Abstract Title

ROLE OF ALCOHOL IN PRIMARY HUMAN ASTROCYTES

Presenter Name

Richa Pandey

Presentation Type

Poster

Purpose (a):

The aim of this study was to investigate alcohol-mediated activation of human astrocytes and subsquential alterations in their inflammatory functions.

Methods (b):

Primary human astrocytes were incubated with or without alcohol at doses of 25, 50, 100 mM for 3, 5, and 7 days. Cells and culture supernatants were collected. Astrocyte morphology was examined by immunocytochemical staining for glial fibrillary acidic protein (GFAP). Expressions of TIMP-1 and pro-inflammatory cytokines including CXCL8 and CCL2 were measured by ELISA. Cell metabolic activity, proliferation, and apoptosis were analyzed by MTT assay, BrdU cell proliferation assay, and Cell Death Detection ELISAplus assay, respectively.

Results (c):

Alcohol exposure altered the morphology of astrocytes to a reactive phenotype as determined by GFAP immunostaining. Alcohol significantly upregulated TIMP-1 levels in dose-dependent manner (P< 0.05), with a peak at day 5 post-treatment. Moreover, alcohol treatment significantly upregulated CCL2 (P< 0.05) while CXCL8 was significantly downregulated in a dose as well as time-dependent manner (P< 0.05). In addition, alcohol exposure significantly decreased astrocytes viability (P< 0.05) and proliferation as measured by MTT and BrdU incorporation assay respectively, and significantly increased apoptosis (P< 0.05) in parallel experiments.

Conclusions (d):

In summary, our results suggest that alcohol may alter astrocyte inflammatory mediators and/or regulate astrocyte functions.

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ROLE OF ALCOHOL IN PRIMARY HUMAN ASTROCYTES