objective of the current research work was to evaluate Cilengitide trifluoroacetate the neuroprotective effect of the ethanol Cilengitide trifluoroacetate extract of (S. flavone; apigenin; luteolin; 6-hydroxyluteolin and so forth [12-16]. The phytochemicals present inScutellariaspecies have been reported to show a range of neuroprotective effects. Wogonin inhibited inflammatory activation of microglia by reduced cytotoxicity towards cocultured PC-12 neurons supporting anin vitroneuroprotective role of this flavonoid. The efficacy of wogonin was further demonstrated in two experimental brain injury models. In the 4-vessel occlusion model of transient global ischemia Rabbit polyclonal to Plexin B1. wogonin decreased the death rate of hippocampal neurons the induction of iNOS and TNF-in hippocampus whereas in the kainate injection model this flavonoid markedly protected from excitotoxic brain injury. Similarly baicalein attenuated the NO production by suppressing iNOS induction in LPS-activated BV-2 mouse microglial cells besides reducing apoptotic cell death and NF-kB activation [17-19]. 2 Materials and Methods 2.1 Materials The plant material (roots) ofScutellaria baicalensiswas collected from a local region of Hangzhou and was authenticated by a well-known botanist. Minimum essential medium (MEM) horse serum and fetal calf serum were obtained from Gibco. Multiwell plates were bought from Falcon. Laminin poly-L-lysine L-glutamine Glu glucose NMDA polyethylenimine and cytosine arabinoside were purchased from Merck. [3H]MDL 105 519 and [3H]MK-801 were purchased from Amersham Biosciences Inc. and MOLEKULA Ltd. respectively. All other chemicals were of reagent grade. 2.2 Preparation of the Extract The roots of the plant were thoroughly washed with tap water shade dried and then chopped into small pieces. Ethanol (95%) was used for hot extraction which was carried out for 4 hours using a soxhlet extraction apparatus. The extract was then concentrated under reduced pressure in a rotary evaporator at 40°C and was then kept in a refrigerator at 4°C prior to use. 2.3 Primary Rat Cortical Neuronal Cultures Primary rat cortical neuronal cultures were obtained from Sprague-Dawley (SD) Cilengitide trifluoroacetate rat embryos at embryonic stage of 14-16 days (Experimental Animal Centre of Sichuan University Chengdu City Sichuan Province China). The rats used in the experiment weighed between 250 and 300?g. The cerebral cortices were dissected and mechanically dissociated into single cells by trituration through Pasteur pipettes. Cells were plated at a density of 6 × 105 cells per well Cilengitide trifluoroacetate on 24-well culture plates coated with Laminin and poly-L-lysine. Then the cell cultures were incubated at 37°C in a humidified atmosphere of 5% CO2 in an MEM containing medium supplemented with glucose (25?mM) fetal calf serum (5%) horse serum (5%) and glutamine (5?mM). After 14-16 days in the culture medium the cells were used for the experiment. 2.4 Induction of Neuronal Cell Excitotoxicity and Their Assessment Earle’s balanced salt solution (EBSS) was used to rinse Cilengitide trifluoroacetate the cultured neuronal cells before the excitotoxic injuries were induced by exposure to 350?≤ 0.05. 4 Results 4.1 Assessment of Neuronal Excitotoxicity After the exposure of cultured rat cortical neuronal cultures to 35?S. baicalensisas claimed in the traditional Chinese medicine where this plant has been used against various neurological disorders. Our results demonstrated that when the neuronal cell cultures were exposed for 20?min to Glu (350?Scutellaria baicalensis(S.B.). Lactate dehydrogenase (LDH) activities given off by the damaged neurons into..