氧化应激/PARP-1介导自噬流受阻在妊娠期局麻药神经毒性中的作用

With the increasing in the birth rate and the high rate of cesarean section, as the main drug for cesarean anesthesia-local anesthetic related neurotoxicity complications have been widely concerned by scholars at home and abroad. While pregnancy can increase the neurotoxicity caused by local anesthetic, the mechanism is not clear. It is known that the level of oxidative stress in pregnant women is significantly higher during pregnancy.Previous studies have shown that oxidative stress may be an important mechanism, and oxidative stress can lead to DNA damage. Proteomics analysis showed that the injured DNA activated the repairase PARP-1 and autophagy-related pathways. Whether pregnancy increased the local anesthetic neurotoxicity related to the regulation of the pathway? Our pre-experimental found that after the neurotoxicity caused by local anesthetic during pregnancy, DNA damage repair enzyme PARP-1 was activated , While the cell autophagic flow was blocked; So we speculate: Pregnancy via oxidative stress / PARP-1 pathway leading to autophagic flow obstruction aggravate the neurotoxicity caused by local anesthetic. We propose to confirm the above hypothesis by immunohistochemistry, Western blotting, PARP-1 specific inhibitor/siRNA-PARP-1, autophagy in vivo model and so on in the model of neurotoxicity injury of local anesthetic in pregnant rats. This study provides new ideas and targets for further prevention and treatment of complications of high risk of neurotoxicity caused by cesarean anesthesia during pregnancy.

“二孩”政策使接受剖宫产的孕妇数量快速增长。临床研究表明：妊娠可加重局麻药导致神经毒性损伤，机制未明。已知妊娠期孕妇体内氧化应激水平明显增高，我们证实：氧化应激引发神经细胞DNA损伤与凋亡为局麻药致神经毒性损伤关键环节，同时DNA修复酶PARP-1及自噬相关通路参与此环节，妊娠期加重局麻药神经毒性损伤是否与上述通路调控失调有关？预实验发现：妊娠可加重局麻药致大鼠神经毒性损伤，DNA损伤修复酶PARP-1被过度激活，同时细胞自噬流受阻；由此推测：妊娠可通过氧化应激/PARP-1介导自噬流受阻加重局麻药所致神经毒性损伤。本项目拟建立妊娠大鼠局麻药神经毒性损伤模型，结合自噬在体检测，采用免疫组化、蛋白印迹、PARP-1特异性抑制剂/siRNA等方法证实上述假说，为妊娠期剖宫产局麻药引发神经毒性损伤的防治提供新思路和理论依据。

随着我国“二孩”政策的开放，接受剖宫产的孕妇数量急速上升。局部麻醉药是剖宫产麻醉的最主要用药，其正被广泛的用于产科麻醉与术后镇痛治疗。越来越多的临床和基础研究表明：妊娠可加重局麻药导致的外周神经损伤，其机制尚未阐明。本课题关注孕产妇及围生期安全，针对妊娠期加重局麻药导致的产妇神经毒性损伤这个亟待解决的科学问题，从妊娠期带来的产妇高氧化应激状态作为出发点展开研究。氧化应激一直被认为是导致围产期相关并发症的重要原因之一；我们的研究也发现：妊娠期产妇体内的氧化应激水平明显增高，同时产妇在接受局麻药后，其外周神经损伤明显加重。进一步，我们在妊娠小鼠的局麻药外周神经损伤模型中研究发现：妊娠可加重局麻药致小鼠神经毒性损伤，DNA损伤修复酶PARP-1被过度激活，导致细胞内自噬流受阻。使用PARP-1抑制剂PJ34可明显减轻由自噬流受阻导致的脊髓神经元的损伤；使用自噬抑制剂并不影响PARP-1的激活，但可部分逆转妊娠期局麻药导致的神经损伤。最后，我们的研究结果表明：通过氧化应激/PARP-1通路调控细胞自噬流受阻在妊娠加重局麻药神经毒性损伤过程中发挥重要作用，该研究可为防治妊娠期剖宫产麻醉高风险的神经毒性并发症提供新的思路和靶点。