DR2/HSP90α/RIP3途径参与METH/高热诱导黑质神经元程序性坏死的机制

甲基苯丙胺（METH）神经毒性机理不清，长期困扰着临床和法医学界。申请人基于前一个NSFC项目资助，观察到METH与高热联合作用引起严重的黑质神经元坏死，这可能是METH损害大脑功能的重要途径，但具体机制不明。课题组前期研究结果显示，METH联合高热诱导HSP90α高表达，抑制HSP90α显著降低RIP3高表达并减少黑质神经元坏死，提示HSP90α调节RIP3机制在黑质神经元死亡中起重要作用；特异性阻断多巴胺受体2（DR2）降低HSP90α和RIP3高表达，提示DR2可能调控HSP90α/RIP3的分子变化。因此，项目拟采用基因干扰、GST pull-down、Co-IP、微透析和转棒实验等手段，从分子和动物水平解析DR2/HSP90α/RIP3调控轴参与METH/高热诱导黑质神经元坏死的机制，为METH相关脑损伤的法医学实践提供鉴定依据，为成瘾戒断、解毒靶点筛选提供新思路。

Lacking knowledge of methamphetamine (METH)-induced neurotoxicity is a question in forensic and clinical fields. Based on the previous NSFC project, we observed the combined effect of METH and hyperthermia that causes severe necrosis of neurons in the substantia nigra. This may be an essential way for METH to damage the brain in forensic and clinical practice, but the specific mechanism is not precise. We have previously confirmed that METH combined with hyperthermia induces increased expression of HSP90α, while inhibition of HSP90α significantly reduces necrosis of neurons and high expression of RIP3, suggesting that the regulation of RIP3 by HSP90α may play a key role in neuronal death; blocking dopamine receptor 2 (DR2) can reduce the level of HSP90α and RIP3, indicating that DR2 may be an upstream regulator of the HSP90α/RIP3 pathway. We would use genetic interference, GST pull-down, Co-IP, microdialysis, and animal behavior testing to verify the hypothesis “DR2 / HSP90α / RIP3 regulatory axis is involved in METH/hyperthermia-induced neuronal necroptosis in substantia nigra”, aiming to benefit forensic identification of METH related brain injury and to provide new ideas for addiction withdrawal and clinical intervention.