中文摘要
本项目研究以全脑缺血或局灶脑缺血或缺糖缺氧（OGD）细胞模型，采用免疫沉淀，免疫印迹，细胞膜片钳技术等方法，研究了抑制性GABA信号对脑缺血诱导的兴奋性信号通路的调控机制。结果显示：（1）GABA(A)受体和GABA(B)受体激动剂联合用药，不但能抑制脑缺血诱导的GluR6介导的JNK通路的激活，而且还能抑制KA刺激引起JNK通路的激活；不但能抑制NMDA受体亚基NR2A酪氨酸磷酸化及增强nNOS的磷酸化，而且还可通过激活PI-3K/Akt通路抑制ASK1-JNK通路，从而达到保护神经元的作用。（2）GluR5的激动剂刺激突触前GluR5不但能抑制脑缺血诱导突触后GluR6介导的JNK通路的激活，而且还可抑制脑缺血诱导的NMDA受体NR2A亚基酪氨酸磷酸化；另外，一定浓度的乙醇也能激活GluR5抑制脑缺血诱导的JNK通路的活化。（3）脑缺血增强了兴奋性突触(E) NX-NLG1-PSD-95信号模块的组装，而抑制性突触(I) NX-NLG2信号模块组装无明显变化，结果提示:脑缺血E/I增加主要是由于兴奋性突触兴奋性增强引起的，预缺血通过NMDA受体可以抑制E/I的增加。
英文摘要
The mechanism about regulation of excitotary signaling regulation by inhibitory GABA signaling mechanism during brain ischemia insult is studied in this project, on rat cerebral ischemia model, focal cerebral ischemia and oxygen-glucose deprivation. The results from immunoblot, immunoprecipitation, electrophysiological recording and etc. show that: 1, co-application of agonists of GABA(A) receptor and GABA(B) receptor, muscimol and baclofen, not only inhibits the JNK activation through GluR6-PSD-95-MLK3 signaling module in the cerebral ischemia, but also inhibits the JNK activation through KA stimulation; and exerts neuroprotective effects through inhibiting NMDA receptor NR2A tyrosine phosphorylation, increasing nNOS phosphorylation, and inhibiting ASK1-JNK signaling by activating PI-3K/Akt signaling. 2, activation of GluR5-containing KA receptors has a neuroprotective effect against ischemic brain injury by suppressing NMDA receptor NR2A tyrosine phosphorylation and GluR6 mediated JNK activation; and a certain concentration of alcohol can activate GluR5 to inhibit JNK activaiton during ischemia. 3. ischemia leads to increased assembly of NX-NLG1-PSD-95 signaling module on excitotary synapses, whereas NX-NLG2 module on inhibitory synapses showes no significant change; the increased E/I ratio during ischemia i
结题摘要
本项目研究以全脑缺血或局灶脑缺血或缺糖缺氧（OGD）细胞模型，采用免疫沉淀，免疫印迹，细胞膜片钳技术等方法，研究了抑制性GABA信号对脑缺血诱导的兴奋性信号通路的调控机制。结果显示：（1）GABA(A)受体和GABA(B)受体激动剂联合用药，不但能抑制脑缺血诱导的GluR6介导的JNK通路的激活，而且还能抑制KA刺激引起JNK通路的激活；不但能抑制NMDA受体亚基NR2A酪氨酸磷酸化及增强nNOS的磷酸化，而且还可通过激活PI-3K/Akt通路抑制ASK1-JNK通路，从而达到保护神经元的作用。（2）GluR5的激动剂刺激突触前GluR5不但能抑制脑缺血诱导突触后GluR6介导的JNK通路的激活，而且还可抑制脑缺血诱导的NMDA受体NR2A亚基酪氨酸磷酸化；另外，一定浓度的乙醇也能激活GluR5抑制脑缺血诱导的JNK通路的活化。（3）脑缺血增强了兴奋性突触(E) NX-NLG1-PSD-95信号模块的组装，而抑制性突触(I) NX-NLG2信号模块组装无明显变化，结果提示:脑缺血E/I增加主要是由于兴奋性突触兴奋性增强引起的，预缺血通过NMDA受体可以抑制E/I的增加。