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• W2104177211 abstract "A-beta deposits powerfully stimulate inflammation in AD brain. Agents that inhibit the buildup of these deposits, or assist in their phagocytosis should be effective therapeutic agents. We developed simple in vitro screening assays to identify such agents as a preliminary to in vivo testing. Inhibition of beta-amyloid aggregation. For details see J-P Guo et al., JAD 19; 2010. Test agents were mixed with a fluorophore labeled beta-amyloid 42 probe for 30 minutes at 37C. The mixture was then applied in the dark to plaque containing AD brain sections for one hour at room temperature. Binding of the A-beta fluorophore to A-beta deposits was observed microscopically. Inhibition of test agents was assessed quantitatively. Enhancement of phagocytosis. Uptake of FluoSphere beads (diameter 0.02um) by THP1 cells was assessed. Test agents and 1ul of 0.025% microsphere beads were added into wells containing 2X10 4 THP1 cells in 200ul of HBSS. The mixture was incubated in the dark for one hour at 37C. After washing with HBSS, the cells were smeared on glass slides, air-dried and fixed in methanol/acetone (1:1). The fluoro-phagocytosis signal was observed microscopically and enhancement of phagocytosis determined semi-quantitatively. For beta-amyloid phagocytosis, 1ul of 0.1mg/ml beta-amyloid-HiLyte488 probe and test agents were similarly added to wells and incubated for one hour at 37C. The cells were treated and observed as the described previously. Several thousand medicinal agents were tested for their ability to inhibit beta amyloid binding and several hundred for enhancing its phagocytosis by THP-1 cells. Some highlights were that 1mg/ml of 1,2,3,4,6-penta-O-galloyl-b-D-glucopyranose (PGG) completely blocked fluoro-beta-amyloid probe binding while 0.5ug/ml enhanced phagocytosis . 1mg/ml of resveratrol partially blocked fluoro-beta-amyloid probe binding while 50ug/ml enhanced phagocytosis. 5mg/ml of (-)-borneol almost completely blocked fluoro-beta-amyloid probe binding while 25ng/ml enhanced phagocytosis. 0.5mg/ml of epigallocatechin gallate (EGCG) almost completely blocked fluoro-beta-amyloid binding, but even at 50ug/ml it had no effect on phagocytosis. 0.5% ginger extract completely blocked fluoro-beta-amyloid binding but 2.5% completely blocked phagocytosis. PGG, resveratrol and borneol are attractive agents for in vivo testing because they can hinder beta-amyloid deposition while enhancing its phagocytosis by THP1 cells." @default.
• W2104177211 created "2016-06-24" @default.
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• W2104177211 date "2012-07-01" @default.
• W2104177211 modified "2023-10-16" @default.
• W2104177211 title "P1-330: Simple in vitro screening assays for agents that inhibit beta-amyloid deposition or promote beta-amyloid phagocytosis" @default.
• W2104177211 doi "https://doi.org/10.1016/j.jalz.2012.05.2016" @default.
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