title: Angiotensin AT2 receptor stimulation inhibits activation of NADPH oxidase and ameliorates oxidative stress in rotenone model of Parkinson's disease in CATH.a cells.
abstract: Oxidative stress has long been considered as a major contributing factor in the pathogenesis of Parkinson's disease (PD). The brain has an independent local renin-angiotensin system (RAS). Angiotensin II (Ang II) activates NADPH-dependent oxidases, which are a major source of superoxide and are upregulated in major aging-related diseases such as hypertension and neurodegenerative disease. In this study, we firstly examined whether CGP42112, an AT2 receptor (AT2R) agonist, may exert direct protective effects on the rotenone-induced CATH.a cell injury in vitro. We used CATH.a cell line to evaluate changes in cultured dopaminergic neuron levels of superoxide dismutase (SOD), glutathione (GSH) and reactive oxygen species (ROS). We also evaluated expression of NADPH oxidase, AT1 and AT2 receptors in treated with phosphate buffer saline (PBS), rotenone, Ang II, AT2R agonist CGP42112, or AT2R antagonist PD123319, alone and combined (n=6, each group). Quantitative reverse transcriptase PCR (qRT-PCR) and western blot were used to determine messenger RNA (mRNA) and protein levels of the AT1, AT2 receptors and NADPH oxidase. ROS generation was determined by the dichlorodihydrofluorescein diacetate fluorescent probe assay. The levels of SOD and GSH were measured by using available kits. In our study, CGP42112 (100nM) significantly reduced rotenone-induced oxidative stress and elevated the total SOD activity and GSH level. In addition, CGP42112 significantly increased AT2R expression and attenuated Ang II-induced NADPH oxidase activation, and these effects were completely abolished by the AT2R antagonist, PD123319 (1μM). Our results suggest that CGP42112 attenuates rotenone-induced oxidative stress in CATH.a neuron via activating AT2R and suppressing NADPH oxidase expression.
authors.affiliation: Department of Neurology, Nanjing Brain Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, PR China.
authors.forename: Jie
authors.initials: J
authors.lastname: Lu
authors.affiliation: Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu 210006, PR China.
authors.forename: Liang
authors.initials: L
authors.lastname: Wu
authors.affiliation: Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu 210006, PR China.
authors.forename: Teng
authors.initials: T
authors.lastname: Jiang
authors.affiliation: Department of Neurology, Nanjing Brain Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, PR China.
authors.forename: Yao
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu 210006, PR China.
authors.forename: Hongrui
authors.initials: H
authors.lastname: Zhao
authors.affiliation: Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu 210006, PR China.
authors.forename: Qing
authors.initials: Q
authors.lastname: Gao
authors.affiliation: Department of Neurology, Nanjing Brain Hospital, Nanjing Medical University, Nanjing, Jiangsu 210029, PR China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Pan
authors.affiliation: Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu 210006, PR China.
authors.forename: Youyong
authors.initials: Y
authors.lastname: Tian
authors.affiliation: Department of Neurology, Nanjing First Hospital, Nanjing Medical University, Nanjing, Jiangsu 210006, PR China. Electronic address: zhangyingdong@aliyun.com.
authors.forename: Yingdong
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000704:Analysis of Variance; D057912:Angiotensin II Type 2 Receptor Blockers / Q000494:pharmacology; D000809:Angiotensins / Q000494:pharmacology; D000818:Animals; D045744:Cell Line, Tumor; D005786:Gene Expression Regulation / Q000187:drug effects; D005978:Glutathione / Q000378:metabolism; D007093:Imidazoles / Q000494:pharmacology; D007306:Insecticides / Q000494:pharmacology*; D051379:Mice; D019255:NADPH Oxidases / Q000378:metabolism*; D009842:Oligopeptides / Q000494:pharmacology; D018384:Oxidative Stress / Q000187:drug effects*; D011725:Pyridines / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D044140:Receptor, Angiotensin, Type 1 / Q000235:genetics / Q000378:metabolism; D044139:Receptor, Angiotensin, Type 2 / Q000235:genetics / Q000378:metabolism*; D012402:Rotenone / Q000494:pharmacology*; D013482:Superoxide Dismutase / Q000378:metabolism
chemical_list: D057912:Angiotensin II Type 2 Receptor Blockers; D000809:Angiotensins; D007093:Imidazoles; D007306:Insecticides; D009842:Oligopeptides; D011725:Pyridines; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D044140:Receptor, Angiotensin, Type 1; D044139:Receptor, Angiotensin, Type 2; D012402:Rotenone; C060894:CGP 42112A; C073402:PD 123319; D013482:Superoxide Dismutase; D019255:NADPH Oxidases; D005978:Glutathione
keywords: CATH.a neurons; CGP42112; Oxidative stress; Parkinson's disease; Rotenone