Swainsonine Induces The Apoptosis Of SD Rodent Nerve Cells

Swainsonine (SW), a polyhydroxy-substituted indole alkaloids, the chemical name is1,2,8-trihydroxyocta-hydroindolizidine.Swainsonine is the main toxic ingredient oflocoweed that can cause the animal locoweed poisoning， typical neurological symptomsand pathological changes are giving to the livestock,the main symptoms of chronicpoisoning can cause the head level tremor. In China locoweed mainly of the genus Oxytropisand Astragalus genus of poisonous plants, mainly distributed in the vast grasslands ofwestern China, and the hazard area has more than10%of the total area of grassland.At home and abroad,there is not enough research on the cellular and molecular level oftoxicity and the mechanism of toxicity on swainsonine.swainsonine neurotoxicity is not veryclear understanding,making the control technology of locoweed poisoning disease is slowlydeveloped. This test is intended on the basis of previous research, Establishment of newbornSD rat cerebral cortical neurons in vitro model to detect swainsonine toxicity and apoptosis ofnerve cells, for further research on the molecular mechanisms of neurotoxicity and locoweedpoisoning mechanism of toxicity.1The primary culture methods to creat the moden of vitro newborn rat cerebralcortical neurons and identification of its activity. through improving the existing nervecells and promoting neuronal growth and development,using the serum-free culture mediumand cytarabine to inhibite the growth of other cells. The results is:MTT assay activity of thestrongest is the5thd, Nissl’s staining detection, the purity of the neurons increased from79%to92%. So we have established the relative stability of culture the nerve cells in vitro.the cellculture model can be used for further research on the neural mechanisms of toxicity ofswainsonine.2The morphology reseach of SD rat cerebral cortex nerve cell injuried byswainsonine. Having test of different concentrations of swainsonine on the survival of nervecells at different time, according to toxicology-related research methods to determine24h,IC50(0.8511mg/mL), filter out the three swainsonine concentrations (0.5mg/mL,1mg/mL,2mg/mL) for12h as the next step to study the toxic effects of swainsonine model. We havefound the morphological study of the three concentration dose group compared with the control group in vitro culture of cerebral cortical neurons cell have decreased with increasingconcentration of swainsonine, the processes also shorten and the cell density were decreased,the network formation decreased, the change is more obvious, This means there are obviousdose-effect of the toxic effects by swainsonie to nerve cells.3Swainsonine induced SD rat cerebral cortical neurons apoptosis. using nuclearstaining techniques, flow cytometry determination of apoptosis rate and cell cycle, we findthe toxic effects of swainsonine on nerve cells: with increasing concentration of swainsonine,the nerve cell nucleus narrow, crescent-shaped chromatin condensation, and in severeapoptotic bodies, nuclear fragmentation and significantly reduced the number of normal cells;the G0/G1of the cell decreased and S phase cells gradually increased; early apoptotic cellswere significantly increased (P<0.05), late apoptosis is also increasing, there is a significantdose-effect relationship. Prove that low doses of swainsonine induced apoptosis, whereashigh doses can directly kill cells.