title: Downregulation of cystathionine β-synthase/hydrogen sulfide contributes to rotenone-induced microglia polarization toward M1 type.
abstract: Microglia-mediated neuroinflammation is implicated in the pathogenesis of several neurodegenerative disorders. Microglia can be activated and polarized to exert pro- or anti-inflammatory roles in response to specific stimulus. Rotenone is an environmental toxin that has been shown to activate microglia and neuroinflammation. However, the effects and mechanisms of rotenone on microglia polarization are poorly studied. In the present study, we demonstrated that rotenone enhanced the levels of M1 phenotypic genes including TNF-α, iNOS and COX-2/PGE2 but reduced that of M2 markers such as Ym1/2 and IL-10 in mouse primary and immortalized microglia. Moreover, the transcription and protein expression of cystathionine-β-synthase (CBS), as well as hydrogen sulfide (H2S) production were decreased in rotenone-treated primary microglia. Elevating endogenous H2S via CBS over-expression in immortalized microglia not only reduced the expression of pro-inflammatory M1 genes, but also enhanced the anti-inflammatory M2 marker IL-10 production in response to rotenone stimulation as compared to vector-transfected cells. Similarly, pretreatment with H2S donor NaHS (50, 100 and 500μmol/L) attenuated the increases of M1 gene expression triggered by rotenone treatment, and enhanced the M2 gene Ym1/2 expression in mouse primary microglia. In addition, we observed reactive oxygen species (ROS) scavenger N-acetyl-l-cysteine reversed the down-regulation of CBS and H2S generation caused by rotenone in microglia. NaHS pretreatment also decreased the ROS formation in rotenone-stimulated microglia. Taken together, these results reveal that probably via triggering ROS formation, rotenone suppressed the CBS-H2S pathway and thus promoted microglia polarization toward M1 pro-inflammatory phenotype.
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China; Department of Pharmacology, School of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
authors.forename: Chenchen
authors.initials: C
authors.lastname: Du
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China; Department of Neurology, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
authors.forename: Mengmeng
authors.initials: M
authors.lastname: Jin
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China; Department of Neurology, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Hong
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China; Department of Pharmacology, School of Pharmaceutical Science, Soochow University, Suzhou 215123, China.
authors.forename: Qian
authors.initials: Q
authors.lastname: Li
authors.affiliation: Department of Neurology, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
authors.forename: Xian-Hui
authors.initials: XH
authors.lastname: Wang
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China.
authors.forename: Jin-Min
authors.initials: JM
authors.lastname: Xu
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China.
authors.forename: Fen
authors.initials: F
authors.lastname: Wang
authors.affiliation: Cytoneurobiology Unit & Department of Anatomy, Medical College of Soochow University, Suzhou 215123, China.
authors.forename: Ye
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Jia
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China; Department of Neurology, The Second Affiliated Hospital of Soochow University, Suzhou 215004, China.
authors.forename: Chun-Feng
authors.initials: CF
authors.lastname: Liu
authors.affiliation: Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and Institute of Neuroscience, Soochow University, Suzhou, Jiangsu 215021, China; Department of Pharmacology, School of Pharmaceutical Science, Soochow University, Suzhou 215123, China. Electronic address: hulifang@suda.edu.cn.
authors.forename: Li-Fang
authors.initials: LF
authors.lastname: Hu
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D003541:Cystathionine beta-Synthase / Q000235:genetics* / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects; D006862:Hydrogen Sulfide / Q000378:metabolism*; D018836:Inflammation Mediators / Q000378:metabolism; D016753:Interleukin-10 / Q000096:biosynthesis; D051379:Mice; D017628:Microglia / Q000145:classification / Q000187:drug effects* / Q000378:metabolism*; D018696:Neuroprotective Agents / Q000378:metabolism; D009498:Neurotoxins / Q000633:toxicity; D010641:Phenotype; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D012402:Rotenone / Q000633:toxicity*
chemical_list: C556062:IL10 protein, mouse; D018836:Inflammation Mediators; D018696:Neuroprotective Agents; D009498:Neurotoxins; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; D012402:Rotenone; D016753:Interleukin-10; D003541:Cystathionine beta-Synthase; D006862:Hydrogen Sulfide
keywords: Cystathionine β-synthase; Hydrogen sulfide; Microglia; Neuroinflammation; Polarization; Rotenone