title: Modification of HSP proteins and Ca2+ are responsible for the NO-derived peroxynitrite mediated neurological damage in PC12 cell.
abstract: Peroxynitrite as one crucial metabolite of NO-derived agents has been well multi-investigated to inspect its potential role and sought to define its concrete mechanism underlying the memory loss and impaired cognition involved in pathological processes. In this investigation, the cell viability was assessed by the MTT assay. The neurotoxicity of peroxynitrite was analyzed by using immunohistochemical measurements in cultured PC12 cells to explore the underlying mechanisms. The generation of ROS was evaluated by a fluorometry assay by a fluorometry assay. Apoptosis was assayed by annexin V-FITC and PI staining with flow cytometry. [Ca2+]i was examined by using the microspectrofluorometer. Hsp70 was detected by western blot assay. The results revealed that PC12 cells were inhibited by peroxynitrite both in a dose-dependent and time-dependent manner. The level of ROS in PC12 cells exposed to SIN-1 was increased in a dose-dependent manner. The result indicated that the SIN-1 induced apoptosis of PC12 cells in a dose-dependent manner. Quercetin inhibited the viability of PC12 cells in a concentration-dependent manner. [Ca2+]i was increased gradually when cells treated with quercetin alone and also increased with treatment of dantrolene-containing. Hsp70 was significantly decreased in SIN-1-treated group compared with that of control group (P<0.01). In conclusion, Ca2+ homeostasis and chaperone Hsp70 were critically involved in peroxynitrite induced nitrosative stress as protective. Peroxynitrite acts as the pathological agent in learning and memory defects in CNS disorders associated with challenge.
authors.affiliation: Department of Emergency, Xi'an Children's Hospital Xi'an 710003, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Wen
authors.affiliation: Department of Neurology, Xi'an Children's Hospital Xi'an 710003, China.
authors.forename: Hua
authors.initials: H
authors.lastname: Li
authors.affiliation: Department of Neurology, Xi'an Children's Hospital Xi'an 710003, China.
authors.forename: Yudan
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Department of Neurology, Xi'an Children's Hospital Xi'an 710003, China.
authors.forename: Xia
authors.initials: X
authors.lastname: Li
authors.affiliation: Department of Ultrasound, Xi'an Children's Hospital Xi'an 710003, China.
authors.forename: Fang
authors.initials: F
authors.lastname: Liu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015153:Blotting, Western; D002118:Calcium / Q000378:metabolism*; D002470:Cell Survival / Q000502:physiology; D005434:Flow Cytometry; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D007150:Immunohistochemistry; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D009569:Nitric Oxide / Q000378:metabolism*; D016716:PC12 Cells; D030421:Peroxynitrous Acid / Q000633:toxicity*; D051381:Rats; D017382:Reactive Oxygen Species / Q000633:toxicity*
chemical_list: D018840:HSP70 Heat-Shock Proteins; D017382:Reactive Oxygen Species; D030421:Peroxynitrous Acid; D009569:Nitric Oxide; D002118:Calcium
keywords: Ca2+; Hsp70; Peroxynitrite; cognition; neurotoxicity