Free -radical generation by amyloid p protein and the protective effect by laminin against amyloid β-protein induced toxicity

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We investigated the inhibitory effect by laminin, which is one of the extracellular matrix components, against beta amyloid(Aβ) induced free radicals by electron paramagnetic resonance(EPR) using PC12 cells with two kinds of spin trap, 5,5-dimethyl-1-pyrroline-N-oxide ; DMPO, α-(4-pyridyl-1-oxide)-N-tert-butylnitrone ; POBN respectively. We investigated two kinds of experiment to clear the mechanism how laminin shows the protection against Aβ. At first, EPR measurement was performed to see whether or not laminin directly inhibit Aβ induced free radicals, Aβ was mixed with laminin and the solution was added to PC12 cells. The second, EPR measurement was also performed to see whether or not the cells which attached laminin get strengthened their defense function.The free radical production was reduced when Aβ was mixed with laminin and the cell viability also increased compared with Aβ only added to the cells, though there was no difference in generating free radicals between cells which were attached with laminin for 24hrs before Aβ addition and cells which did not attach laminin for 24hrs. Therefore we concluded that laminin has neuroprotective ability and its protective effect against Aβ induced free radical generation is due to possibly direct reaction with Aβ.