| Depression is a common spirit aplastic disease, it has affected nearly1/5of the population all over the word. So, the pathogenesis of depression and treatment research have got the more and more attention of the scientific research workers. At present, the antidepressant drugs are mainly synthetic drugs, but they have a large number of drawbacks, such as long time action, drug resistance, severe side effects, and so on. So the development of safety, high efficiency, low natural antidepressant drugs become this hot spot of this area research. Pigeonpea is a major beans grown in tropical and subtropical areas. The previous reports on pigeonpea focused on nutrition compositions, and only in recent years involve medical compositions. This study is to investigate antidepressant activity effect of Cajanus cajan, on this basis to study its antidepressant effect components and the possible mechanisms, so as to provide the theory and experimental base to the development of new antidepressant drugs.Aim of the studyTo investigate Cajanus cajan neuroprotective effect against the toxicity induced with corticosterone in PC12cells, as well as the possible mechanisms.Materials and methods1. Use corticosterone-induced PC12cells as drug screening model in vitro.2. After alcohol extracts and its fractions from Cajanus cajan treatments, cell viability was determined by incubating with MTT assay and evaluated by measuring the amount of cytoplasmic lactate dehydrogenase(LDH) released into the medium.3. In this study ethyl acetate parts and ethanol parts were separated.4. The isolated compounds were screened by incubating with MTT assay.5. Through testing the LDH leaks out quantities and DNA fragments to observe the active ingredients effect to corticosterone-induced PC12cells apoptosis.6. The intraeellular [Ca2+] in PC12cells were tested in the study by fluorospectrophotometry. 7. Use Caspase-3kit to detect Caspases-3protease express amounts of corticosterone-induced PC12cells.Results1. PC12cells morphological result showed that cultured PC12cells were treated with100μmol/L corticosterone for48h resulted in obviously cell loss and neurites injury. Exposure of cells to corticosterone for48h, cells survival rate became to60.5%by MTT assay.2. corticosterone-induced PC12cells were counteracted by100-12.5mg/L alcohol extracts from Cajanus cajan and p<0.01has significant differences.3. Ethyl acetate parts in100-12.5mg/L and ethanol parts in200-12.5mg/L had significant protective activity in time and dose dependent relationship.4.12compounds were separate out from acetate parts and ethanol parts. Exposure of cells to corticosterone for48h, cells viability were60.1%, then treatment longistyline A (1.2,2.4,4.8μg/L) for24h, cell viability were67.95%,71.46%and72.68%respectively. Treatment CSA(0.6,1.2,2.4,μg/L) for24h, cell viability were82.36%,84.90%and93.48%respectively, p<0.01has significant differences.5. Through testing the LDH leaks out quantities and DNA fragments, the study observed the longistyline A and CSA could significantly reduce the LDH leaks out quantities and DNA fragments.6. The study results showed the intracellular [Ca2+] in PC12cells significantly increased after corticosterone treatment compared to the normal level(P<0.01). Treatment of longistyline A and CSA, after corticosterone induced PC12cells, Fura2-AM fluorescence intensity decreased significantly compared with the model level.7. Caspase-3test results showed that the model group Caspase-3OD value increased significantly compared with blank control group, statistically significant (P<0.01); After corticosterone induced treatment of longstyl A and CSA decreased the OD value.ConclutionsCajanus cajan and monomer longistyline A and CSA could generate a neuroprotective effect on corticosterone-induced neurotoxicity in PC12cells possible by decreasing the [Ca2+]i concentrations and Caspase-3activity. |
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