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Poster P4:: Wednesday Posters significantly reduced by sulindac sulfide and ibuprofen but not naproxen and sulindac sulfone. Conclusions: We suggest that Fbxo11 may be a common and selective downstream target of A␤42 lowering NSAIDs. We are currently characterizing these candidate proteins for their functional significance for Alzheimer’s disease pathogenesis. P4-273

PHARMACOLOGICAL STUDIES OF TRADITIONAL CHINESE MEDICINE TO TREAT ALZHEIMER DISEASE

Lin Li, Lan Zhang, Ling Zhao, Wen Wang, Xuan-wu Hospital of Captical University of Medical Sciences, Beijing, China. Contact e-mail: [email protected] Background: Shen-wu Capsule (SWC) is a new compound containing 6 kinds of traditional Chinese herbs. TSG is a major effective component of SWC. Objective: To investigate the pharmacological effects of SWC and TSG on multiple kinds of AD-like animal models. Methods: The drugs were intragastrically administered to the animal models for 1 or 2 months. Morris water maze test was used to detect learning/memory ability, microarray and RT-PCR to measure gene expression, Western blotting and immuno-histochemistry to determine related proteins. Results: (1) In APP transgenic mice, SWC and TSG improved learning/memory ability, and decreased A␤ content and ␤-secretase expression; (2) in A␤ brain injection model, SWC and TSG inhibited microglial activation, and decreased IL-1␤ and tumor necrosis factor ␣ (TNF␣) content; (3) in ibotanic acid-induced basal forebrain cholinergic damaged rat model, SWC and TSG improved learning/memory ability, increased the ratio of cholinacetyl-transferase (ChAT)/cholinesterase (AchE), or enhanced M-receptor density; (4) in naturally aged rats of 24 months old, SWC improved learning/memory ability, decreased cholinergic cell death, and increased expression of nerve growth factor (NGF) and its receptor trkA in hippocampus; (5) in Dgalactose-induced brain aging mouse model, SWC and TSG improved learning/memory ability, inhibited lipid peroxidation, increased activities of antioxidant enzymes, and enhanced NGF and trkA expression in hippocampus; (6) in dementia rat model induced by mitochondria complex IV inhibitor azide perfusion, SWC improved learning/memory ability, increased the ratio of ChAT/AchE, decreased A␤ and hyperphosphorylated tau content, increased expression of protein phosphatase 2A (PP2A), brain derived neurotrophic factor (BDNF) and its receptor trkB; (7) in diabeticischemic model rats, SWC improved learning/memory ability, enhanced long term potentiation (LTP) in hippocampus, decreased cholinergic cell death, enhanced the expression of neurotrophin-3 (NT-3) and trkC, increased Bcl-2 and Akt protein expression, and decreased the content of Bax, caspase-3 and phosphated cAMP response element binding protein (P-CREB); (8) in hypercholesterolemia-induced dementia rat model, TSG improved learning/memory ability, decreased the content of A␤ and serum cholesterol. Conclusion: SWC and TSG act on multiple targets in the complicated pathogeneses of AD and thus may have strong potential to aid the treatment of AD. P4-274

COPPER-DEPENDENT UPTAKE OF CLIOQUINOL IN NEURONAL CELLS IS PROMOTED BY ␤AMYLOID

Shayne A. Bellingham1,2, Carlos Opazo1,3, James Camakaris2, Colin L. Masters1, Robert Cherny1, Ashley I. Bush1, 1Oxidation Disorders Laboratory, The Mental Health Research Institute and Department of Pathology, The University of Melbourne, Parkville, Victoria, Australia; 2Department of Genetics, The University of Melbourne, Parkville, Victoria, Australia; 3Laboratory of Neurobiometals, Department of Physiology, University of Concepcio´n, Box 160-C, Chile. Contact e-mail: [email protected] Background: Transition biometals, such as copper (Cu), zinc (Zn) and iron (Fe), play a central role in several neurodegenerative disorders including Alzheimer’s disease (AD). Perturbed Cu, Zn, and Fe homeostasis in AD brain tissue is believed to influence aggregation and toxicity of the

