NF-κB/Bcl-xL在氢气对蛛网膜下腔出血后早期脑保护中的作用机制研究

Subarachnoid hemorrhage (SAH) is a frequently seen devastating disease with high mortality and disability. Increasing evidence has proved that the early brain injury (EBI) period is closely bound up with the poor prognosis. However, the related mechanism remains elusive and there is no effective treatment. Recently, hydrogen (H2) has been a study focus after being approved as a selective anti-oxidative agent. However, related study mainly centered on the mechanism of selective anti-oxidative. In our previous study, we proved that H2 could alleviate EBI via reducing oxidative stress. Moreover, the first time, we found that H2 could activate NF-κB/Bcl-xL pathway after SAH. Hence, in order to verify the new mechanism, we will use RNAi in both vivo and vitro with the results of histology and neurological function combined to analyze the role of NF-κB/Bcl-xL pathway after H2 treatment in SAH model. New protective mechanism which can be the theoretical foundation of further study could be put forward from this experiment. Moreover, our study can provide important research basis for clinical translational research of H2.

蛛网膜下腔出血(Subarachnoid hemorrhage, SAH)后的早期脑损伤（early brain injury, EBI）是决定SAH 预后的重要因素,但尚无有效治疗方法。自氢气（Hydrogen，H2）被证明是能选择性清除毒性氧自由基而不影响正常氧化反应的治疗剂后，H2相关研究迅速成为热点，但仍主要围绕其减轻氧化应激机制展开。申请人在前期研究已证实H2可减轻SAH后脑氧化应激损伤的基础上，首次发现H2在SAH后可激活NF-κB/Bcl-xL抗凋亡通路。因此，本课题将围绕H2在SAH后能否通过NF-κB/Bcl-xL通路减轻EBI展开，拟通过体内、体外两个范畴的研究，在上下游分别应用RNA干扰技术抑制该通路, 结合神经损伤和神经功能评估结果验证这一新机制。通过本课题可提出H2对中枢神经保护作用的新机制，为后续研究提供新的理论基础，为H2治疗的临床转化研究提供前期基础。

蛛网膜下腔出血(subarachnoid hemorrhage, SAH)后的早期脑损伤（early brain injury, EBI）是决定SAH 预后的重要因素,但尚无有效治疗方法。自氢气（Hydrogen，H2）被证明是能选择性清除毒性氧自由基而不影响正常氧化反应的治疗剂后，H2相关研究迅速成为热点，但仍主要围绕其减轻氧化应激机制展开。申请人在前期研究已证实H2可减轻SAH后脑氧化应激损伤的基础上，首次发现H2在SAH后可激活NF-κB/Bcl-xL抗凋亡通路。因此，本课题围绕H2在SAH后能否通过NF-κB/Bcl-xL通路减轻EBI展开。通过体内、体外两个范畴的研究，应用RNA干扰技术抑制该通路, 结合神经损伤和神经功能评估结果验证这一新机制。研究结果显示H2治疗减轻了SAH后的脑损伤，改善了大鼠神经功能评分。通过对NF-κB/Bcl-xL进行检测，发现H2治疗提高NF-κB的蛋白表达，且激活NF-κB，同时明显提高Bcl-xL的基因转录及蛋白表达水平，再次验证我们前期的研究结果。在此基础上通过LV-ShRNA-Bcl-xl和NF-κB的抑制剂PDTC干预后发现H2对SAH后的神经保护作用被抑制。上述结果证明了我们前期推段H2在SAH后激活NF-κB/Bcl-xL抗凋亡通路起到神经保护作用的科学性。体外培养神经元建立的SAH模型也同样证明了H2对SAH后神经元的保护作用，且H2激活NF-κB/Bcl-xL通路。LV-ShRNA-Bcl-xl转染的神经元SAH后给予H2处理未能明显改善神经元损伤，因此体外神经元SAH模型同样证明了我们的推断。本研究为H2对中枢神经保护作用提供新的机制，为后续研究提供新的理论基础，为H2治疗的临床转化研究提供前期基础。