Tibetan Medicine The Extract Wang Lahti CE A�� <sub> 25-35 </ Sub>-mediated Rat Neurons Toxic Injury

Wangla is a traditional Tibetan medicine and is the dry tuber of Coeotgolssum viride (L.) Hartm. var. barctaetum (Wild.) Rihcter belonged to the Orcidaceae family, which has effects such as complement of vigor and blood, helping produce saliva and slake thirst, smoothing the nerves, increasing intelligence and other. Pharmacological studies have shown that Coeotgolssum viride (L.) Hartm. var. barctaetum (Wild.) Rihcter extract (CE) could effectively inporving learning and merory in dementia mice and adbacute aging mice.In this study, the method of primary cultures of neonatal rat prefrontal cortex (PFC) neurons in vitro was used to study the effect of CE on the neurotoxic injury induced by β-amyloid peptide (Aβ), immunohistochemical β-tubulin III and DAPI method was used to identify the neuron purity of PFC, MTT and TUNEL was applied to observed the neuronal viability and death respectively, immunohistochemistry was used to detect the structure and expression of β-tubulin III. This study provides new data for studying the protective effects and mechanism and supports the benefit of CE on neruonal protection. The main conclusions are below: 1. through immunohistochemical β-tubulin III and DAPI on the neonatal rat PFC neurons cultured for5days, the positive staining for (3-tubulin III was green cells, showed better cell morphology, positive staining for DAPI was blue, experimental results showed that the neuron purity of PFC achieved95%.2. observing the neuronal viability and death through MTT and TUNEL found that treat with20μmol/L Aβ25-35for24hours in cultured neonatal rat PFC neurons had obvious neuronal toxicity, including the decline of cell viability and increase of cell apoptosis rate.3. MTT and TUNEL also found that pretreatment with CE before Aβ25-35treatment could suppress the neurotoxicity by Aβ25-35in cultured neonatal rat PFC neurons, different concentrations of CE (0.1mg/mL,1mg/mL,10mg/mL) have dose dependent protective effect on the injury of neuronal toxicity mediated by Aβ25-35.4. detecting neuron-specific marker β-tubulin III found obvious decrease of β-tubulin III staining in neonatal rat PFC neurons treated willi Aβ25-35and CE could partially restore β-tubulin III staining.