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␤-amyloid peptide (A␤). A␤ can bind both Cu and Zn with high affinity, resulting in aggregation of the toxic redox-active A␤ species. Thus, metal sequestration has been proposed as a therapeutic strategy for AD. To this extent, Clioquinol (CQ) has been shown to be effective in targeting metal interactions with A␤. CQ rapidly dissolves aggregates of A␤ in vitro, inhibits amyloid pathology in transgenic mice and rescues toxicity of A␤ in neuronal cell culture. Clinical trials of CQ therapy in AD patients report a slowing of cognitive decline accompanied by a lowering of plasma A␤1-42 levels. Since CQ is a lipid soluble compound that penetrates the blood-brain barrier, then potentially CQ-metal complexes may also penetrate into neurons in the brain. Objective(s): To investigate the neuronal cellular uptake of CQ in the presence of “free” or complexed biometals. Methods: Utilizing a cell culture system, we incubated mouse neuroblastoma cells (N2A) with radioiodinated CQ ([125I]CQ) alone or [125I]CQ in the presence of “free” Cu, Zn, and Fe or Cu-A␤ and Zn-A␤ complexes. The neuronal cellular uptake of [125I]CQ was then determined by a gamma counter. Results: We observed an increased uptake of [125I]CQ in the presence of both “free” Zn and Cu, but not Fe. The addition of 10 ␮M CuCl2 or 10 ␮M ZnCl2 resulted in a 2-3 fold increase in cellular [125I]CQ. Furthermore, when Cu or Zn was complexed to A␤1-42 we observed enhanced [125I]CQ uptake resulting in a 6-fold increase in the presence of 10 ␮M aggregated Cu-A␤1-42, but not 10 ␮M Zn-A␤1-42 or 10 ␮M soluble A␤1-42. Conclusions: These data support a novel mechanism for the therapeutic benefits of CQ, whereby CQ may not only bind bioavailable “free” copper but also remove Cu from aggregated A␤ plaques potentially restoring the imbalance in neuronal copper homeostasis. P4-275

UNCARIA RHYNCHOPHYLLA, A CHINESE MEDICINAL HERB, HAS POTENT ANTIAGGREGATION EFFECTS ON ALZHEIMER’S AMYLOID ␤ PROTEINS

Hironori Fujiwara1, Koh Iwasaki1, Katsutoshi Furukawa1, Takashi Seki1, Yukitsuka Kudo2, Yasushi Ohizumi3, Hiroyuki Arai1, 1 Department of Geriatric and Complimentary Medicine, Center for Asian Traditional Medicine Research, Tohoku University School of Medicine, Sendai, Japan; 2Tohoku University Biomedical Engineering Research Organization, Sendai, Japan; 3Department of Pharmaceutical Molecular Biology, Graduate School of Pharmaceutical Sciences, Tohoku University, Sendai, Japan. Contact e-mail: fujiwara@geriat. med.tohoku.ac.jp Background: Deposition of amyloid ␤ protein (A␤) is a consistent pathological hallmark of Alzheimer’s disease (AD) brains and A␤ accumulation begins at a relatively early stage in AD patients. Therefore, inhibition of A␤ generation, prevention of A␤ fibril formation, or destabilization of preformed A␤ fibrils would be attractive therapeutic strategies for the treatment of AD. Objective(s): To find novel therapeutic agents useful for Alzheimer’s disease from medicinal herbs which were used in traditional Chinese medical formula, in the present study, we examined the effects of Uncaria rhynchophylla on the formation or destabilization of A␤ fibrils. Methods: In order to determine whether this herb has the inhibitory effect on A␤ fibril formation and the destabilizing activity on preformed A␤ fibrils, we performed further thioflavin T binding assay, atomic force microscope imaging and electrophoresis. Results: Our study demonstrated that several of these herbs have potent inhibitory effects on fibril formation of both A␤1-40 and A␤1-42 in concentration-dependent manners, and particularly Uncaria rhynchophylla inhibited A␤ aggregation most intensively by 10 ␮g/ml of extract more than 50%. Significant destabilization of preformed A␤1-40 and A␤1-42 fibrils was also induced by Uncaria rhynchophylla as well as some other herb extracts. Three-dimensional HPLC analysis indicated that the water extract of this herb contains several different chemical compounds including oxindole and indol alkaloids, which has been regarded as neuroprotective on literature. To visualize A␤ fibril breakdown by Uncaria rhynchophylla, the fibrils were observed by the atomic force microscope. After incubation of A␤1-42 with the water extract of Uncaria rhynchophylla, preformed A␤ fibrils were largely di-