeling. Meanwhile, we also observed that hypoxia-induced DRP1 expression depends on ROS generation, especially mitochondrial ROS (mROS). Moreover, the levels of ROS and mROS evoked by hypoxia were regulated by DRP1. Furthermore, we verified the apoptosis suppression of PASMCs under hypoxia due to the interaction between ROS/mROS and DRP1. Our study reveals a novel mechanism of hypoxia-induced pulmonary vascular remodeling, suggesting a new therapeutic strategy which is targeting on the positive feedback of ROS/mROS-DRP1 for the treatment of PAH.

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Abbreviations

DCFDA:

2′,7′-Dichlorofluorescin diacetate

DRP1:

Dynamin-related protein-1

MFC:

Mitochondrial fragmentation count (MFC)

NAC:

N-acetylcysteine

PASMCs:

Pulmonary artery smooth muscle cells

PAH:

Pulmonary arterial hypertension

PVR:

Pulmonary vascular remodeling

ROS:

Reactive oxygen species

TEMPO/MitoTEMPO:

2-(2,2,6,6-Tetramethylpiperidin-1-oxyl-4-ylamino)-2-oxoethyl)triphenylphosphonium chloride

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Acknowledgments

This work was supported by these fundings: Contract Grant Sponsor: the Graduate Innovation Foundation of Heilongjiang Province; Contract Grant Number: YJSCX 2014-15HYD; Contract Grant Sponsor: Natural Science Foundation of China; Contract Grant Numbers: 31471095, 81270113, 81400353; Contract Grant Sponsor: Key Research Plan of National Natural Science Foundation of China; Contract Grant Number: 91339107; Contract Grant Sponsor: Natural Science Foundation of Heilongjiang Province; Contract Grant Numbers: SCX-2012-9, QC2014C096; Contract Grant Sponsor: Wu Liande Youth Science Foundation; Contract Grant Number: WLD-QN1410; Contract Grant Sponsor: Postdoctoral Foundation of Heilongjiang Province, China; Contract Grant Numbers: LBH-Z14133, 2015m571438.

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Authors and Affiliations

  1. Department of Biopharmaceutical Sciences, College of Pharmacy, Harbin Medical University (Daqing), Daqing, 163319, People’s Republic of China

    Lixin Zhang, Chen Zhang, Mingfei Ma, Fengying Zhang, Linlin Zhang & Daling Zhu

  2. Biopharmaceutical Key Laboratory of Heilongjiang Province, Harbin, 150081, People’s Republic of China

    Lixin Zhang, Chen Zhang, Mingfei Ma, Fengying Zhang, Linlin Zhang, Shuzhen Li & Daling Zhu

  3. Department of Immunology, College of Medical Laboratory Science and Technology, Harbin Medical University (Daqing), Daqing, 163319, People’s Republic of China

    Cui Ma & Yingli Chen

  4. Department of Physiology, College of Basic Medicine, Harbin Medical University (Daqing), Daqing, 163319, People’s Republic of China

    Fangyuan Cao

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  1. Lixin Zhang
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  2. Cui Ma
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Correspondence to Daling Zhu.

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Zhang, L., Ma, C., Zhang, C. et al. Reactive oxygen species effect PASMCs apoptosis via regulation of dynamin-related protein 1 in hypoxic pulmonary hypertension. Histochem Cell Biol 146, 71–84 (2016). https://doi.org/10.1007/s00418-016-1424-9

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