title: Neuroprotective effects of black soybean anthocyanins via inactivation of ASK1-JNK/p38 pathways and mobilization of cellular sialic acids.
abstract: OBJECTIVE
  To investigate neuroprotective effects of three major anthocyanins (cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, and petunidin-3-O-glucoside) isolated from the black soybean (Glycine max L.) cv. Cheongja 3 seed coat against H(2)O(2)-induced cell death of human brain neuroblastoma SK-N-SH cells.
  
  
  METHODS
  Cell viability, reactive oxygen species (ROS) generation, production and expression of heme oxygenase (HO)-1 and inactivation of mitogen-activated protein (MAP) kinase cascades were determined by MTT assay, 2,7-dichlorofluorescein diacetate (DCF-DA) assay, reverse transcriptase polymerase chain reaction (RT-PCR), and western blotting, respectively.
  
  
  RESULTS
  Pretreatment with anthocyanins reduced the cytotoxicity of H(2)O(2) on SK-N-SH cells, dose-dependently reduced the intracellular ROS level and inactivated apoptosis signal-regulating kinase (ASK1, Thr845), p38, and c-Jun N-terminal kinase (JNK) proteins. The HO-1 and Neu1 mRNA levels were increased by H(2)O(2) (25 μM) and further elevated by the pretreatment with anthocyanins. Sialic acids added to the culture plates not only attenuated the cytotoxicity of H(2)O(2) (25 μM) but also reduced intracellular ROS level. These results suggest that Cheongja 3 black soybean seed coat anthocyanins have brain neuroprotective effects against oxidative stress (H(2)O(2)) by inhibiting the activation of ASK1-JNK/p38 pathways, scavenging ROS, stimulating the expression of HO-1 and, more interestingly, recruiting cellular free sialic acids through up-regulation of Neu1 sialidase gene expression.
  
  
  CONCLUSIONS
  This is the first report indicating potent health benefits of black soybean seed coat anthocyanins in neuroprotection by triggering mobilization of cellular free sialic acid and utilizing it as an additional biological antioxidant in brain neural cells.
authors.affiliation: Department of Biotechnology, The Catholic Institute for Advanced Biomaterials, The Catholic University of Korea, Bucheon, Gyeonggi-do 420-743, Republic of Korea.
authors.forename: Sung Min
authors.initials: SM
authors.lastname: Kim
authors.forename: Mi Ja
authors.initials: MJ
authors.lastname: Chung
authors.forename: Tae Joung
authors.initials: TJ
authors.lastname: Ha
authors.forename: Ha Na
authors.initials: HN
authors.lastname: Choi
authors.forename: Seong Jae
authors.initials: SJ
authors.lastname: Jang
authors.forename: Sung Oog
authors.initials: SO
authors.lastname: Kim
authors.forename: Myung Hoon
authors.initials: MH
authors.lastname: Chun
authors.forename: Su Il
authors.initials: SI
authors.lastname: Do
authors.forename: Young Kug
authors.initials: YK
authors.lastname: Choo
authors.forename: Yong Il
authors.initials: YI
authors.lastname: Park
mesh_terms: D000872:Anthocyanins / Q000302:isolation & purification / Q000494:pharmacology*; D000975:Antioxidants / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D005786:Gene Expression Regulation / Q000187:drug effects; D005960:Glucosides / Q000302:isolation & purification / Q000494:pharmacology*; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D048031:JNK Mitogen-Activated Protein Kinases / Q000378:metabolism; D048848:MAP Kinase Kinase Kinase 5 / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism; D018696:Neuroprotective Agents / Q000302:isolation & purification / Q000494:pharmacology*; D018384:Oxidative Stress / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism; D012794:Sialic Acids / Q000378:metabolism / Q000494:pharmacology; D013025:Glycine max / Q000737:chemistry*; D048051:p38 Mitogen-Activated Protein Kinases / Q000378:metabolism
chemical_list: D000872:Anthocyanins; D000975:Antioxidants; D005960:Glucosides; D018696:Neuroprotective Agents; D017382:Reactive Oxygen Species; D012794:Sialic Acids; C462279:cyanidin-3-O-beta-glucopyranoside; C494120:delphinidin 3-O-glucopyranoside; C455107:petunidin-3-glucoside; D006861:Hydrogen Peroxide; D048031:JNK Mitogen-Activated Protein Kinases; D048051:p38 Mitogen-Activated Protein Kinases; D048848:MAP Kinase Kinase Kinase 5; C482858:MAP3K5 protein, human