The Effect Of Cultured Astrocyte On The Damaged Neurons Induced By Simulated Cerebral Ischemia And Reperfusion In Vitro And Influence Of Kangdai â… 

It has very important meaning to the prevention and cure of cerebrovascular disease to studythe characteristic, disciplinarian and mechanism of the damage induced by cerebral ischemiareperfusion. The damage induced by cerebral ischemia reperfusion was investigated solely fromneuron or neuroglia in the past, however they are the functional whole and should be studiedtogether. In recent years particularly ischemia preconditioning(IPC) has beening concerned as ainherent sheltered mechanism by the people. During brain ischemia preconditioning(BIP) neuronsshow cerebral ischemia tolerance and astrocytes play a significant protective role. Based on thisviews ,we investigated the relation between astrocytes and neurons after ischemia andreperfusion ,and then studied the effect of Kangdai I through experiments.We researched the effectof cultured astrocyte conditioned medium on the damaged neurons induced by simulated cerebralischemia and reperfusion in Vitro and influence of Chinese herbal Kangdai I with the technique ofseparation and purification to cultured neurons and astrocytes. The aim is to probe into themechanism and regulation of cerebral ischemia and reperfusion injury and BIP ,so as to providemore scientific and reliable experimental evidences to the treatment of Chinese herbs forcerebrovascular diseases and the research & development of new medicine. In the experiment ,the neurons and astrocytes cultured cerebral cortex of rats wereresearched, based on simulated cerebral ischemia and reperfusion in vitro. We investigated theavtivity of cultured neurons and the secretory function of cultured astrocyte in the damage inducedby simulated cerebral ischemia and reperfusion in vitro, then studied the effect of astrocyteconditioned medium(ACM) on injured neurons and the influence of Kangdai I with biochemicalmethod and immunocytochemical technique .The main results and conclusions are as follows:1. Cultured neurons were damaged by simulated cerebral ischemia and reperfusion in vitro: Results: (1)The activity of the damaged neurons was decreased, the survival rate was descent. (2)The transudation rate of LDH in cultured medium increased obviously. (3)The mortality was increased significantly. (4) The strong positive cell count of NOS expression significantly was increased 4h afterischemia (Is-4h)and 3h after reperfusion (Rp-3h). Conclusion:The reason of neurons injury may be : (1) The injury of mitochondria results in disturbance of energetic metabolism. (2) The damage of cell membrane structure induces membrane infiltrative change to cause devastating continuous biochemical reaction.-4- 培养大鼠星形胶质细胞对拟脑缺血再灌注损伤神经元的作用和抗呆Ⅰ号的影响 (3) The increase of NO and its derived toxic free radicals cause the toxic harm on neurons.2. Cultured astrocytes were damaged by simulated cerebral ischemia and reperfusion in vitro: Results: (1)The activity of the damaged astrocyte was decreased, the survival rate was descent. (2)The ability of secreting total protein decreased. Conclusion: Cultured astrocytes was damaged in the model simulated cerebral ischemia andreperfusion in vitro.3. There were certain regularities in the change of damaged neurons and astrocytes induced bysimulated cerebral ischemia and reperfusion in vitro, it could be divided into: (1) Cell injury period: from the begin of ischemia to Rp-3h. (2) Functional compensation period: from the end of Rp-3h to Rp-18h. (3) Lower functional period: from the end of Rp-18h to Rp-36h. (4) Functional restoration period: from the end of Rp-36h to Rp-72h.4. The ability of astrocytes secretory was increased in the damage induced by simulated cerebralischemia and reperfusion in vitro, its high peak of expression was respectively as follows: (1) BDNF:The end of Rp-36h. (2) GDNF: from the end of Rp-3h to Rp-18. (3) bFGF:The end of Rp-3h. (4) HSP70:The end of Rp-48h. (5) IL-6: from the end of Rp-18h...