Vitamin D2 suppresses amyloid-β 25-35 induced microglial activation in BV2 cells by blocking the NF-κB inflammatory signaling pathway

Abstract

Aims: 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.

Main 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.

Key findings: 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.

Significance: 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.

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).