title: β-Asarone protects PC12 cells against OGD/R-induced injury via attenuating Beclin-1-dependent autophagy.
abstract: OBJECTIVE
  To explore the effects of β-asarone from Acorus Tatarinowii Schott on autophagy in an ischemic stroke model of PC12 cells.
  
  
  METHODS
  The ischemic stroke model of PC12 cells was made by OGD/R (2 h oxygen-glucose deprivation followed by 24 h reperfusion). Drug administration was started 1 h before OGD and last for 3 h. Then the cells were incubated in the drug-free and full culture medium under normoxic conditions for 24 h. After the treatments, Beclin-1, intracellular free calcium concentration ([Ca(2+)](i)) and mitochondrial membrane potential (MMP) were analyzed using flow cytometry. Cell viability was measured using MTT assay. Cell morphology was studied under inverted phase contrast microscope, and autophagosomes were observed under transmission electron microscope.
  
  
  RESULTS
  Pretreatment with β-asarone (20, 30, or 45 μg/mL) or the calcium channel antagonist nimodipine (10 μmol/L) significantly increased the cell viability and MMP, and decreased Beclin-1 expression and [Ca(2+)](i) in OGD/R-treated PC12 cells. Under inverted phase contrast microscope, pretreatment with β-asarone or nimodipine dramatically increase the number of cells and improved the cellular morphology. Autophagosomes were found in OGD/R-treated PC12 cells as well as in drug plus OGD/R-treated PC12 cells.
  
  
  CONCLUSIONS
  β-Asarone protects PC12 cells against OGD/R-induced injury partly due to attenuating Beclin-1-dependent autophagy caused by decreasing [Ca(2+)](i) and increasing MMP.
authors.affiliation: The First Affiliated Hospital of Guangzhou University of Chinese Medicine, China.
authors.forename: Zhen-tao
authors.initials: ZT
authors.lastname: Mo
authors.forename: Yong-qi
authors.initials: YQ
authors.lastname: Fang
authors.forename: Yu-ping
authors.initials: YP
authors.lastname: He
authors.forename: Sheng
authors.initials: S
authors.lastname: Zhang
mesh_terms: D029023:Acorus / Q000737:chemistry*; D000081248:Allylbenzene Derivatives; D000818:Animals; D000840:Anisoles / Q000302:isolation & purification / Q000494:pharmacology*; D051017:Apoptosis Regulatory Proteins / Q000378:metabolism*; D001343:Autophagy / Q000187:drug effects*; D000071186:Beclin-1; D002545:Brain Ischemia / Q000188:drug therapy* / Q000378:metabolism; D002118:Calcium / Q000378:metabolism; D002121:Calcium Channel Blockers / Q000494:pharmacology; D002470:Cell Survival / Q000187:drug effects; D005947:Glucose / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D018696:Neuroprotective Agents / Q000302:isolation & purification / Q000494:pharmacology*; D009553:Nimodipine / Q000494:pharmacology; D010100:Oxygen / Q000378:metabolism; D016716:PC12 Cells; D051381:Rats; D015427:Reperfusion Injury / Q000188:drug therapy* / Q000378:metabolism
chemical_list: D000081248:Allylbenzene Derivatives; D000840:Anisoles; D051017:Apoptosis Regulatory Proteins; D000071186:Beclin-1; C512829:Becn1 protein, rat; D002121:Calcium Channel Blockers; D018696:Neuroprotective Agents; C012195:asarone; D009553:Nimodipine; D005947:Glucose; D010100:Oxygen; D002118:Calcium