Tongluojiu'nao The Protective Effect Of Brain Injection Of Rat Hippocampal Neurons

In the modern medical study of encephalopathy,especially in the development of molecular genetic technology,an increasing recognition that the relationship between encephalopathy and brain micro-environment(whether cerebral ischemia or neurodegenerative diseases) is much closed."Toxic hurts brain collaterals" is the key point in its pathogenesis,and remarkable clinical curative effect has obtained with the treatment targeting on the functional characteristic of collateral. TongLuoJiuNao(TLJN) is a herb extract that mainly contains ginsenoside Rg1 and geniposide which are clinically used for treating ischemic damages in the brain, however,the mechanism of this protection is unclear yet.Futhermore,under the instruction of "collateral theory" of Chinese medicine and focus the target on the Brain Microvascular Endothelial Cells(BMEC).We further studied the regulatory influence of rat brain microvascular endothelial cells on the activity of cortical neurons.The research focus on the neuroprotective effects which by using the cell model of oxygen glucose deprivation and amyloid protein in hippocampal neurons,the objective is to observe TLJN injection and its different components whether can rescue the nerve injury of cerebral ischemia andβ-amyloid neurotoxicity.The following two parts are related experiments:The first part,to observe TLJN injection can or cannot rescue neurons by cerebral oxygen glucose deprivation through brain microvascular endothelial cells,and to explore TLJN injection's intervention effects on cerebral ischemic injury after stroke.The second part,to deeply study the neuroprotective effect of TLJN injection and its different components in cell model can or cannot rescue the neurotoxicity of amyloid protein.1,Identificated isolated primary cultured rat brain microvascular endothelial cells and hippocampal neurons by using immunocytochemical.By the method of six hours of oxygen glucose deprivation to model the cerebral ischemia-reperfusion, and using LDH methods to observe the neuroprotective effect in oxygen glucose deprivation injury by compared with the TLJN injection and model group. 2,Collected conditioned medium of brain microvascular endothelial cells during the different periods of oxygen deprivation after TLJN injection treatment,and observed the indirect effects of drugs on the neuronal survival.We find that the normal conditions and reoxygenation group can markedly improved cell survival.3,Observed the indirect effects of drugs on the synapses growth of neurons in condition medium.We find that TLJN injection can promote synaptic extension in different levels of conditioned medium.It prompted that the drug can effect on brain microvascular endothelial cells and then promote them to secret some neurotrophic substances to promote synaptic growth.4,Online predict the TLJN injection and its different compounds,suggesting that they has neuroprotective effect.Adopted amyloid protein in neuronal cell model to model the Alzheimer's disease.The results showed that TLJN injection can protect the survival and can prevent the synaptic damage by using cell survival statistics and immunohistochemical.Geniposide's protective effect is more obvious while the Ginsenoside can obviously present the synaptic damage.In this study,we investigated whether TLJN can protect neurons against damages by ischemia in brain vasculature,and the effects was evaluated with cell viability and neurite outgrowth before or after OGD.We found that TLJN could protected both cultured primary rat hippocampal neurons and BMECs from cell death under both normal and oxygen/glucose-deprivation(OGD) conditions.Moreover,under the same conditions,BMECs-conditioned media pretreated by TNJN could also promote neuron viability and neurite outgrowth,indicating that TLJN stimulated BMECs to secret some neuroprotective/ neurotrophic factors.These findings suggest that TLJN has a marked neuroprotective and neurotrophic roles by either direct or indirect operation,and provide insight into the mechanism of clinical efficacy of this drug against stroke and AD.