RIPK1/RIPK3/MLKL介导的程序性坏死参与七氟烷诱导的未成熟海马神经细胞毒性作用机制的研究

The adverse effects of anesthetic on the developing brain of infants have aroused wide concern. It has been confirmed that the sevoflurane, an inhalation anesthetic, causes cognition impairment via apoptosis of the immature hippocampal neurone. In our previous study, we found that sevoflurane exposure, with short time and low concentration, causes cognition impairment without apoptosis of the hippocampal neurone, which means that unknown mechanism must be involved. The latest research found that RIPK1/RIPK3/MLKL mediated necroptosis often co-exists with apoptosis and is also involved in the various neurotoxicity. Our preliminary experiment further found that the expression and binding force of necroptosis associated proteins in immature hippocampal neurons were significantly up-regulated after sevoflurane exposure. So we hypothesise that the RIPK1/RIPK3/MLKL mediated necroptosis is involved in sevoflurane induced neurotoxicity in immature hippocampal neuron. Based on our previous foundations, this project is aimed to verify our hypothesis through establishment cells and animal models of sevoflurane exposure, using MLKL gene knockout, RNA interference and animal behavior experiment. And we also explore strategies to prevent and improve the neurotoxicity of sevoflurane. Our aim is to provide a novel strategy for improving the safety of clinical anesthesis of the infants and children.

麻醉药物对婴幼儿未成熟大脑的毒性作用已引起广泛关注。现已证实，吸入性麻醉药物七氟烷通过诱导未成熟海马神经细胞凋亡，导致后期的认知功能障碍。申请者前期研究发现，低浓度短时间的七氟烷在没有造成海马神经细胞凋亡的情况下同样会导致后期认知功能障碍，提示有未知机制参与其中。最新研究表明，RIPK1/RIPK3/MLKL介导的程序性坏死常与凋亡共存，并参与了多种神经细胞的毒性作用。申请者预实验进一步发现七氟烷可显著上调未成熟海马神经细胞中程序性坏死相关蛋白的表达量和结合力。据此提出假说：程序性坏死可能参与了七氟烷对未成熟海马神经毒性作用的机制。本课题拟在前期基础上，通过建立七氟烷暴露细胞及动物模型，利用MLKL基因敲除、RNA干扰及动物行为学实验等技术，明确程序性坏死参与七氟烷诱导的未成熟海马神经毒性作用的机制，并探索预防和改善七氟烷神经毒性作用的策略。旨在为提高临床婴幼儿麻醉安全性提供新的理论依据。

麻醉药物对婴幼儿未成熟大脑的毒性作用的机制已经在全世界深有研究。现已证实，吸入性麻醉药物七氟烷通过各种机制会导致后期的认知功能障碍。申请者前期研究发现，低浓度短时间的七氟烷在没有造成海马神经细胞凋亡的情况下同样会导致后期认知功能障碍，提示有未知机制参与其中。通过本课题的资助，申请者证实了七氟烷可显著上调未成熟海马神经细胞中程序性坏死相关蛋白的表达量和结合力。并且在本课题的资助下通过建立七氟烷暴露细胞及动物模型，利用MLKL基因敲除、RNA干扰及动物行为学实验等技术证实了程序性坏死参与了七氟烷对未成熟海马神经毒性作用机制的假说。明确了程序性坏死参与七氟烷诱导的未成熟海马神经毒性作用的机制，并探索预防和改善七氟烷神经毒性作用的策略。旨在为提高临床婴幼儿麻醉安全性提供新的理论依据。