title: Vitamin D2 suppresses amyloid-β 25-35 induced microglial activation in BV2 cells by blocking the NF-κB inflammatory signaling pathway.
abstract: OBJECTIVE
  Present emerging world is emphasizing the implication of vitamin D deficiency associated with development of inflammation and neurodegenerative disorder like Alzheimer's disease (AD). The chief neuropathological hallmark of AD is aggregation of amyloid-beta (Aβ) peptides surrounding microglial cells in human brain. Microglial activation plays a key role in inflammatory response and neuronal injury. Naturally abundant vitamin D2 (VD2) exhibiting anti-inflammatory activities are yet to explore more. This study has investigated the inhibitory effect of VD2 on inflammatory activities of BV2 microglial cells.
  
  
  METHODS
  Cellular compatibility of VD2 and Aβ25-35 protein in treated BV2 microglial cells were measured by CCK-8 assay. Induction of iNOS, COX-2 and NF-κB signaling cascade were measured by western blotting, whereas pro-inflammatory cytokines were measured by ELISA. In addition, generation of ROS was detected by fluorescence intensity.
  
  
  RESULTS
  Morphological observations showed that Aβ25-35 induced BV2 cells stimulation noticeably got reduced in VD2 pre-treated group at 24h time period. Anti-inflammatory activities of VD2 was observed demonstrating the inhibition of up-regulated iNOS and COX-2 protein expression further confirmed by attenuating the activated microglia released pro-inflammatory cytokines IL-1β, IL-6, TNF- α and ROS, while blocking the phosphorylation of NF-κB p65 in nucleus by preventing IκB-α degradation and phosphorylation in cytosol.
  
  
  CONCLUSIONS
  The present study revealed that VD2 blocked the phosphorylation of NF-κB inflammatory signaling pathway in Aβ25-35 induced activated BV2 microglial cells by suppressing ROS generation and inflammatory cytokines. Our finding suggests that vitamin D2 has therapeutic potential against inflammation and Alzheimer's disease.
authors.affiliation: Research Institute of Life Science and College of Veterinary Medicine (BK21 plus project), Gyeongsang National University, Gazwa, Jinju 660-701, Republic of Korea.
authors.forename: Suchismita
authors.initials: S
authors.lastname: Raha
authors.affiliation: Research Institute of Life Science and College of Veterinary Medicine (BK21 plus project), Gyeongsang National University, Gazwa, Jinju 660-701, Republic of Korea.
authors.forename: Ho Jeong
authors.initials: HJ
authors.lastname: Lee
authors.affiliation: Research Institute of Life Science and College of Veterinary Medicine (BK21 plus project), Gyeongsang National University, Gazwa, Jinju 660-701, Republic of Korea.
authors.forename: Silvia
authors.initials: S
authors.lastname: Yumnam
authors.affiliation: Research Institute of Life Science and College of Veterinary Medicine (BK21 plus project), Gyeongsang National University, Gazwa, Jinju 660-701, Republic of Korea.
authors.forename: Gyeong Eun
authors.initials: GE
authors.lastname: Hong
authors.affiliation: Research Institute of Life Science and College of Veterinary Medicine (BK21 plus project), Gyeongsang National University, Gazwa, Jinju 660-701, Republic of Korea.
authors.forename: Venu
authors.initials: V
authors.lastname: Venkatarame Gowda Saralamma
authors.affiliation: Department of Applied Chemistry, Institute of Agriculture and Life Science, Gyeongsang National University, Jinju 660-701, Republic of Korea.
authors.forename: Yeong Lae
authors.initials: YL
authors.lastname: Ha
authors.affiliation: HK Biotech, Co. Ltd., Jinju 660-844, Republic of Korea.
authors.forename: Jeong Ok
authors.initials: JO
authors.lastname: Kim
authors.affiliation: HK Biotech, Co. Ltd., Jinju 660-844, Republic of Korea.
authors.forename: Young Suk
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Gyeongnam Department of Environment Toxicology and Chemistry, Toxicology Screening Research Center, Korea Institute of Toxicology, Jinju 666-844, Republic of Korea.
authors.forename: Jeong Doo
authors.initials: JD
authors.lastname: Heo
authors.affiliation: Gyeongnam Department of Environment Toxicology and Chemistry, Toxicology Screening Research Center, Korea Institute of Toxicology, Jinju 666-844, Republic of Korea.
authors.forename: Sang Joon
authors.initials: SJ
authors.lastname: Lee
authors.affiliation: Department of Nursing Science, International University of Korea, Jinju 660-759, Republic of Korea.
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Kim
authors.affiliation: Research Institute of Life Science and College of Veterinary Medicine (BK21 plus project), Gyeongsang National University, Gazwa, Jinju 660-701, Republic of Korea. Electronic address: gonskim@gnu.ac.kr.
authors.forename: Gon Sup
authors.initials: GS
authors.lastname: Kim
mesh_terms: D016229:Amyloid beta-Peptides / Q000378:metabolism*; D000818:Animals; D002461:Cell Line, Transformed; D004872:Ergocalciferols / Q000494:pharmacology*; D006801:Humans; D051379:Mice; D017628:Microglia / Q000473:pathology*; D016328:NF-kappa B / Q000378:metabolism*; D019636:Neurodegenerative Diseases / Q000378:metabolism / Q000473:pathology; D010446:Peptide Fragments / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D015398:Signal Transduction
chemical_list: D016229:Amyloid beta-Peptides; D004872:Ergocalciferols; D016328:NF-kappa B; D010446:Peptide Fragments; D017382:Reactive Oxygen Species; C074636:amyloid beta-protein (25-35)
keywords: Alzheimer; Amyloid-β 25–35; Amyloid-β 25–35 (PubChem CID: 10843733); BV2 microglial cells; Ergocalciferol (PubChem CID: 5280793); H2DCFDA (PubChem CID: 77718); Inflammation; Nuclear factor-kappaB; Vitamin D(2)