22. European Stroke Conference 476 Experimental studies Effects of exogenous lithium on hypoxic endothelial cells C. Rodemer1, M.G. Hennerici2, S. Meairs3 UMM, Mannheim, GERMANY1, UMM, Mannheim, GERMANY2, UMM, Mannheim, GERMA-NY3 Background: Several reports have documented neuroprotective effects of lithium. These include re-duction of brain damage in animal models of neurodegenerative diseases and reduction of neurolog-ic deficits when administered during brain ischemia. Here we examined the effects of Lithium Chlo-ride (LiCl) on mouse brain endothelial cells (bEND.3) cells during hypoxia. Methods: bEND.3 cells were placed for 20h in a hypoxia chamber (Billups-Rothenberg) with a 95%N2/5%CO2 atmosphere in OptiMEM (Invitrogen) without fetal calf serum and glucose and with 10mM LiCl during hypoxia. Lucifer yellow (LY) flow-through experiments were performed using transwell inserts (Corning) and cell growth was determined (CCK-8, Dojindo). Expression of proteins was analyzed by Western blotting. Results: Lithium did not have a significant effect on proliferation of untreated cells (ctrl:0.68 OD450 +/-0.15; ctrl+LiCl:0.7+/-0.23). After 20h of hypoxia cell proliferation was reduced (0.53+/-0.11) but not totally recovered with LiCl (0.62+/- 0.23). LY flow through (cm/min) was reduced after hy-poxia (ctrl: 1.14+/-0.4; hypoxia: 0.86+/-0.14) but not restored with LiCl (0.87+/-0.17). Expression of pERK1/2 was increased in hypoxic cells (ctrl: 100%; hypoxia: 143%+/-26%) but decreased after addition of LiCl (128%+/-32%). Expression of peNOS was slightly reduced after hypoxia and mod-erately increased with LiCl (hypoxia: 87%+/-10;hypoxia+LiCl: 93%+/-16%). Catenin expression did not significantly change after hypoxia but it was lowered with LiCl (hypoxia: 101%+/-18,hypox-ia+ LiCl: 87%+/-34). Actin expression remained unchanged. Conclusion: Although LiCl was not able to fully recover endothelial cell proliferation after hypoxia, proliferation was partly enhanced by LiCl. Surprisingly, LY flow was not affected by LiCl. This may be because LiCl does not significantly affect expression of proteins like catenin or peNOS. In con-clusion exogenous LiCl could not regulate tightness of bEND.3 endothelial cell junctions. 540 © 2013 S. Karger AG, Basel Scientific Programme 477Experimental studies Neuroprotective and Neurogenesis effects of curcumin in the adult rat following transient global ischemia F.A. Attari1, H.A Aligholi2, G.H. Hassanzadeh3, A.L. Gorji4 Shefa Neuroscience Research Center, Khatamolanbia Hospital, Department of Neuroscience, School of Advanced Medical Technology, Tehran University, Tehran, IRAN1, Shefa Neurosci-ence Research Center, Khatamolanbia Hospital, Department of Neuroscience, School of Advanced Medical Technology, Tehran University, Tehran, IRAN2, Department of Anatomy, School of Med-icine, Tehran University of Medical Sciences, Iran, Tehran, IRAN3, Shefa Neuroscience Research Center, Khatamolanbia Hospital, Tehran, IRAN4 Background: Ischemic brain injuries still rank high througout the world. The recent findings have demonstrated the adult brain possesses neurogenesis potentiality. curcumin can reduce oxidative and stimulate neurogenesis in the brain. The aim of present study is evaluation of neuroprotective and neurogenesis effects of curcumin in the adult rat following transient global ischemia. Methods: Fourty-eight male wistar rats were randomly assigned to eight groups contain 6 rats. Con-trol group, sham groups (animals only with global ischemia), treatment groups (animals with 100 and 300 mg/kg curcumin administration) and the vehicle group. Sacrification was performed after 3 or 4 weeks. Animals underwent temporary ligation of bilateral common carotid artery. Histological features of neuronal injury were stained with nissl, TUNEL and Brdu. Results: the number of dark cells and apoptotic ones in Isch 3W, Isch 4W and vehicle groups in-creased notably compared with control rats in all regions. Treatment with 100 mg/kg curcumin had few effects on decrease of dark cells and apoptotic ones after 3 weeks. The number of dark neurons and apoptotic ones in C100 4W decreased significantly in CA3, CA2 and temporal cortex. Treat-ment with 300 mg/kg curcumin both after 3 and 4 weeks had remarkable effects on decline of them. The number of BrdU-labeled cells in Isch 3W, Isch 4W and vehicle groups increased considerably in DG and temporal cortex. In C100 3W generated new cells increased strikingly in PPV and SVZ and in C100 4W increased extensively in all of the areas. The generation of new neurons decreased dramatically in C300 3W and 4W groups in all of the regions. Conclusion Global ischemia in the period of 3 and 4 weeks had destructive effects on hippocampus and temporal cortex. Curcumin in lower doses was so effective for neurogenesis. Cell proliferation started in common (SGZ, SVZ) and uncommon (PPV) neurogenic sites 1-2 weeks after transient global ischemia and curcumin administration but cell migration started after 3-4. Ultimately, cur-cumin in higher doses was neuroprotective enough but its effects on the generation of new neurons was converse.
Karger_ESC London_2013
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