title: At low doses ethanol maintains blood-brain barrier (BBB) integrity after hypoxia and reoxygenation: a brain slice study.
abstract: Post-ischemia ethanol (EtOH) treatments have been shown to exhibit neuroprotective effects in stroke. However, the mechanisms underlying these effects and those on blood-brain barrier (BBB) integrity have yet to be elucidated. In the present study, we determined whether administering differing concentrations of EtOH alter the expressions of BBB integral proteins, including aquaporins-4 and -9 (AQP-4, AQP-9), matrix metallopeptidases-2 and -9 (MMP-2, MMP-9), zonula occludens-1 (ZO-1), and basal lamina (laminin). We employed an organotypic brain slice culture model that utilizes oxygen-glucose deprivation followed by reoxygenation (OGD/R). Brain slices were obtained from 10-day-old Sprague-Dawley rats and divided into the following five groups (n = 8 subjects per group): (1) control, (2) hypoxia (OGD/R), no EtOH, (3) OGD/R and 10 mM EtOH, (4) OGD/R and 30 mM EtOH, and (5) OGD/R and 90 mM EtOH. To assess BBB integrity, levels of AQPs, MMPs, ZO-1, and laminin were determined by Western blot. Compared to control, OGD/R without EtOH significantly increased AQP-4, AQP-9, MMP-2, and MMP-9 levels, while decreasing ZO-1 and laminin levels. All EtOH concentration treatments (groups 3 through 5) significantly reduced the expressions of AQP-4, AQP-9, MMP-2, and MMP-9, compared to the OGD/R, non-alcohol treated slices. Furthermore, compared to the OGD/R without EtOH group, the 30 mM EtOH treatment significantly increased ZO-1 and laminin levels. In contrast, the 90 mM EtOH level neither enhanced the reduction in AQP and MMP levels nor increased ZO-1 or basal lamina expressions observed in the 30 mM treatment. In conclusion, at an optimal dose of 30 mM, EtOH improves the expressions of MMP-2, MMP-9, AQP-4, AQP-9, ZO-1, and basal laminin, previously altered by OGD/R. These effects may indicate a beneficial effect of EtOH on BBB integrity after stroke.
authors.affiliation: Wayne State University School of Medicine, Detroit, MI, USA.
authors.forename: Changya
authors.initials: C
authors.lastname: Peng
authors.forename: William A
authors.initials: WA
authors.lastname: Li
authors.forename: Paul
authors.initials: P
authors.lastname: Fu
authors.forename: Tia
authors.initials: T
authors.lastname: Chakraborty
authors.forename: Mohammed
authors.initials: M
authors.lastname: Hussain
authors.forename: Murali
authors.initials: M
authors.lastname: Guthikonda
authors.forename: Jose A
authors.initials: JA
authors.lastname: Rafols
authors.forename: Yuchuan
authors.initials: Y
authors.lastname: Ding
mesh_terms: D000818:Animals; D051401:Aquaporin 4 / Q000378:metabolism; D020346:Aquaporins / Q000378:metabolism; D001812:Blood-Brain Barrier / Q000187:drug effects*; D001921:Brain / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D000431:Ethanol / Q000494:pharmacology* / Q000627:therapeutic use; D005947:Glucose / Q000172:deficiency; D000860:Hypoxia / Q000188:drug therapy*; D007797:Laminin / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D018696:Neuroprotective Agents / Q000627:therapeutic use; D010100:Oxygen / Q000627:therapeutic use; D051381:Rats; D020521:Stroke / Q000188:drug therapy*; D062826:Zonula Occludens-1 Protein / Q000378:metabolism
chemical_list: C491637:Aqp9 protein, rat; D051401:Aquaporin 4; D020346:Aquaporins; D007797:Laminin; D018696:Neuroprotective Agents; D062826:Zonula Occludens-1 Protein; D000431:Ethanol; D020778:Matrix Metalloproteinase 2; D020780:Matrix Metalloproteinase 9; D005947:Glucose; D010100:Oxygen