title: Mesenchymal stem cells-conditioned medium protects PC12 cells against 2,5-hexanedione-induced apoptosis via inhibiting mitochondria-dependent caspase 3 pathway.
abstract: Studies suggested that the conditioned medium of mesenchymal stem cells (MSC-CM) inhibited the increased apoptosis in various cells. However, there are no reports underlying the protection of MSC-CM against 2,5-hexanedione (HD)-induced apoptosis in neural cells. In the present study, the viability was observed in PC12 cells that received HD alone or with MSC-CM by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Apoptosis was estimated by Hoechst 33342 staining and flow cytometry. Mitochondrial transmembrane potential was examined by rhodamine 123. Moreover, we investigated the expression of Bax and Bcl-2, cytochrome c translocation, and caspase 3 activity by real-time polymerase chain reaction, Western blot, and immunochemistry. Nerve growth factor (NGF) was examined in MSCs and MSC-CM. Our results showed that MSC-CM promoted cell survival and reduced apoptosis in HD-exposed PC12 cells. Moreover, MSC-CM significantly reversed disturbance of Bax and Bcl-2, ameliorated disruption of mitochondrial transmembrane potential, and reduced release of cytochrome c and activity of caspase 3 in HD-exposed PC12 cells. In the meantime, NGF was detected in MSCs and MSC-CM. These findings demonstrate that MSC-CM protects against HD-induced apoptosis in PC12 cells via inhibiting mitochondrial pathway. Our results indicate that NGF in MSC-CM may be involved in the protection of MSC-CM against HD-induced apoptosis. Our study clarifies the protection of MSC-CM on HD neurotoxicity and its underlying mechanism.
authors.affiliation: 1 Department of Occupational and Environmental Health, Dalian Medical University, Dalian, Liaoning, China.
authors.forename: Shuang-Yue
authors.initials: SY
authors.lastname: Li
authors.affiliation: 1 Department of Occupational and Environmental Health, Dalian Medical University, Dalian, Liaoning, China.
authors.forename: Yuan
authors.initials: Y
authors.lastname: Qi
authors.affiliation: 2 College of Stomatology, Dalian Medical University, Dalian, Liaoning, China.
authors.forename: Shu-Hai
authors.initials: SH
authors.lastname: Hu
authors.affiliation: 1 Department of Occupational and Environmental Health, Dalian Medical University, Dalian, Liaoning, China.
authors.forename: Feng-Yuan
authors.initials: FY
authors.lastname: Piao
authors.affiliation: 3 Department of Obstetrics and Gynecology, Dalian, Liaoning, China.
authors.forename: Huai
authors.initials: H
authors.lastname: Guan
authors.affiliation: 1 Department of Occupational and Environmental Health, Dalian Medical University, Dalian, Liaoning, China.
authors.forename: Zhe-Min
authors.initials: ZM
authors.lastname: Wang
authors.affiliation: 1 Department of Occupational and Environmental Health, Dalian Medical University, Dalian, Liaoning, China.
authors.forename: Ruo-Lin
authors.initials: RL
authors.lastname: Chen
authors.affiliation: 1 Department of Occupational and Environmental Health, Dalian Medical University, Dalian, Liaoning, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Liu
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3 / Q000187:drug effects / Q000502:physiology*; D005434:Flow Cytometry; D006588:Hexanones / Q000494:pharmacology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects / Q000502:physiology; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D016716:PC12 Cells / Q000187:drug effects*; D051381:Rats; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction / Q000187:drug effects* / Q000502:physiology
chemical_list: D006588:Hexanones; C011269:2,5-hexanedione; D053148:Caspase 3
keywords: 2, 5-Hexanedione; PC12 cells; apoptosis; conditioned medium of mesenchymal stem cells; mesenchymal stem cells; mitochondria-dependent caspase 3 pathway