MicroRNA相关信号通路轴在颅脑创伤昏迷及电刺激干预中的作用机制

Traumatic coma remains the complex complication after severe head trauma and brings huge health care burdens. The proposed grant focused on the scientific hypothesis that microRNAs and related pathways involving in the traumatic response in reticular formation of brain stem after mechanical insults. The research works will include determination of excitatory and inhibitory synaptic transmitters related with the SNARE, JAK-STAT pathway in reticular formation in midbrain, setting up mir-1275-vamp2-snap25 transfect cell strain to identify the transmitter releasing effect and constructing PC12-mir-93 cell strain to explore the regulatory effect of mir-93 on Jak-Stat pathway, so that to confirm the in vitro effect of microRNA Related pathways. Both the regulation and function of pathways will be identified in vivo and the researcher will apply right median nerve electrical stimulation as a intervene technique to explore the therapeutic mechanism underlying brain stem functional recovery after head injury. Through the above lab workings, the proposed grant will clarify that the microRNAs and related pathways are biomarkers and treating targets in reticular formation after traumatic coma, and disclose the protective effect of right median nerve stimulation on unconsciousness following insult of brain trauma.

颅脑创伤昏迷严重影响患者预后，其发生机制和治疗手段有待深入探索。本课题基于前期研究结果，拟针对微RNA参与脑干网状结构创伤应答调控这一假说开展研究，观察微RNA相关SNARE、JAK-STAT信号通路中兴奋性/抑制性神经递质Syt2/vGlut1在脑外伤后中脑网状结构的表达差异，对mir-1275-VAMP2-SNAP25和mir93-IFNK-JAK-STAT信号通路通过构建PC12稳转细胞系进行神经递质释放等体外生物效应验证，利用大鼠实验性颅脑创伤模型对上述信号通路物质在创伤昏迷后调控变化进行体内验证，采用经皮右正中神经电刺激干预技术对创伤昏迷恢复机制进行观察。通过上述研究阐明微RNA在颅脑创伤所致昏迷中具有生物标志物、生物反应关键环节和治疗靶标的多重生物学意义，证实微RNA及其相关信号通路是颅脑创伤昏迷相关研究的重要节点，探索促醒措施的唤醒机制，推进意识障碍及促醒治疗的研究进步。

创伤性脑外伤的医治是我国待解决的重要医学问题，研究创伤性脑外伤的病理机制和开发有效的检测标志物是解决该问题的重要方向。在过去的四年中，课题组分别从上述两个方向展开研究，在miR-1275-SOX7通路和泪液标志物方面取得一定成果。首先，我们通过细胞和小鼠模型发现，miR-1275在创伤后表达上调，促进神经损伤。miR-1275在该过程中的主要靶点为SOX7，通过细胞和小鼠模型发现SOX7具有创伤后神经保护作用，SOX7的过表达可削弱miR-1275的神经损伤作用，从而提供了通过抑制miR-1275靶向治疗创伤性脑外伤的治疗思路。因此，课题发现miR-1275-SOX7通路是创伤性脑外伤后的重要病理机制。其次，课题通过对颅脑创伤后植物状态患者泪液的蛋白质组学研究发现7个泪液蛋白标志物（CTSB, PRSS1, S100A7, GSTP1, CFH, KNG1, ORM1），可有效的鉴别脑创伤后的病理状态，并为研究创伤性脑外伤的系统性病理机制提供科学基础。